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Cambridge Healthtech Institute’s 14th Annual JANUARY 19-23, 2015

Town and Country Resort & Convention Center

SAN DIEGO, CA

Cambridge Healthtech Institute, 250 First Avenue, Suite 300 , Needham, Massachusetts 02494Telephone: 781-972-5400 • Toll-free in the U.S. 888-999-6288 • Fax: 781-972-5425

PREMIER SPONSORS:

2015 Event Features:

• 1,200+ International Participants including Scientists, Regulators and Solution Providers

• 20 Conferences on Antibodies, Formulation, Expression, Analytics, Purification and more

• 13 Short Courses to Enhance Your Learning Experience

• 325+ Scientific Presentations from Industry Leaders

• 80+ Interactive BuzZ Session Roundtables

• 100+ Exhibitors Showcasing Novel Technologies and Solutions

• 125+ Cutting-Edge Research Posters

PLENARY KEYNOTE

From Yeast to the Brain: Advances in Proteomics

John R. Yates, Ph.D., Ernest W. Hahn Professor, Chemical Physiology and Molecular and Cellular Neurobiology, The Scripps Research Institute

Sponsorship & Exhibit Opportunities

PROTEIN ENGINEERING & DEVELOPMENT

Recombinanat Protein Therapeutics

Enhancing Antibody Binding & Specificity

Improving the Clinical Efficacy of Antibody Therapeutics

ANTIBODY THERAPEUTICS

Cancer Targets for Antibody Therapeutics

Antibody-Drug Conjugates

Bispecific Antibody Therapeutics

FORMULATION & STABILITY

Optimizing Biologics Formulation Development

Lyophilization & Emerging Drying Technologies

Protein Aggregation & Emerging Analytical Tools

EXPRESSION & PRODUCTION

Engineering Genes, Vectors, Constructs & Clones

Recombinant Protein Expression & Production

Transient Protein Production

ANALYTICS & IMPURITIESCharacterization of ADCs, Bispecifics & New Biotherapeutics

Detection and Characterization of Particulates & Impurities

Extractables & Leachables

PROCESS TECHNOLOGIES & PURIFICATION

Single-Use Technologies & Continuous Processing

Protein Purification & Recovery

Higher-Throughput Protein Purification

ACCOMPANYING CONFERENCES:• MEMBRANE PROTEINS• CHO CELLS

Hotel & Travel / Additional Info

Registration & Pricing

Register online at CHI-PepTalk.com

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Register by September 12 for Early-Bird

Savings up to $600

https://chidb.com/register/2015/ptk/reg.asp

PepTalk: The Protein Science Week is one of the largest gatherings of protein science researchers in the United States, and when you bring together some of the most influential people in the field - big things happen!

PepTalk offers an array of education, innovation and networking programs. Over 300 high-caliber speakers share case studies, unpublished data, breakthroughs and solutions that support and enhance your research. Ample networking opportunities allow you to connect with colleagues and

peers from around the world and gain new perspectives on the evolution of biologics. Choose between 20 Conferences, 13 Short Courses, 3 Training Seminars, 80+ BuzZ Session Discussion Roundtables, and dedicated exhibit hall and poster viewing hours to create a custom agenda that fits your research and networking needs!

PLENARY KEYNOTE Wednesday, Jan. 21, 4:30pm

4:30 Plenary Keynote Introduction

4:40 From Yeast to the Brain: Advances in ProteomicsJohn R. Yates, Ph.D., Ernest W. Hahn Professor, Chemical Physiology and Molecular and Cellular Neurobiology, The Scripps Research Institute

5:25-5:30 First Annual PepTalk STAR AwardABOUT:John R. Yates is the Ernest W. Hahn Professor in the Department of Chemical Physiology and Molecular and Cellular Neurobiology at The Scripps Research Institute. His research interests include development of integrated methods for tandem mass spectrometry analysis of protein mixtures, bioinformatics using mass spectrometry data and biological studies involving proteomics. He is the lead inventor of the SEQUEST software for correlating tandem mass spectrometry data to sequences in the database and developer of the shotgun proteomics technique for the analysis of protein mixtures. His laboratory has developed the use of proteomic techniques to analyze protein complexes, posttranslational modifications, organelles and quantitative analysis of protein expression for the discovery of new biology. Many proteomic approaches developed by Yates have become a national and international resource to many investigators in the scientific community. He has received the American Society for Mass Spectrometry research award, the Pehr Edman Award in Protein Chemistry, the American Society for Mass Spectrometry Biemann Medal, the HUPO Distinguished Achievement Award in Proteomics, Herbert Sober Award from the ASBMB and the Christian Anfinsen Award from The Protein Society. He was ranked by Citation Impact, Science Watch as one of the Top 100 Chemists for the decade, 2000-2010. He was #1 on a List of Most Influential in Analytical Chemistry compiled by The Analytical Scientist 10/30/2013 and is on the List of Most Highly Influential Biomedical Researchers, 1996-2011, European J. Clinical Investigation 2013, 43, 1339-1365.He has published 751 scientific articles with ~57,000 citations, and an H index 119.

ALL PARTICIPANTS ARE WELCOME!

Get Connected!

PRE-CONFERENCE DINNER SHORT COURSES*

Sunday, Jan. 18

Monday-Tuesday, Jan. 19-20

DINNER SHORT COURSES* Tuesday, Jan. 20

Wednesday-Thursday (am), Jan. 21-22

Thursday (pm)-Friday, Jan. 22-23

PIPELINE 1 Protein Engineering &

DevelopmentShort Courses Recombinant Protein Therapeutics Short Courses Enhancing Antibody Binding and

SpecificityImproving the Clinical Efficacy of

Antibody Therapeutics

PIPELINE 2Antibody Therapeutics Short Courses Cancer Targets for Antibody

Therapeutics Short Courses Antibody-Drug Conjugates Bispecific Antibody Therapeutics

PIPELINE 3Formulation & Stability Short Courses Optimizing Biologics Formulation

Development Short Courses Lyophilization and Emerging Drying Technologies

Protein Aggregation and Emerging Analytical Tools

PIPELINE 4Expression & Production Short Courses Engineering Genes, Vectors,

Constructs and Clones Short Courses Recombinant Protein Expression and Production Transient Protein Production

PIPELINE 5Analytics & Impurities Short Courses

Characterization of ADCs, Bispecifics and New

BiotherapeuticsShort Courses Detection and Characterization of

Particulates and Impurities Extractables and Leachables

PIPELINE 6Process Technologies &

PurificationShort Courses Single-Use Technologies and

Continuous Processing Short Courses Protein Purification and Recovery Higher-Throughput Protein Purification

NEW Accompanying Conferences Short Courses Short Courses Membrane Proteins / CHO Cells

Training Seminars Short Courses Intro to Bioprocessing Short Courses

Intro to Formulation

Intro to Analytical Method Development and Validation

for Therapeutic Proteins

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CORPORATE SPONSORS

PREMIER SPONSORS

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SC1: Production Challenges for Complex Biologics: ADCs, Bispecifics and Fusion ProteinsThis course addresses the typical production issues encountered with complex biologics, namely fusion proteins, antibody-drug conjugates and bispecific antibodies. Experts elucidate the structure and nature of these biologics in order to understand and master their properties. Along with exploring manufacturing challenges, the course also reveals how to overcome these challenges with practical insights and advice.Instructors: Stefan Schmidt, Ph.D., Vice President, DSP, Rentschler BiotechnologyChristopher D. Thanos, Ph.D., Director, New Molecular Entities, Halozyme Therapeutics, Inc.George Octavian Badescu, Ph.D., Director, Scientific Affairs, Bioconjugation & Protein Engineering, PolyTherics Ltd.

SC3: A Rational Approach to Formulation Development of Biologic TherapeuticsThe course offers a forum discussing how to develop formulation for biologic drugs. Case studies demonstrate how to incorporate Quality-by-Design (QbD) concepts to design multivariate experiments, how to obtain representative data and how to analyze data in order to propose sound formulation of drug substance or drug product in the context of designated container closure systems. The course will combine how-to suggestions and real-world examples in an interactive discussion.Instructors: Kevin Zen, Ph.D., Senior Manager, Biologics Development, AllerganSteven LaBrenz, Ph.D., Scientific Director, Drug Product Development, Janssen R&D

SC4: Genome Editing Using CRISPRMammalian cells are the workhorses for biopharmaceutical production. Thus, genome engineering/editing of these hosts to improve product quality and yields are of great interest. CRISPR, the newest gene editing tool, is gaining popularity among protein engineers and cell line developers. This course provides an introduction to CRISPR technology and insights on implementation for your protein expression and production pipeline.Instructors: Helene Faustrup Kildegaard, Ph.D., Co-Principal Investigator, Novo Nordisk Foundation Center for Biosustainability, Technical University of DenmarkNorman Garceau, Ph.D., CSO, Blue Sky Biotech, Inc.Deepak Reyon, Ph.D., Scientist I, Genetically Engineered Model Center, Biogen Idec

SC5: Accelerated Stability Testing of BiologicsThis short course guides the researcher in designing studies for accelerated stability testing of biologics. The course begins with basic underlying concepts governing protein drug product stability, and focuses on design principles for measuring stress and accelerated stability testing of not only the protein of interest, but also excipients and primary packaging components. Strategies to handle complexities arising from their interactions will also be discussed.Instructors:Yatin R. Gokarn, Ph.D., Drug Product and Device, Biologics Development, Gilead Sciences, Inc. Vishal C. Nashine, Ph.D., Senior Research Investigator, Drug Product Science & Technology, Bristol-Myers Squibb Co.

SC6: Establishing the Business Case for Single-Use and Continuous ProcessingThis short course introduces attendees to the paradigm shift and a new way of economic and manufacturing considerations for implementing single-use systems and continuous processing. Based on case studies, projects and available data, we establish a platform for drug manufacturing that is robust, streamlined, sustainable and energy saving, and at the same time reduces COGS and carbon print, culminating towards a more streamlined operation for either batch or continuous processing.

Instructor: Robert Dream, PE, CPIP, CPMP, Ph.D., Principal, HDR Company Ltd.

PRE-CONFERENCE DINNER SHORT COURSES*

BuzZ Sessions are facilitated, small-group discussions. Interactive participation leads to problem-solving solutions and future collaborations around focused topics.

If you have a topic idea or would like to moderate a table, please contact: Ann Nguyen at [email protected]

Please visit our website for more details.

SUNDAY, JANUARY 18 | 5:00-8:00 PM

* Please visit our website for more details. Separate registration required.





























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SC7: Targeting of GPCRs with Monoclonal AntibodiesWhile GPCRs (G protein-coupled receptors) are important therapeutic targets, it has been challenging to discover therapeutically relevant antibodies against them. This course examines different steps along the anti-GPCR antibody discovery pathway and highlights various approaches to accomplishing each step. Topics include: 1) Antibody discovery, 2) Assays to measure antibody binding, 3) In vitro assays to measure functional activity of the antibody, and 4) Review of promising GPCR targets and antibodies in the clinic.Instructor: Barbara Swanson, Ph.D., Director, Research, Sorrento Therapeutics, Inc.

SC8: Affecting Effector Function: Engineering the Fc RegionThere are a growing number of antibodies and Fc fusion proteins in development that contain a modified Fc region, either via changes in amino acid sequence or in glycoforms. Engineering antibodies and Fc fusion proteins has become more sophisticated at generating molecules that are better suited to the pharmacological activity required. This course focuses on characterizing and engineering effector functions in order to create more effective therapeutics.Instructors: Steven Chamow, Ph.D. Principal Consultant, Chamow & Associates, Inc. Tomoyuki Igawa, Ph.D., Manager, Antibody Engineering Group, Discovery Research, Chugai Pharmaceutical Co., Ltd.Futa Mimoto, Ph.D., Researcher, R&D, Chugai Pharmaceutical Co., Ltd.Tilman Schlothauer, Ph.D., Senior Scientist, Protein Analytics, Roche Diagnostics GmbH

SC9: Protein Aggregation: Mechanism, Characterization and ConsequencesProtein aggregation is recognized by regulatory agencies and the biopharmaceutical industry as a key quality attribute of biotherapeutic products. Various aggregates hold the potential for adversely impacting production and patients in a variety of ways. This in-depth workshop reviews the origins and consequences of aggregation in biotherapeutics, and then examines strategies for predicting and quantifying aggregation in biopharmaceuticals. It benefits scientists engaged in development, production, analytical characterization and approval of biotherapeutics and who require a good working knowledge of protein aggregation.Instructors: Thomas Laue, Ph.D., Professor, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New HampshireDavid F. Nicoli, Ph.D., Vice President R&D, Particle Sizing Systems, LLC

SC10: Transient Protein Production in Mammalian CellsThis short course introduces both the fundamental concepts and technologies needed to establish transient protein production in mammalian cells. This allows for the rapid generation, purification and characterization of milligram-to-gram quantities of secreted or intracellular recombinant proteins for therapeutic, functional and structural studies. The course combines instruction and case studies in an interactive environment.Instructors: Richard Altman, MS, Research Scientist, Molecular Sciences, Alexion PharmaceuticalsHenry C. Chiou, Ph.D., Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher ScientificDominic Esposito, Ph.D., Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc.

SC11: Materials in Contact with Biologics: Understanding Risk to Quality and SafetyMaterials that contact biologics during manufacturing, storage and final packaging can pose risks to biopharmaceutical quality. This in-depth course reviews the regulatory requirements, types of materials used and material chemistry, and then examines the strategies for prediction and risk assessment of potential threats to quality and safety of biologics drug products. The course reviews development of a successful analytical strategy for single-use components, container closure components and risk posed by leachables.Instructors: Jeffrey Carter, Ph.D., Strategic Projects Leader, GE Healthcare Diane Paskiet, Ph.D., Director, Scientific Affairs, West Pharmaceutical

SC12: Protein Purification Strategies: Dealing with Proteins that Are Prone to AggregateThis course provides a comprehensive and detailed outline of hands-on issues for purifying proteins. We first address general considerations about the protein we want to produce, including issues of activity, solubility, homogeneity, purity and proper oligomeric conformation. Aggregation is one of the main obstacles in protein production, so we look at how to monitor for aggregation and comprehend its mechanism. We also discuss how to check for the optimal solubility conditions at the expression level, and our comprehensive approach for optimizing solubility during purification. We also discuss expression screening methodology, environmental factors to consider during purification, families of additives and screening for additives. Lastly, we address ways to avoid aggregation, as well as setting up protein concentration and storage.Instructor: Mario Lebendiker, Ph.D., Head, Protein Purification Facility, Wolfson Centre for Applied Structural Biology, Hebrew University of Jerusalem

* Please visit our website for more details. Separate registration required.

DINNER SHORT COURSES* TUESDAY, JANUARY 20 | 5:00-8:00 PM





























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TS1: INTRODUCTION TO BIOPROCESSING Instructors:

Susan Dana Jones, Ph.D., Vice President and Senior Consultant, BioProcess Technology Consultants, Inc.

Sheila G. Magil, Ph.D., Senior Consultant, BioProcess Technology Consultants, Inc.

CHI’s Introduction to Bioprocessing training seminar offers a comprehensive survey of the steps needed to produce today’s complex biopharmaceuticals from early development through commercial. The seminar begins with a brief introduction to biologic drugs and the aspects of protein science that drive the intricate progression of analytical and process steps that follows. We then step through the stages of bioprocessing, beginning with the development of cell lines and ending at the packaging of a finished drug product. The seminar also will explore emerging process technologies, facility design considerations and the regulatory and quality standards that govern our industry throughout development. The important roles played by the analytical and formulation steps in developing and gaining approval for a biopharmaceutical are also examined.

This 1.5-day class is directed to attendees working in any aspect of industry, including scientific, technical, business, marketing or support functions, who would benefit from receiving a detailed overview of this field.

About the Instructors:Susan Dana Jones is a seasoned biotechnology entrepreneur with experience in product development, outsourcing and strategic planning. Dr. Jones is a subject matter expert in cell line development and characterization for biosimilar, new biopharmaceutical, and vaccine development programs. She has broad knowledge of regulatory requirements for manufacturing products for human use and has prepared CMC sections of multiple regulatory submissions. She currently serves on the Board of Directors of Gene Solutions, the Scientific Advisory Board of Symphogen, and is a member of the Editorial Advisory Board of BioProcess International. She received her Ph.D. in Genetics from the University of California, San Francisco.Sheila Magil has over 20 years of experience in quality and analytical method development for biologics, peptides and small molecules. Her expertise includes quality assurance, protein and peptide biochemistry, and analytical development. She was formerly Senior Manager of Analytical Development and Quality Control at Biomeasure, Inc., and previously held positions at WaratahPharma, Alkermes, Bion, and HHMI at Massachusetts General Hospital. Dr. Magil has implemented quality systems and has managed external analytical and QC activities for multiple biopharmaceutical products. Dr. Magil holds a Ph.D. in Biochemistry from the University of Minnesota.

TS2: INTRODUCTION TO BIOLOGICS FORMULATION AND DELIVERY

Instructor: Timothy Kelly, Ph.D., Vice President, Biopharmaceutical Development, KBI Biopharma, Inc.

Pooja Arora, Ph.D., Associate Director, Biotherapeutics Global CMC Regulatory Affairs, PfizerThe course focuses on strategies to plan and execute preformulation and formulation development

studies for biologics, which require co-optimization of multiple physical, chemical and conformational stability attributes while operating under accelerated timelines to deliver the drug to the clinic. The course begins with an overview of biophysical and biochemical properties of proteins. A typical development workflow (including statistical analysis and DOE elements) will be outlined to demonstrate the core elements employed during protein formulation. The course concludes with real-world examples from formulation development projects for liquid and lyophilized products.

• Basics of protein biochemistry, with focus on folding mechanism, stability and structural hierarchy

• Degradation pathways relevant to biologics shelf life

• Biophysical and analytical characterization tools

• Typical workflow for biologics formulation development projects

• Introduction to common delivery devices

About the Instructors:Tim Kelly has over 20 years of experience in protein and nucleic acid characterization. In his role at KBI Biopharma, Tim is responsible for analytical development, formulation development, and quality control. Prior to joining KBI Biopharma, Tim held the position of Director of Quality Control for Diosynth Biotechnology, where he was responsible for method validation, in-process control, release and stability of clinical and commercial biopharmaceutical products. Tim’s experience also includes the analytical development, formulation development, characterization and/or production of more than 200 clinical and commercial protein therapeutics, including monoclonal antibodies, enzymes, cytokines, fusion proteins, PEGylated proteins, protein vaccines and peptides. Tim has led the successful formulation development of over 95 clinical and commercial biopharmaceutical products, including liquid and lyophilized dosage forms for intravenous and subcutaneous administration, at protein concentrations ranging from 10µg/mL to 200mg/mL. Tim earned his Ph.D. in Molecular Genetics & Biochemistry from Georgia State University.

Pooja Arora is an Associate Director in the Biotherapeutics Global CMC – Regulatory Affairs Department at Pfizer. Pooja has more than thirteen years of experience in protein biophysical and analytical characterization. Prior to joining Pfizer, Pooja has held positions in Genentech-Roche, Bristol Myers Squibb and KBI Biopharma. Her responsibilities in prior roles have included development of robust drug product and manufacturing process for ready-to-use solution and lyophilized protein therapeutics in vials and pre-filled syringes. She has led teams responsible for scale-up and technical transfer of manufacturing process for drug product to both clinical and commercial manufacturing sites. Pooja earned her M. Sc. in Chemistry from Indian Institute of Technology (Delhi) and Ph.D. in Chemistry from Duke University.

TS3: INTRODUCTION TO ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR THERAPEUTIC PROTEINS

Instructor: Jichao (Jay) Kang, Ph.D., RAC, Director, Analytical and Formulation Development, Gallus Biopharmaceuticals NJ, LLCThis course is a panoramic review of analytical

method development and validation for therapeutic proteins, including antibodies and enzymes. It is intended for scientists working on therapeutic proteins in Analytical Development, Quality Control, Product Development or related functional areas. It starts with basic knowledge of work on therapeutic proteins: manufacturing of proteins drugs, regulatory affair knowledge and protein chemistry. It then discusses fundamentals and practical aspects of commonly used analytical methods for proteins, including methods for structure elucidation, glycan characterization, biophysical characterization, potency measurement, purity and impurity analysis. The course concludes with the strategy and common practice in method validation and method transfer, including regulatory compliance at different stages of product development, application of DOE and QbD. The course emphasizes practical applications, real-world examples and useful tips.

Benefits

• Gain a complete picture of analytical method development and validation process

• Gain a basic understanding of commonly used analytical methods for proteins

Who Should Attend

• Analytical development scientists, process development scientists, QC analysts, regulatory affair managers, project managers and quality assurance managers

About the Instructor:Dr. Jichao Kang holds a Ph.D. in Pharmaceutics and has been working on characterization, method development and validation and formulation for protein therapeutics since 1995. He is an accomplished researcher with over 15 peer-reviewed journal articles and book chapters, several patents and numerous conference presentations. The proteins he has worked on extensively include cytokines, antibodies, enzymes and protein conjugates. He is a key contributor in dozens of IND/IMPD and BLA/MAA filings. He is currently the Director of Analytical and Formulation Development at Gallus BioPharmaceuticals NJ, LLC, one of the leading CMOs for biologics, and held the same position at Laureate BioPharma before it was acquired by Gallus. Prior to Laureate, he was the department head of Analytical Development at Auxilium Pharmaceuticals, Inc., and was a key contributor in Auxilium’s successful marketing application of Xiaflex in both U.S. and EU. He also worked in MedImmune, PDL, and Neose Technologies.

JANUARY 19-20, 2015 DAY 1 8:30 AM - 5:30 PM | DAY 2 8:30 AM - 12:30 PM

JANUARY 21-22, 2015 DAY 1 8:30 AM - 4:25 PM | DAY 2 8:30 AM - 12:30 PM

Please visit our website for more details.

Cambridge Healthtech

Each CHI Training Seminar offers 1.5 Days of instruction with start and stop times for each day shown above and on the Event-at-a-Glance published in the onsite Program & Event Guide. Training Seminars will include morning and afternoon refreshment breaks, as applicable, and lunch is “on your own”.

Each person registered specifically for the training seminar will be provided with a hard copy handbook for the seminar in which they are registered. A limited number of additional handbooks will be available for other delegates who wish to attend the seminar, but after these have been distributed no additional books will be available.

Those not attending a full training seminar, but who are registered for other tracks on those days of the conference, may participate in the training seminar sessions. We ask that those joining the seminars while they are in progress enter the room only during scheduled break periods to avoid disrupting the class in progress.





























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PIPELINE 1: PROTEIN ENGINEERING & DEVELOPMENT

11th Annual Recombinant Protein Therapeutics

Fusion Proteins and Beyond

JANUARY 19-20





























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The customizable functionality of fusion protein therapeutics creates advantages over antibody-based therapies by combining modular building blocks that can reach targets not accessible to antibodies. Additional advantages include lower patient dosing, reduced production costs and improved product homogeneity. The Recombinant Protein Therapeutics meeting explores the varying constructs and “designs” of fusion protein molecules, and discloses how they are being engineered to form more efficacious therapeutics that offer specificity with enhanced stability and longer half life. Experts present case studies from R&D through clinical data and share the results they have achieved.

SUNDAY, JANUARY 18

4:00-5:00 pm Short Course Registration

5:00-8:00 Pre-Conference Dinner Short Courses See pages 4-5 for details

4:00-8:00 Main Conference Registration

MONDAY, JANUARY 19

7:30 am Conference Registration and Morning Coffee

NEXT-GENERATION BIOLOGICS

9:00 Chairperson’s Opening RemarksStefan Schmidt, Ph.D., Vice President, DSP, Rentschler Biotechnology

»KEYNOTE PRESENTATION

9:10 Roche’s Strategies to Discover, Design, Develop, and Deliver New Innovative Therapeutic BiologicsRalf Schumacher, Ph.D., Site Head, Large Molecule Research Penzberg, and pRED Center Manager, Roche Diagnostics GmbHBiologics have become a key component in the treatment of various life-threatening diseases. The majority of these drugs are classical monoclonal antibodies. In order to discover and develop differentiated monoclonal antibodies, Roche’s strategy is based on engineering technologies. ADCC-enhancement, multi-pathway-inhibition, specific tumor-targeting of pharmacophores and blood-brain-barrier crossing are examples for successfully engineered mAbs and fusion proteins. In this presentation, I will describe Roche’s strategies to design such molecules, give examples but will also address challenges for technical development.

9:30 Ubx as a Novel Protein-Based Material: Structural Insights, Functionalization via Protein Fusion, and Biomedical ApplicationsShang-pu Tsai, Ph.D. Candidate, Molecular and Cellular Medicine, Texas A&M University

»FEATURED PRESENTATION

9:50 Monomeric Fc Fusion Clotting Factors for the Treatment of HemophiliaJennifer Dumont, Ph.D., Director, Medical Affairs, Biogen Idec, Inc.Prophylactic clotting factor replacement in patients with hemophilia improves outcomes, but requires frequent injections with traditional therapies. Recombinant fusion of the Fc domain of human IgG with clotting factors VIII and IX was performed to extend the half-life of these factors and provide the potential to reduce the frequency of injections needed to control bleeding in hemophilia A and B patients, respectively. The Fc fusion factors are configured with monomeric factor VIII or IX covalently linked to dimeric Fc. These protein fusion products have recently been approved for the treatment of hemophilia and the development programs will be discussed.

10:20 Coffee Break

ENGINEERING BREAKTHROUGHS

10:45 A Small Biologic Alternative to PCSK9 Antibodies: Pharmacologic Profile and Demonstration of Robust LDL Lowering with an Anti-PCSK9 AdnectinTracy Mitchell, Ph.D., Principal Scientist, Bristol Myers-Squibb Co.PCSK9 is perhaps the most promising drug target for treating cardiovascular disease since the discovery of statins. Compared with therapeutic IgG antibodies currently in clinical trials, targeting circulating PCSK9 with a smaller molecular scaffold could offer reduced dose burdens and different pharmacologic profiles. We present the pharmacological profile of BMS-962476, a potent Adnectin inhibitor of PCSK9 that has demonstrated robust target engagement and LDL lowering in mice, cynos and humans.

11:15 Development of an Intein-Based Conjugation Platform for the Synthesis of Fc-Fusion ProteinsOliver Thiel, Ph.D., Principal Scientist, Chemical Process Research & Development, Amgen, Inc.Identification of an ideal platform technology for conjugation of small molecules and peptides to biomolecules for improved pharmacokinetics has been a recent focus within academic and industrial laboratories. This contribution will focus on the development of an intein-based platform for conjugation of peptides at the C-terminus of the Fc domain of immunoglobulins. In the course of platform development, selection of intein, cleavage residue, and linker between Fc and intein were examined.






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11:45 A Bi-Functional Antibody-Receptor Domain Fusion Protein Simultaneously Targeting IGF-IR and VEGF for DegradationYang Shen, Ph.D., Research Advisor, Antibody Engineering, Eli Lilly & CompanyA fully human Bi-functional Antibody-receptor domain (VEGFR1 domain 2) fusion molecule with ligand Capture (BiAbCap) targeting IGF-IR and VEGF was designed and developed, that displays excellent thermal and physical stability. Taking advantage of natural receptor-ligand interaction, bi-AbCap represents a novel and developable format of bi-functional antibodies with potent neutralizing activities against both targets. The unique “capture-for-degradation” mechanism translates to potent anti-tumor activity superior to the combination in vivo.

12:15 pm Reinventing Immunoassays: Introducing the SimplePlex

Sponsored by

Rajiv Pande, Ph.D., Vice President, ProteinSimpleAlthough ELISA is routinely used for measuring individual protein analytes it has limitations in translational research applications where precise and accurate analysis of multiple analytes from small sample volumes is required. ProteinSimple has developed the Simple Plex, a fully automated, hands-free immunoassay platform that retains the specificity of single-analyte ELISAs, but provides rapid multi-analyte analysis. The Simple Plex integrates a desktop analyzer with a disposable microfluidic cartridge to deliver superior performance and unprecedented ease of use in research and diagnostic applications.

12:45 Session Break

1:00 Luncheon Presentation: Applications of Acoustic Membrane Micro Particle in Large Molecule Bioanalysis

Sponsored by

Johanna Mora, Ph.D., Senior Research Investigator II, Bristol-Myers SquibbIn cases where established technologies cannot deliver on the assay sensitivity requirements set by a specific drug development program, alternative platforms may need to be evaluated. This presentation will share a case study on the application of the Acoustic Membrane Micro Particle (AMMP) technology for PK determination of a domain antibody and cover results on accuracy and precision and selectivity assessments. The second case study will cover the development of an immunogenicity assay on the AMMP where none of the established platforms were able to achieve adequate sensitivity.

ENHANCING PROPERTIES

2:00 Chairperson’s RemarksManfred Schuster, Ph.D., COO, Apeiron Biologics AG

2:05 Unravelling the Molecular Basis of Host Cell-Specific Glycosylation and Species-Specific PharmacokineticsCatherine Huntington, Ph.D., Research Scientist II, Antibody Discovery and Protein Engineering (ADPE), MedImmune, LLCGlycosylation plays a significant role in the half-life of recombinant proteins in vivo, with the production cell lines affecting PK. Here, we expand on observations that HEK-293 expressed proteins are rapidly cleared in mouse models and our efforts to characterise the glycan forms responsible. Additionally, we will present our work to develop methods to profile glycan forms on recombinant proteins with the aim to predict impact on half-life of different species.

2:35 Enhancing Stability of the Therapeutic Enzyme L-Asparaginase by Biocompatible NanoparticlesManfred Konrad, Ph.D., Research Director, Enzyme Biochemistry, Max Planck Institute for Biophysical ChemistryThe enzyme L-asparaginase is a protein drug of high value in antileukemic therapy. Clinically approved L-asparaginases are of bacterial origin, though they elicit severe side effects, in particular immunogenicity. We present a novel approach for encapsulation of the enzyme using calcium carbonate particles surrounded by layers of biocompatible polymers, thus forming nanocontainers as carriers to enhance serum stability and suppress recognition by the immune system.

3:05 Manufacturing of Half-Life Extended Fusion Proteins: Current Trends and ChallengesStefan Schmidt, Ph.D., Vice President, DSP, Rentschler BiotechnologySecond and third generation therapeutic proteins often contain a half-life extension moiety. These additional modules of fusion proteins often complicate the manufacturing process as they require specific adaptations of both up- and downstream processes. First, we give a comprehensive overview on the current state-of-the-art regarding half-life extension technologies, and second, we illustrate manufacturing challenges and solutions presented through selected case studies, additionally giving practical advice on optimization potential.

3:35 Sponsored Presentation (Opportunity Available)

3:50 Refreshment Break

CONQUERING DISEASE

4:15 A Neuroprotective Brain-Penetrating Endopeptidase Fusion Protein Ameliorates Alzheimer Disease Pathology and Restores NeurogenesisEliezer Masliah, M.D., Professor, Neuroscience and Pathology, University of California at San DiegoAlzheimer’s (AD) and Parkinson’s Disease (PD) are the most common neurodegenerative disorders. We developed recombinant endopeptidases and antibodies with a unique brain targeting sequence derived from ApoB and have shown in animal models of AD and PD that these hybrid proteins ameliorate the pathology and recover the functional deficits. These results suggest that the recombinant brain-targeted proteins might be of use in the treatment of AD and PD.






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4:45 Novel Human Resistant Mutant that Acts as Antagonist Reduced Body Weight Gain and Restored Insulin Responsiveness in Mice Fed High Fat DietArieh Gertler, Ph.D., CEO, Protein Laboratories Rehovot; Professor-Emeritus and Head of Research Team, Biochemistry, The Hebrew University of JerusalemResistin promotes both inflammation and insulin resistance associated with energy homeostasis impairment. To block resistin action, we developed a recombinant human resistin mutant (C6A) that acts as a resistin antagonist (RA). We clearly show that RA leads to a significant decrease in body weight of HFD mice. Importantly, RA treatment completely restored glucose tolerance as evidenced by glucose tolerance test and also restored insulin-responsiveness as estimated by insulin tolerance test.

5:15 Local Inhibition of Cytokine Signaling to Treat Anterior and Posterior Ocular DisordersEric Furfine, Ph.D., CSO, Eleven BiotherapeuticsCytokines, chemokines, and growth factors mediate anterior and posterior eye diseases. Our lead product, the IL-1 receptor inhibitor EBI-005, was designed and engineered for the topical treatment of dry eye disease and was biologically active in subjects with dry eye disease. In addition, we engineered an IL-6 inhibitor with potential for local treatment diabetic macular edema. Finally, a novel soluble receptor inhibitor of cytokines IL-17A and IL-17F was engineered for the local treatment of uveitis. Both IL-6- and IL-17-targeted drugs were designed and engineered for intravitreal administration.

5:45-7:00 Welcome Reception in the Exhibit Hall with Poster Viewing

TUESDAY, JANUARY 20

8:00 am Morning Coffee

CONQUERING CANCER

8:30 Chairperson’s RemarksManfred Konrad, Ph.D., Research Director, Enzyme Biochemistry, Max Planck Institute for Biophysical Chemistry

8:35 Late Stage Development of a Chimeric AntibodyManfred Schuster, Ph.D., COO, Apeiron Biologics AGThis case study will focus on how our APN311 chimeric antibody targeting high-risk neuroblastoma was able to overcome clinical, technical and financial hurdles towards our application for market approval planned for mid-2014 in the US and Europe. This talk will also give insights into clinical analytics and our CMC strategy, and highlight a novel immune-therapy administration scheme to reduce side effects and to maintain or even improve clinical response.

9:05 A Recombinant Immunotoxin for Cancer Treatment with Low Immunogenicity by Identification and Silencing of Human T Cell EpitopesRonit Mazor, Ph.D., Research Fellow, Lab for Molecular Biology (LMB), NCI/NIHRecombinant immunotoxins are less active in patients with solid tumors because their immune system makes anti-drug antibodies which inactive the immunotoxin. To suppress the immune response, we have identified and largely silenced the T cell epitopes responsible for the immune response with a redesigned immunotoxin containing T cell epitope mutations that are highly cytotoxic to cells isolated from cancer patients and produces complete remissions in mice with human cancer xenografts.


9:50 Coffee Break in the Exhibit Hall with Poster Viewing

ENGINEERING FLEXIBILITY

11:00 Engineered Affibody Molecules in Multiple Formats for Targeted TherapyFredrik Frejd, Ph.D., Vice President and CSO, Affibody ABThe half-life of biotherapeutics can be extended up to several weeks by applying Affibody’s albumin binding domain (ABD) as a fusion partner. In addition, Antibodies are functionalized using Affibody molecules to create bispecific AffiMabs. Results from oral administration of a half-life extended peptide for treatment of metabolic disease will be shown, along with new data to complement C5-specific and IL-17-specific Affibody-ABD fusion molecules and AffiMabs simultaneously targeting the IL-6 and TNF.

11:30 DeBouganin Fusion Proteins: A “Fit for Purpose” ADC Drug DesignJeannick Cizeau, Ph.D., Associate Director, Research, Viventia Biotech, Inc.Immunotoxins are comprised of a cell targeting domain linked to a cytotoxic toxin payload. DeBouganin, a de-immunized variant of the type I ribosome-inactivating protein (RIP) bouganin, is highly potent and represents an alternative to conventional cytotoxic anti-cancer agents. This presentation will illustrate deBouganin’s distinct mechanisms of action in the context of recombinant fusion proteins and highlight its potency against cancer stem cells and cell lines overexpressing MDR.

12:00 pm Sponsored Presentation (Opportunity Available)

12:30 Session Break

12:45 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own






JANUARY 19-20





























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2:00 BuzZ Session A

3:00 Refreshment Break in the Exhibit Hall with Poster Awards

3:45 BuzZ Session B (Please visit our website for details)

4:45 Close of Conference

4:30-5:00 Short Course Registration

5:00-8:00 Dinner Short Courses See pages 4-5 for details



2nd Annual Enhancing Antibody Binding and Specificity

Emerging Science and New Technologies to Fine-Tune Antibody-Antigen Binding and Target Specificity

JANUARY 21-22PIPELINE 1: PROTEIN ENGINEERING & DEVELOPMENT





























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As the industry expands its repertoire of antibody drug products into new therapeutic areas, product formats and protein constructs, the control of antibody/antigen targeting, binding and specificity will take on a new level of importance for researchers in this field. The second component of the PepTalk Protein Engineering & Development pipeline, the Enhancing Antibody Binding and Specificity conference, presents innovative approaches to the modulation of binding activity, mechanism of action and difficult target challenges such as transmembrane proteins.

TUESDAY, JANUARY 20

1:30 pm Conference Registration



WEDNESDAY, JANUARY 21


SPECIFICITY ENGINEERING

8:15 Chairperson’s Opening RemarksJonas V. Schaefer, Ph.D., Head, High-Throughput Laboratory, Department of Biochemistry, University of Zurich


8:20 The Impact of Anti-IgG Hinge Antibodies in Protease-Enriched DiseasesRobert E. Jordan, Ph.D., Senior Director and Senior Research Fellow (retired), Janssen PharmaceuticalsAnti-IgG hinge antibodies display fine specificity for proteolytically-cleaved IgGs but are not cross-reactive with the intact IgG counterpart. Engagement of anti-hinge antibodies with cell-bound cleaved IgGs restores antibody effector function in vitro and in vivo either when elicited by vaccination or as a mAb. The findings presented here suggest a novel mechanism for enhancing antibody-mediated cell-killing effector functions with application in pathological settings where protease activity is abundant.

9:00 Engineering of High-Affinity Two-in-One Antibodies Using Phage Display Coupled to Deep SequencingSarah Sanowar, Ph.D. Senior Research Associate, Antibody Engineering, Genentech, Inc. – A Member of the Roche GroupTo improve antibody affinity, we sought to develop a robust strategy to identify improved affinity variants beyond traditional phage-based screening. We turned to an approach, which combines affinity-based selection on phage with deep sequencing. Phage libraries with various diversity designs were combined to maximize the searched sequence space. This strategy gave us a comprehensive set of information on the effect of mutations in the antigen-binding site for a two-in-one antibody with dual action Fab for both of its antigens.

STRATEGIES FOR SCREENING LARGE ANTIBODY LIBRARIES

9:30 The Antibiome: Toward Renewable Antibodies to the ProteomeMichael Hornsby, Ph.D. Researcher, Pharmaceutical Chemistry, University of California, San FranciscoWe have developed a robust high-throughput robotic pipeline for the generation and validation of renewable recombinant antibodies utilizing antibody phage-display. In order to match the capacity of the antibody generation to the availability of input antigens, we have developed several robust antigen production pipelines. Both pipelines working together in concert will allow the Recombinant Antibody Network (RAN) to develop quality renewable antibody reagents to the proteome.


10:50 Pipeline 2.0: Integrating All Aspects from Target Acquisition through Binder Validation for Optimized Binder GenerationJonas V. Schaefer, Ph.D., Head, High-Throughput Laboratory, Department of Biochemistry, University of ZurichA robust pipeline for the high-throughput generation of affinity reagents enables many scientific projects and novel applications. To optimize the pipeline’s throughput, our laboratory developed a streamlined process, consisting of parallel Ribosome Display selections and various semi-automated high-throughput screenings. Also including aspects of target acquisition to binder validation while decreasing its time and cost requirements, we perform simultaneous selections against 94 targets and screen and validate several thousand binders in parallel for their characteristics.

2:00 BuzZ Session A




































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SCREENING FOR RARE ANTIBODIES

11:20 An in vivo Human-Plasmablast Enrichment Technique Allows Rapid Identification of Therapeutic Influenza A AntibodiesGerald Nakamura, Ph.D., Scientific Manager, Antibody Engineering, Genentech, Inc. – A Member of the Roche GroupRecent advances enabling the cloning of human immunoglobulin-G genes have proven effective for discovering monoclonal antibodies with therapeutic potential. However, these antibody-discovery methods are often arduous and identify only a few candidates from numerous antibody-secreting cells. We describe an in vivo enrichment technique that identified broadly influenza neutralizing human antibodies with high frequency. Using this technology, we identified four broadly neutralizing influenza A antibodies by screening only 840 human antibodies.

11:50 Towards a Quantum Leap in the Utility of Combinatorial Libraries for Drug Hunters by Placing New Functional Screening Options Up FrontRonald M. Lindsay, Ph.D., CEO, Zebra BiologicsWhereas the relative merits of deriving therapeutic antibody candidates via humanized mouse or phage display approaches are still debated, both approaches yield an over abundance of initial ‘hits’ as defined by binding to a desired target. Selecting “winners” from these initial hit binders remains a bottleneck. I will discuss new screening approaches that allow more direct high throughput selection of functional antibodies for known targets and the discovery of new targets.

12:20 pm Recent Advances and Successes in Structure-Based Protein Design Using BioLuminate

Sponsored by

David A. Pearlman, Ph.D., Senior Principal Scientist, SchrödingerWe describe several recent studies demonstrating the power of theoretical structure-based protein design tools as applied to antibodies, enzymes, and other proteins. We demonstrate how these tools can be applied to critical issues in design such as improved affinity and stability, as well as the elucidation of hot spots in an attempt to triage research efforts toward residues where modifications are more likely to be successful.

12:50 Session Break


RATIONAL APPROACHES TO ANTIBODY ENGINEERING

2:00 Chairperson’s RemarksMichael Drummond, Ph.D., Applications Scientist, Chemical Computing Group

2:05 ADAPT – A Platform for Antibody Affinity MaturationTraian Sulea, Ph.D., Senior Research Officer, Human Health Therapeutics, National Research Council CanadaTo assist affinity maturation of therapeutic antibodies we have implemented a platform combining binding affinity predictions with stepwise experimental verification. Starting from the crystal structure of an antibody-antigen complex, an efficient workflow based on mutation additivity interleaves ADAPT predictions with experimental validation from single-point mutations up to typically quadruple mutants. In this talk we will present prospective examples of employing ADAPT to affinity mature antibodies against VEGF and HER2.

2:35 Extracting Life’s Operating Manual – Comprehensive Ruleset Discovery and Bioengineering ApplicationsJacob Glanville, Ph.D., CSO, Distributed BioWithout comprehensive rulesets to guide rational design, most antibody bioengineering efforts are iterative process of applying and testing small changes. The combination of high-throughput sequencing, combinatorial gene synthesis and selection pressures provides a unique opportunity to enumerate the ruleset space that governs a molecule or entire repertoire. We’ll review the last 6 years of theory and practical application, then transition to highlight some of the startling novel technological applications such rule-based design can enable.

3:05 Precise and Efficient Antibody Epitope Determination Through Library Design, Yeast Surface Display and Next- Generation SequencingThomas Van Blarcom, Ph.D., Principal Scientist, Protein Engineering, Pfizer, Inc.Here we describe a method to precisely and efficiently map the epitopes of small panels of antibodies in parallel over the course of several weeks. This method relies on the nexus of rational library design, quantitative yeast surface display and next generation DNA sequencing and was demonstrated by mapping the epitopes of several antibodies that neutralize the alpha toxin from Staphylococcus aureus.

3:35 Creating Focused Mutant Libraries for Protein Engineering Sponsored by

Michael Drummond, Ph.D., Applications Scientist, Chemical Computing GroupProtein engineering plays a pivotal role in modulating the function, activity and physical properties of biologics. Representative strategies employed in protein engineering include directed evolution and rational protein design. Although both approaches are effective at identifying and optimizing protein therapeutic candidates, efficient search and evaluation of an excessively large sequence design space becomes challenging and requires multiple experimental rounds to reasonably assess the sequence space. Here we have developed a computational approach which predicts mutation probabilities for given residue sites in specified sequences. In assessing the probabilities at given residue sites, the sequence search space can be efficiently sampled to design and produce focused mutant libraries.



































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»4:30 PLENARY KEYNOTE SESSION See page 2 for details

5:30-7:00 Reception in the Exhibit Hall with Poster Viewing

THURSDAY, JANUARY 22


DISCOVERY AND DEVELOPMENT OF ANTIBODIES FOR MEMBRANE PROTEIN TARGETS

8:30 Chairperson’s RemarksTrevor Wilkinson, Ph.D., Associate Director, Protein Sciences, Antibody Discovery and Protein Engineering, MedImmune

8:35 GPCR Expression by Individual Cell Types: Novel Membrane Targets for Therapeutic AntibodiesPaul Insel, Ph.D., Professor, Pharmacology & Medicine, University of California, San DiegoGPCRs, the largest family of membrane receptors, also represent the largest class of targets of FDA-approved drugs. However, little is known regarding GPCR expression by individual cell types. Using a GPCRomic strategy, we have identified the full range of non-chemosensory GPCRs, including numerous orphan GPCRs, expressed by many such cell types. Our other findings suggest the possibility of using antibody therapeutics directed at such receptors.

9:05 Targeting T Cells with an Anti-Ion Channel Antibody with Ultralong CDR H3sVaughn Smider, M.D., Ph.D., Assistant Professor, Molecular Biology, The Scripps Research InstituteThe relatively flat binding surface of a typical antibody paratope may not allow optimal interactions with certain epitopes. Cow antibodies contain ultralong CDR H3’s consisting of a b-ribbon stalk and disulfide-bonded knob. The knob structures are reminiscent of ion channel bioactive peptides known to interact with high specificity and affinity with ion channels. We have engineered a cow antibody to bind and inhibit an ion channel critical for T cell activation in autoimmune disease and inflammation.


9:50 Coffee Break in the Exhibit Hall with Poster Awards

10:50 Strategies to Obtain Antibodies to Difficult Membrane Protein TargetsJohn S. Kenney, Ph.D. President & CEO, Antibody Solutions Some membrane protein targets, including high homology proteins, G-protein-coupled-receptors (GPCRs), Ion Channels, and Multicomponent Receptor complexes present unique challenges in the generation of specific, high-affinity, and functional antibodies. Each target requires a custom approach based upon the nature of the target and the desired characteristics of the antibody. Strategies for antigen design, immunization, and screening of antibodies to difficult membrane proteins and results obtained will be presented.

11:20 Discovery and Optimization of Novel Anti G-Protein Coupled Receptor Monoclonal AntibodiesTrevor Wilkinson, Ph.D., Associate Director, Protein Sciences, Antibody Discovery and Protein Engineering, MedImmuneG-protein coupled receptors represent a challenging target class for the isolation and optimization of therapeutic biologics. We have used a combination of immunization and phage display to isolate functional antagonistic antibodies targeting a chemokine receptor and a formyl peptide receptor that will be presented as case studies. We also describe how combinatorial mutagenesis approaches have been used to make significant improvements to both affinity and species cross-reactivity of a lead molecule and demonstrate that the optimised antibodies show significantly increased potency in cellular disease assays.

11:50 High-Throughput Strategies to Obtain High Affinity, Specific, and Conformationally Selective Recombinant Antibodies to Membrane Proteins by Phage DisplayMarcin Paduch, Ph.D., Technical Director, Synthetic Antibody & Crystallography Core Facility, The University of ChicagoState of the art methods for generating recombinant antibodies to membrane proteins require the use of detergents that do not necessarily mimic the native lipid environment. We have developed a suite of next-generation high-throughput technologies to generate high affinity, specific, and conformationally selective reagents by exploiting liposomes, nanodiscs and cell surface display for antigen presentation. These native-like environments create the possibility of trapping physiological states otherwise not accessible by current methods.

12:20 pm Session Break





2nd Annual Improving the Clinical Efficacy of Antibody Therapeutics

Cutting-Edge Protein Engineering for the Next Generation of Safe and Effective Biotherapeutics






























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When the more than 350 therapeutic monoclonal antibodies in development advance into the clinic and to commercial launch, the quality of therapeutic response will become increasingly important to regulatory agencies and frugal payors. Regulators are demanding better data to support claims of safety and potency – and payors are seeking meaningful therapeutic benefits relative to existing standards of care before adding higher-cost biotherapeutics to formularies. The Improving the Clinical Efficacy of Antibody Therapeutics conference showcases state-of-the-art discovery and development-stage engineering strategies for improving the safety and effectiveness of this important class of biologic drugs.


11:30 am Conference Registration

12:30 pm Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own

1:30 Ice Cream Break in the Exhibit Hall with Poster Viewing

GLYCOENGINEERING AND ENHANCING EFFECTOR FUNCTION

2:00 Chairperson’s Opening RemarksJanine Schuurman, Ph.D., Vice President, Research, Genmab


2:05 A Palette of Engineered Fc Domains for Optimal Antibody-Mediated Target Cell Clearing and ImmunomodulationGeorge Georgiou, Ph.D., Professor, Cockrell Family Regent’s Chair in Engineering, Department of Biomedical Engineering, The University of Texas at AustinWe have engineered a variety of aglycosylated Fc domains displaying: (i) very high binding affinity and selectivity for each individual human Fcγ receptor; (ii) Fc domains that bind only to C1q; (iii) to Fcγ receptors as well the FcγRI receptor that binds to IgA. These Fc domains were shown to elicit unique profiles of immune cell activation and the clearance of target cells.

2:45 Improving the Therapeutic Efficacy of pH and Calcium-Dependent Antigen Binding AntibodiesTomoyuki Igawa, Ph.D., Group Manager, Discovery Research, Chugai Pharmaceutical CompanypH or calcium-dependent antigen binding antibody enhances antigen elimination by dissociating the antigen in the endosome. Here we report that Fc receptors such as FcRn and inhibitory FcgRIIB can be exploited to further accelerate the antigen elimination by pH or calcium-dependent antigen binding antibody. Enhancing binding to Fc receptors, either by Fc engineering or by formation of multimeric antibody-antigen complex, significantly accelerated antigen elimination from plasma in vivo.

3:15 Addressing Challenges to Transporting Antibodies Across the Blood-Brain BarrierJames A. Ernst Ph.D. Senior Scientist, gRED, Genentech

3:45 Selected Poster Presentation: Optimization of Antibody Fc Effector Functions by In Vitro GlycoengineeringRoland Dorn, International Product Manager, Roche Diagnostics GmbH

4:15 Refreshment Break in the Exhibit Hall with Poster Viewing

5:00 Complement Is Activated by IgG Hexamers Assembled at the Cell SurfaceJanine Schuurman, Ph.D., Vice President, Research, GenmabComplement activation by antibodies is an important mechanism in immune defense and immunotherapy. Using X-ray crystallography, mutagenesis studies and cryo-EM tomography, we revealed that IgG antibodies form hexamers on the cell surface following antigen binding. Enhancing hexamerisation on the cell surface by using the HexaBodyTM platform potentiated the intrinsic killing capability of antibodies in in vitro, in vivo and ex vivo models.

5:30 Cytotoxic Mechanisms of Immunotherapy: Harnessing Complement in the Action of Anti-Tumor Monoclonal AntibodiesRonald P. Taylor, Ph.D., Professor of Biochemistry, University of VirginiaWe followed CDC mediated by CD20 mAbs engineered to enhance Fc-Fc contacts, thus promoting strong C1q binding and rapid CDC. Confocal microscopy movies revealed that during CDC, Ca2+ rapidly enters cells and is soon localized to mitochondria. Ca2+ poisoning likely is the most proximate mediator of cytotoxicity. These observations should allow for deeper understanding of CDC mechanisms, and will play a critical role in development of more effective immunotherapeutic mAbs.

6:00-7:00 Reception at the Tiki Pavilion

FRIDAY, JANUARY 23


COMBINATION STRATEGIES FOR ENHANCING THE EFFICACY OF ANTIBODY THERAPY

9:00 Chairperson’s RemarksYasmina Abdiche, Ph.D., Research Fellow, Rinat-Pfizer

9:05 Using an Oligoclonal Approach to Target HER3/ErbB3Matthew Robinson, Ph.D., Assistant Professor, Fox Chase Cancer CenterHER3/ERBB3 is recognized as an important therapeutic target in a variety of cancers and clinical validation of antibody-based therapies targeting this receptor is currently underway. We have taken an oligoclonal approach to develop an optimized anti-HER3/ERBB3 agent capable of inhibiting signaling through this critical receptor and its heterodimeric partners. Work detailing the isolation and characterization of an anti-HER3/ERBB3 oligoclonal will be discussed.


































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9:35 Development of an Antibody Combination Therapeutic (ACT) for the Treatment of Ventilator Associated PneumoniaElizabeth Reczek, Ph.D., President and CSO, Excelimmune, Inc.Antibody Combination Therapeutics (ACTs) are a novel class of polyvalent biopharmaceuticals, uniquely suited for the treatment of complex diseases. Excelimmune has developed a fully human antibody discovery platform and AAV-based, virus-free recombinant protein expression technology capable of rapid and consistent production of complex antibody mixtures in a single batch format. We are leveraging this technology to develop an ACT therapeutic for the treatment of Ventilator Associated Pneumonia (VAP).


CHARACTERIZATION OF PROPERTIES IMPACTING EFFICACY

11:00 Analytical Methods to Characterize the Binding Kinetics, Affinity, and Epitope of Therapeutic AntibodiesYasmina Abdiche, Ph.D., Research Fellow, Rinat-PfizerThis talk will focus on leveraging the strengths of the commercially available repertoire of label-free biosensor technologies in the context of discovering therapeutic monoclonal antibodies. We will show examples of several types of assays that can be used to characterize large panels of antibodies and guide the selection of those with appropriate on-target affinity and specificity.

11:30 Impact of Effector Cells on in vitro ADCC Activity of Therapeutic AntibodiesShan Chung, Ph.D., Senior Scientist, Bioanalytical Sciences, Genentech, Inc. – A Member of the Roche GroupIn this study we evaluated the impact of different type of effector cells on the in vitro antibody-dependent cell-mediated cytotoxicity (ADCC) activity of two glycoforms of a humanized IgG1 antibody. The results of this study show differential effects on both the efficacy and potency of the antibodies by different effector cells and that both the allotype and the expression level of CD16a affect the potency of effector cells in ADCC assays.

12:00 pm Analytical Fc Receptor Affinity Chromatography for Functional Characterization of Monoclonal AntibodiesTilman Schlothauer, Ph.D., Senior Scientist, Protein Analytics, Roche Diagnostics GmbHFc receptor-based affinity chromatography is a new emerging field of Fc functionality analytics. Until now different human FcγReceptors (FcγRIIIa & IIa) and the Fc Receptor neonatal (FcRn) from three species have been utilized for coupling onto Sepharose-based matrices. FcRn affinity columns separate antibody species (analytical and preparative) that differ in their affinity to FcRn receptors, using conditions that closely resemble the physiological mechanism of interaction between IgG and FcRn.


1:00 Session Break


STRATEGIES FOR IMPROVING THE EFFICACY OF ANTIBODY THERAPEUTICS

2:00 Chairperson’s RemarksJon Wojciak, Ph.D., Scientist, Lpath

2:05 Monoclonal Antibody Therapy for Multiple MyelomaFrits van Rhee, M.D., Ph.D., Professor, Medicine and Director, Developmental and Translational Medicine, Myeloma Institute for Research and Therapy, University of Arkansas

2:35 Observations Regarding Antibody Pharmacokinetics and a Case Study; Engineering a mAb for Prolonged Half-Life to Better Assess an Animal Model SpeciesTom Nesspor, Research Scientist, Biologics, JanssenEpitope, affinity, immune effector function, and pharmacokinetics determine the efficacy of therapeutic mAbs. This talk will review recent findings in our group relating to antibody pharmacokinetics such as non-FcRn influences on clearance, strategies for prolongation and reduction of half-life, and correlations between human FcRn transgenic mice and human pharmacokinetics. It will conclude with a case study describing the engineering of mAbs for prolonged half-life to better assess an important animal model species.

3:05 Antibodies that Target Bioactive LipidsJon Wojciak, Ph.D., Scientist, LpathDeveloping therapeutic antibodies with favorable biophysical properties (e.g. antigen affinity and specificity, solubility, aggregation, etc.) is a formidable challenge, and engineering these antibodies can lead to molecules that undergo rapid-reversible, self-association. Using molecular modeling and site-directed mutagenesis, the reversible self-association of a humanized, monoclonal anti-lipid antibody was minimized while the antigen affinity and specificity was retained.






SUBMIT A POSTER

Cambridge Healthtech Institute encourages attendees to gain further exposure by presenting their work in the poster sessions.

Reasons you should present your research poster at this conference:

• Your poster will be exposed to our international delegation• Receive $50 off your registration• Your poster abstract will be published in our conference

materials• You will automatically be entered into the poster competition• Your research will be seen by leaders from top

pharmaceutical, biotech, academic and government institutes

To secure a poster board and inclusion in the conference materials, your abstract must be submitted, approved and your registration paid in full by November 21, 2014.





























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JANUARY 22-23

3:35 Computational and Experimental Mapping of Deimmunized Biotherapeutic Design SpaceKarl E. Griswold, Ph.D., Associate Professor, Bioengineering, Thayer School of Engineering, Dartmouth CollegeBiotherapeutics are powerful drugs, but the risk of protein immunogenicity represents a barrier to their broader development and use. While methods for T cell epitope identification are maturing rapidly, facile selection of deimmunizing yet function-preserving mutations remains a challenge for protein engineers. Here we describe experimental validation of integrated deimmunization algorithms that simultaneously optimize both protein function and immunogenic potential. We demonstrate the capacity to tune a protein’s sequence-structure-function-immunogenicity relationships.




PIPELINE 2: ANTIBODY THERAPEUTICS

Inaugural Cancer Targets for Antibody Therapeutics

Discovery, Engineering and Optimization of Next-Generation Oncology Targets

JANUARY 19-20





























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The science and technology of antibody engineering have brought forth a new era of therapeutic antibodies in oncology, with new product formats and an intense interest in immune system modulation now at the forefront of many company development efforts. PepTalk’s new Cancer Targets for Antibody Therapeutics conference explores new directions in the discovery of emerging and challenging targets in this space – and the steps that will be required in developing these into next-generation therapeutics for patients.

SUNDAY, JANUARY 18




MONDAY, JANUARY 19


NEW APPROACHES TO TARGETING THE TUMOR MICROENVIRONMENT

9:00 Chairperson’s Opening RemarksGregory P. Adams, Ph.D., Co-Leader, Developmental Therapeutics Program, Fox Chase Cancer Center


9:10 Multifunctional Angiogenesis Inhibitors Designed from Non-Antibody ScaffoldsJennifer R. Cochran, Ph.D., Associate Professor, Bioengineering and Chemical Engineering, Stanford UniversityWe engineer protein therapeutics, based on growth factor ligands and receptors with altered biochemical and biophysical properties, as alternatives to antibodies. I will present recent work where we used a growth factor as a scaffold to create ligand-based antagonists that bind to multiple cell surface receptors, and demonstrated that these proteins more effectively inhibit angiogenic processes compared to mono-specific receptor targeting agents.

9:50 Manipulating the Tumor Microenvironment to Enhance Effector Function for Improved Antibody Efficacy in PatientsStephen Beers, Ph.D., Associate Professor, Antibody & Vaccine Group, University of SouthamptonSuccessful antibody therapy appears to rely predominantly on Fcγ receptor expressing effector cells such as macrophages. Unfortunately, a number of cancers have proven resistant to antibody therapy potentially due to the adverse effects of the tumor microenvironment on these cells. Here, the potential of harnessing tumor associated macrophages as effectors will be discussed as well as means presented by which they may be re-programmed to enhance their antibody effector capacity.

10:20 Coffee Break

ENGINEERING ANTIBODIES FOR IMPROVED TUMOR PENETRATION

10:45 A Cell-Penetrating Antibody Technology Platform: Making the Undruggable DruggableHua Eleanor Yu, Ph.D., Billy and Audrey L. Wilder Endowed Professor of Tumor Immunology, Co-Leader of Cancer Immunotherapeutics Program, City of Hope Comprehensive Cancer CenterWe have developed a novel technology platform to allow efficient cell-penetration of proteins/antibodies in vitro and in vivo. Using flow cytometry, confocal imaging and Western blotting, we demonstrate the self-penetrating ability of the modified antibodies. Both local and systemic administrations of the modified antibodies effectively inhibit their intracellular targets in tumors, resulting in tumor cell apoptosis and tumor regression in multiple models. Our discoveries enable the development of a new class of research tools and novel therapeutics.

11:15 The Effect of Molecular Weight, PK, and Valency on Tumor Biodistribution and Efficacy of Antibody-Based DrugsRuth Muchekehu, Ph.D. Research Scientist, Vertex PharmaceuticalsTo explore the role of pharmacokinetics, valency, and size on tumor targeting, the biodistribution of an FGFR4 targeting CovX-body (an FGFR4-binding peptide linked to a non-targeting IgG scaffold; 150 kDa), F(ab)2 (100 kDa) and Fab (50 kDa) fragments was measured. The highest percent of injected drug was achieved with the IgG, and increasing the valency of the IgG by conjugating a homodimeric peptide to the scaffold, translated into superior efficacy.

11:45 How to Leverage Oncogene Addiction: Targeted Biological Therapy Inducing Growth Factor Receptor Internalization and DegradationJohn Haurum, M.D., D.Phil., CEO, F-star GmbH & F-star Biotechnology Ltd.FS102 is a HER2-specific Fcab™ (Fc with antigen binding). In HER2-overexpressing tumour cells, FS102 induce profound HER2 degradation and apoptosis, and FS102 eliminates HER2-overexpressing tumours in patient-derived mouse xenograft models. We present this as an example of a general class of oncogene-targeted biological therapies, which induce tumour killing via internalization and degradation of the addictive growth factor receptor.


12:45 Session Break







JANUARY 19-20





























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EMERGING TARGETS

2:00 Chairperson’s RemarksJohn Haurum, M.D., D.Phil., CEO, F-star GmbH & F-star Biotechnology Ltd.

2:05 Targets for Antibodies in Neuro-Oncology: Getting Past the Blood-Brain BarrierLois Lampson, Ph.D., Associate Professor of Surgery, Brigham and Women’s HospitalAntibody-mediated therapy for brain tumors is thought to face two hurdles: The blood-brain barrier (BBB) and, for some effector mechanisms, the presumed “immune privilege” of the brain. Here we ask, for different kinds of brain tumor targets: Are the BBB or “privilege” really the key problems? How different is the brain, really, from other sites?

2:35 New Anti-Cancer Immunotherapy Strategies Utilizing Validated Tumor TargetsBarbara Swanson, Ph.D., Director, Research, Sorrento Therapeutics, Inc.

3:05 SAR650984, A CD38 Monoclonal Antibody for Selected CD38+ Hematological MalignanciesFrancisco Adrian, Ph.D., Section Head, Sanofi OncologyCD38 is a type II transmembrane glycoprotein highly expressed at the surface of malignant multiple myeloma plasma cells. SAR650984 is a humanized IgG1 antibody targeting CD38 in PhII clinical trials. The preclinical characterization of SAR650984 will be presented, including epitope mapping, impact on CD38 enzymatic activity, pro-apoptotic activity in MM cellular models and patient samples, and in vivo activity in combination with bortezomib.

3:35 Fluorescent Human Synthetic Nano-antibodies (snAbs) Highlight Specifically Cancer Cells to Guide Oncology Surgeons in Efficient Removal of Cancerous TumorsMarek Malecki, M.D., Ph.D., CEO, Phoenix Biomolecular Engineering FoundationThe most difficult problem, for surgeons pursuing removal of cancerous tumors, is to determine the borders of the tumors, so that they can remove cancerous tissue, while minimizing iatrogenic injury to the healthy tissue. To resolve this problem, we designed and manufactured fluorescent, human, synthetic nano-antibodies (snAbs) against cancer cells. The cancer cells were specifically, strongly, and permanently highlighted with our fluorescent snAbs, while making them easy to distinguish from healthy cells in vivo.


4:15 Development of a Tumor Specific, CTLA4 AntagonistJohn C. Williams, Ph.D., Associate Professor, Molecular Medicine, City of Hope

4:45 Drugging the Undruggable: Using Knowledge-Based Design to Develop Antibodies against Difficult TargetsGregory P. Adams, Ph.D., Co-Leader, Developmental Therapeutics Program, Fox Chase Cancer CenterThe development of new clinically relevant antibodies has historically depended upon the immunization of animals or the selection of clones with desired specificity from large antibody libraries. While this works for many targets there are numerous important target epitopes that are difficult or even “undruggable”. Working with collaborators we have pursued a rational design approach employing structure prediction, loop grafting and computational combinatorial CDR display to develop antibodies specific for difficult targets.

5:15 Selection of Antibodies for T Cell Redirected KillingDiego Ellerman, Senior Research Associate, Protein Chemistry, Genentech, Inc. – A Member of the Roche Group T cell recruitment and redirected killing is a growing clinical strategy that is currently being explored for different oncological targets. This approach requires the use of bispecific antibodies targeting both the T cell receptor and a tumor-specific antigen. Different antibodies’ properties could play a role in determining the efficacy of the molecule, such as affinity, epitope location, binding geometry. We present case studies showing the influence of these factors.


TUESDAY, JANUARY 20


DISCOVERY AND ENGINEERING OF IMMUNOMODULATORY ANTIBODIES

8:30 Chairperson’s RemarksDavid King, Ph.D., CSO, AnaptysBio, Inc.

»FEATURED PRESENTATION8:35 Monoclonal Antibodies as the Foundation of the Immunotherapy RevolutionDavid Meininger, Ph.D., MBA, Executive Director, Business Development & Licensing, Merck & Co., Inc.Antibody-mediated inhibition of the PD-1 checkpoint pathway embodies a paradigm shift in cancer therapy. However, nearly half of melanoma patients and a majority in other indications with clinical data published to date have failed to respond to therapy. The highest priorities in cancer research today are arguably understanding PD-1 therapy non-responsiveness and developing new PD-1 alternatives and combinations with antibodies anticipated to represent foundational components of associated emerging treatment regimens.






JANUARY 19-20





























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9:05 Engineering a Novel, Multivalent Fusion Protein that Potently Agonizes a TNFSF ReceptorMelissa Damschroder, Ph.D., Associate Director, Research & Development, Antibody Discovery and Protein Engineering, MedImmuneAlthough soluble recombinant OX40L assembles into trimers and binds to OX40, it only produces a weak co-stimulatory signal. In order to generate a recombinant therapeutic with enhanced immune modulating properties, we have engineered a multivalent OX40 ligand fusion protein to agonize the trimeric OX40 receptor. The soluble hexameric OX40L “dimer of trimers” binds to OX40 on T cells and potently mimics the activity of membrane bound OX40 ligand/receptor signaling. Extensive functional and physicochemical characterization of the hexameric protein was performed to ensure its developability and manufacturability.

9:35 Selected Poster Presentation: High Throughput SPR for First-Pass Kinetics™ Screening of Antibody TherapeuticsJosh Eckman, CEO & President, Wasatch Microfluidics


11:00 Control of Regulatory T (Treg) Cell Function by Protein Kinase C-eta (PKCη): A Novel Target for Cancer ImmunotherapyAmnon Altman, Ph.D. Director, Scientific Affairs, Division of Cellular Biology, La Jolla Institute for Allergy and ImmunologyAntibody-mediated blockade of the checkpoint inhibitory receptor, CTLA-4, is currently used for treating patients suffering from melanoma and other cancers. Treg cells represent an important immune escape mechanism that facilitates tumor growth. I will report on a novel Treg cell-intrinsic signaling pathway mediated by recruitment of PKCη to the CTLA-4 cytoplasmic tail (Nat. Immunol. 15:465, 2014). In the absence of this pathway, Treg cells are unable to suppress anti-tumor immunity.

11:30 Discovery and Engineering of Novel Antibodies to Immune CheckpointsDavid J. King, Ph.D., CSO, AnaptysBio, Inc.Among the most promising approaches to cancer therapy is the activation of anti-tumor immunity by blockade of immune checkpoints such as CTLA-4 and PD-1. A number of other immune checkpoints are of interest, and functional antibodies to PD-1, TIM-3 and LAG-3 have been generated. Inhibition of each pathway demonstrates activity, and combinations can increase T cell activation and have the potential to lead to increased clinical efficacy.


12:30 Session Break


2:00 BuzZ Session A









2nd Annual Antibody-Drug Conjugates

Engineering Targeted Therapeutics

JANUARY 21-22





























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The therapeutic potential of antibodies is enhanced by conjugating them to small molecule drugs. This combination merges the benefits of highly potent drugs with selective binders of specific tumor antigens. Antibody-drug conjugates offer the promise of delivering more powerful tumor-killing activity while resulting in diminished side effects for cancer patients. This important Antibody-Drug Conjugates conference brings together leaders in the world of ADCs who share their R&D case studies, along with their preclinical and clinical data, to illustrate how this form of empowered antibody is transforming next-generation antibody therapeutics.

TUESDAY, JANUARY 20


2:00 BuzZ Session A







MOVING ADCs SAFELY INTO THE CLINIC

8:15 Chairperson’s Opening RemarksTrevor Hallam, Ph.D., CSO, Sutro Biopharma, Inc.


8:20 Examination of ADC Safety Challenges in the Clinical SettingFlavia Brunstein, M.D., Ph.D., Safety Science Leader, Safety Risk Management, Genentech, Inc. – A Member of the Roche GroupThe concept of an ADC is to improve the therapeutic window of cancer chemotherapy, through targeted delivery of highly potent cytotoxic molecules directly to tumor cells expressing unique antigens that are specific to the monoclonal antibody. Preliminary clinical data are encouraging, but toxicity still occurs.

9:00 In vitro-in vivo Molecular Integrity of Antibody-Drug Conjugates: Applying Learning to Clinical Measurements for ADCsDan Rock, Ph.D., Scientific Director, Pharmacokinetics and Drug Metabolism, Amgen, Inc.Characterizing the mechanisms of ADC instability and release of free cytotoxin are germane in the design of the next generation of ADCs. The methods and analytical tools useful in characterizing the ADME and stability of ADCs will be reviewed.

9:30 Engineering Antibodies and ADCs for Successful DevelopmentLars Linden, Ph.D., Group Leader and Head, Protein Biochemistry, Bayer HealthCareDevelopability analysis of antibodies and ADCs determines, together with cell line productivity and cost-of-goods analysis, the manufacturing feasibility of a drug candidate. A thorough biochemical & biophysical characterization is performed to analyze the intrinsic stability and technical robustness of clinical candidates. Standardization is ensured by a check of antibody platform compatibility (DSP and analytics). Early buffer screening is performed for accelerated formulation development.


MODELING AND ENGINEERING BREAKTHROUGHS

10:50 Mechanistic Models for Designing Efficacious ADCsArijit Chakravarty, Ph.D., Director, Modeling & Simulation, Takeda Pharmaceuticals International Co.

11:20 Bioconjugation Strategies for Generating Serum-Stable Antibody Conjugates James T. Patterson, Ph.D., Scientist, Ferring Research InstituteBioconjugation strategies commonly rely on maleimide linkers to react with cysteine thiols for the synthesis of antibody-drug conjugates. However, thioether exchange with serum proteins and other metabolites can compromise conjugate stability and in vivo efficacy. We have developed a phenyloxadiazole sulfone linker for the preparation of antibody conjugates. This sulfone linker site-specifically labeled engineered cysteine residues in THIOMABs and SELENOMABs as well as improved antibody conjugate stability in human plasma at sites previously shown to be labile for maleimide conjugates.






JANUARY 21-22





























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11:50 Novel Bifunctional Linkers for the Synthesis of Homogeneous ADCsDavid Jackson, Ph.D., Principal Scientist, Igenica, Inc.Igenica has developed novel bifunctional linkers that enable the synthesis of homogeneous ADCs with an optimal DAR of 4 drugs per antibody. The linkers are compatible with a variety of payloads and no antibody engineering is required. Examples of homogeneous ADCs made with different antibodies and novel payloads will be presented and compared to their heterogeneous ADC counterparts made with conventional linkers. Supporting data will also be discussed.


12:50 Session Break


ADCs FOR ONCOLOGY

2:00 Chairperson’s RemarksAdeela Kamal, Ph.D., Associate Director, Oncology Research, MedImmune

2:05 An EGFR Targeting Antibody-Drug Conjugate Engineered for Increased Tumor SpecificityChristopher D. Thanos, Ph.D., Director, New Molecular Entities, Halozyme Therapeutics, Inc.Cancers with activating KRAS/BRAF mutations and EGFR+ tumor genotypes are resistant to inactivation by EGFR targeting agents and correspond to a significant unmet medical need. We hypothesized that the cytotoxic mechanism of action of anti-EGFR ADC could be active against these tumor types. We engineered an anti-EGFR monoclonal antibody with significant binding to EGFR+ tumors and attenuated binding to human skin in a mouse xenograft to target these intractable tumor types.

2:35 Potent and Selective Alpha-Amanitin-Antibody ConjugatesBrian Mendelsohn, Ph.D., Senior Scientist, ADC Chemistry Group Leader, Agensys, Inc.We have developed a technology based on arming antibodies with derivatives of the rigid, water-soluble bicycle-octapeptide alpha-amanitin, a highly potent inhibitor of eukaryotic RNA polymerase II. We have shown in various in vitro and in vivo models, with antibodies to several different oncology-related antigens, the tumor-selective activity and high potency of these ADCs. We have explored and elucidated the in vivo metabolite-profiles, as well as performed toxicological studies.

3:05 Brain-Penetrant Peptide-ADCs Reduce Tumor Size and Increase Survival in Mice with HER2-Positive Brain TumorsJean Lachowicz, Ph.D., CSO, Angiochem, Inc.Using a peptide recognized by the brain capillary endothelial cell receptor LRP1, peptide-mAb conjugates can access the brain. Applying this strategy to ADCs has produced the first examples of ADCs entering the brain by receptor-mediated transcytosis. Targeting the brain tumor cells, these therapeutic agents release their toxic payload, resulting in a significant decrease in tumor size and increase in survival.

3:35 INTERACTIVE PANEL DISCUSSIONModerator: Trevor Hallam, Ph.D., CSO, Sutro Biopharma, Inc.Panelists Include:Jean Lachowicz, Ph.D., CSO, Angiochem, Inc. John Lambert, Ph.D., Executive Vice President and CSO, ImmunoGen, Inc. David Jackson, Ph.D., Principal Scientist, Igenica, Inc. o Overcoming Current Challenges o Trends & Technologies o The Future of ADC Development






CONJUGATION AND PAYLOAD TECHNOLOGY

8:30 Chairperson’s RemarksChristopher D. Thanos, Ph.D., Director, New Molecular Entities, Halozyme Therapeutics, Inc.

8:35 Building Effective ADCs: Selecting the Right Target, Antibody and PayloadAdeela Kamal, Ph.D., Associate Director, Oncology Research, MedImmune, LLCAntibody drug conjugates (ADCs) combine the specificity of antibodies with the potency of small molecule cytotoxic drugs. The four key components of building an effective ADC is the tumor target, the antibody, the linker and the cytotoxic drug. This talk will review our strategies for identifying high value tumor targets, applying state-of-the art ADC technology with novel payloads and site-specific conjugation, and advancing ADC programs to the clinic.






JANUARY 21-22





























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9:05 Innovative Payload Platform Powers ADC DevelopmentJohn Lambert, Ph.D., Executive Vice President and CSO, ImmunoGen, Inc.ADC therapeutics utilizing potent tubulin-acting agents are an important new class of antibody-based anticancer agents. There are two such agents, brentuximab vedotin and ado-trastuzumab emtansine, approved for treatment of cancer patients, and more than 30 others in clinical testing. Building on these successful ADC platforms are ADCs containing payloads with different killing mechanisms. Exciting advances in development of potent DNA-interacting agents will be discussed.

9:35 Q & A with Speakers


10:50 Delivery of Potent Cytotoxins as ADC PayloadsVishal Verma, Ph.D., Scientist, Medicinal Chemistry, Genentech, Inc. – A Member of the Roche GroupThe vast majority of ADCs approved for use and in the clinic utilize derivatives of maytansine and auristatin. While the use of these cytotoxic agents has proven successful, the next generation of ADCs has the opportunity to make use of additional payloads with the potential to address current shortcomings. The identification and evaluation of novel ADC payloads will be discussed.

11:20 Best-in-Class ADCs; Homogeneity is Necessary but Not Sufficient - Selection of Optimal Conjugation Site is ParamountTrevor Hallam, Ph.D., CSO, Sutro Biopharma, Inc.Homogeneity of ADCs is necessary, but insufficient alone, to assure best-in-class; conjugation positional analysis is key. Cell-free protein synthesis offers fundamental advantages for design and efficient manufacture, while new classes of conjugated warhead combinations show great promise. Systematic analysis of linker warhead positioning reveals key differences in solid tumor killing potency in vivo that are not predicted by internalization rates, binding affinity, stability or PK, and may reflect differences in tumor drug disposition.

11:50 Conventional and Site-Specific Approaches to Enable New Classes of ADC PayloadsEdmund Graziani, Ph.D., Associate Research Fellow, Worldwide Medicinal Chemistry, Oncology East, Pfizer Global Research & DevelopmentNew potent natural product RNA splicing inhibitors have been enabled as payloads for antibody drug conjugates (ADCs). A number of different linker approaches were employed to address challenges associated with payload reactivity and stability. Similarly, both conventional (random) and site-specific conjugation methods have generated potent and efficacious ADCs that have a therapeutic index that is strongly correlated to drug antibody ratio (DAR).







4th Annual Bispecific Antibody Therapeutics

Engineering Multispecificity

JANUARY 22-23





























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Creating bioactive molecules that are multivalent and multifunctional offers the promise of more effective therapeutics. By binding to at least two molecular targets simultaneously, antibodies are empowered, thereby delivering a highly potent therapeutic, particularly for cancer immunotherapy. This Bispecific Antibody Therapeutics conference explores the challenges of engineering multispecificity to ensure stability and efficacy, and reviews the numerous forms of multispecific antibodies. The conference gives particular focus to safety issues, as well as PK and immunogenicity. Case studies highlight preclinical development as well as clinical data.





BISPECIFICS FOR ONCOLOGY

2:00 Chairperson’s Opening RemarksGunasekaran Kannan, Ph.D., Principal Scientist and Group Leader, Biologics Optimization, Amgen, Inc.


2:05 Cancer Immunotherapy with Bispecific T Cell Engager (BiTE®) AntibodiesDirk Nagorsen, M.D., Ph.D., Global Development Leader, Global Clinical Development, Amgen, Inc.T cells are an important component of cancer immunosurveillance. Various approaches have been tested to use T cells for the treatment of malignant diseases. Bispecific T cell Engager (BiTE®) antibodies represent an innovative investigational approach that is designed to engage the body’s endogenous T cells to target malignant cells. This talk will give an overview of the development status of BiTE® antibodies including the most advanced molecule in this class, blinatumomab, which targets CD19 on malignant cells of B lineage.

2:45 A Novel Fc-Containing Bispecific Format with Full-Length Antibody Properties: Applications in OncologyDavid Szymkowski, Ph.D., Senior Director, Biotherapeutics, Xencor, Inc.Bispecific antibody development has been hindered by inferior stability, production and half-life. We have engineered the Fc domain to create bispecifics possessing full-length antibody properties, including excellent stability and manufacturability, and extended half-life. Xencor’s “XmAb®” candidates exploit a common anti-CD3 domain that recruits T cells to potently kill malignant cells. I will discuss our development of bispecific antibodies targeting several tumor antigens, including demonstration of safety and efficacy in monkeys.

3:15 Enhanced Tumor Cell Killing via Selective Inhibition of CD47 with Bispecific AntibodiesNicolas Fischer, Ph.D., Head, Research, Novimmune SACD47 is a broadly expressed cell surface glycoprotein that is up regulated by tumour cells to evade surveillance by innate immune cells. We have generated and

evaluated a panel of CD47xCD19 dual-targeting bispecific antibodies selectively inhibiting CD47 on tumour cells. Based on in vitro and in vivo experiments we have selected a lead therapeutic candidate that mediates effective tumour killing and has an appropriate pharmacokinetic profile in non human primates.

3:45 A New Platform to Automate Bispecific Antibody Design and AssessmentChristopher Smith, Ph.D., Senior Scientific Consultant, Biologics, GenedataNext-generation antibodies, and specifically bispecifics, provide significant advantages over traditional IgG-based therapeutics. However, as highly engineered molecules they pose new design, cloning, expression, purification and analytics challenges. Our workflow platform fully automates the design, production and testing of large panels of multispecific antibody candidates, including tandem-scFv-Fc, KinH, DVD-Ig or diabodies. We demonstrate the platform’s handling of linkers, V-domains and Fc regions, its high-throughput capability and built-in tools for the systematic identification of development candidates based on a systematic analysis of assay, analytic and sequence data.


BISPECIFIC BREAKTHROUGHS5:00 MM-141, an IGF-IR- and ErbB3-Directed Bispecific Antibody, Overcomes Adaptations in Growth Factor Signaling NetworksAlexey A. Lugovskoy, Ph.D., Vice President, Therapeutics; MM-141 Project Leader, Merrimack Pharmaceuticals, Inc.Bispecific antibodies surpass the activity of conventional immunoglobulins and often possess added components of mechanisms of action. However, bispecifics have proven difficult to develop due to both suboptimal design and poor pharmaceutical properties. A Network Biology approach to drug design overcomes these complexities and accelerates the discovery of active and manufacturable therapeutic molecules. This approach will be illustrated by case studies of bispecific antibodies for the treatment of cancer developed at Merrimack Pharmaceuticals.

5:30 Therapeutic Fully Human Bispecific Antibodies with Two Binding Sites in Each Fv RegionKristian Jensen, Ph.D., Vice President, Project Management, R&D, Dutalys GmbHDutaMabs are fully human bispecific antibodies, comprising one normal heavy chain and one normal light chain, with two non-overlapping binding sites within the natural CDRs of each Fv. These two paratopes within the DutaMab CDRs are fully independent, due to an unprecedented stability, which exceeds that of all previously described antibodies. This allows the combination of any two specificities within one Fv, and their independent maturation to low pM affinities.







JANUARY 22-23





























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FRIDAY, JANUARY 23


PROGRESSIVE ENGINEERING

8:30 Chairperson’s RemarksDavid Szymkowski, Ph.D., Senior Director, Biotherapeutics, Xencor, Inc.

8:35 Nonclinical Development of Multi-Targeting Biotherapeutics: More Targets, More ComplexityRodney Prell, Ph.D., Senior Scientist, Toxocology, Genentech, Inc. – A Member of the Roche GroupMulti-targeting biotherapeutics offer several advantages over conventional monoclonal antibodies (mAbs). These include the potential for greater efficacy by modulating multiple pathways, preventing the development of resistance mechanisms, and introducing unique mechanisms of action. However, as the number of targets increases, so does the complexity of the nonclinical development program. This presentation will provide general considerations for the nonclinical development of multi-targeting biotherapeutics along with some case studies to highlight these points.

9:05 Engineering the Binding Mode of an Antibody to Cross the Blood-Brain BarrierAlain Tissot, Ph.D., Head, Immune Biology, Roche pRED Large Molecule Research

9:35 Engineering Bispecific Antibody through Electrostatic Steering Mechanism: Heavy Chain Heterodimerization and Light – Heavy Chain PairingGunasekaran Kannan, Ph.D., Principal Scientist and Group Leader, Biologics Optimization, Amgen, Inc.Bispecific antibodies are projected to have broad clinical applications. However, co-expression of multiple light and heavy chains using recombinant technologies often leads to contaminants and poses purification challenges. We have modified the Fc region and the light and heavy chain interface of the Fab with targeted mutations in order to generate bispecific antibodies. The engineered bispecific antibodies have desirable in vivo and in vitro stabilities similar to conventional IgGs.


11:00 Novel Approach to Hemophilia A Treatment: ACE910, an Asymmmetric Bispecific IgG to Coagulation Factors IXa and XYukiko Okuyama-Nishida, Ph.D., Sub Lifecycle Leader, Primary Lifecycle Management, Chugai Pharmaceutical Co., Ltd.ACE910 is a recombinant humanized anti-factors IXa and X asymmetric bispecific IgG that places these two factors into spatially appropriate positions and mimics a cofactor function of factor VIII (FVIII) for treating hemophilia A. It’s expected to be a long-acting and subcutaneously injectable agent. In this talk, the molecular design, manufacturing and some clinical data of ACE910 will be presented.

11:30 Homogeneous Bispecifics and ADCs by Disulfide Bridging Using Next-Generation MaleimidesJames Baker, Ph.D., Senior Lecturer, Chemistry, University College LondonA powerful and general chemical platform technology is described for the construction of homogeneous antibody conjugates by site-specifically targeting and bridging the interchain disulfide bonds. Application to the efficient generation of bispecifics from scFv and Fab fragments, and ADCs from full antibodies, will be reported.

12:00 pm A Novel and Generic Approach for the Generation of Monovalent Bispecific IgGYariv Mazor, Ph.D., Scientist, Antibody Discovery and Protein Engineering, Medimmune LLCMonovalent bispecific IgG antibodies that structurally resemble native human IgG are emerging as a new class of molecules. However, these molecules are quite difficult to engineer and manufacture. We have developed a robust and generic platform called DuetMab for the production of correctly assembled monovalent bispecific IgGs that maintains the structure and developability properties of natural IgG. A comprehensive biochemical, biophysical, and unique functional analysis will be described.

12:30 After the Development Candidate has been selected: Successful Transition into an IND-Enabling Toxicology Program

Sponsored by

Michael V. Templin, Ph.D., DABT, Technical Director, SNBL USAToxicology is a major milestone for transition from preclinical research to clinical studies (i.e., Phase I). Development of multi-targeting/bispecific antibodies involves a balance of addressing fundamental questions for all biologics, with those unique to each candidate. Oncology-centric targets, immunomodulation, cross-species activity, and other considerations are essential for a robust toxicology program, and the data collected during preclinical evaluations are the basis for establishing study designs and study objectives for a Toxicology Program.

1:00 Session Break


PLATFORM TECHNOLOGY

2:00 Chairperson’s RemarksNicolas Fischer, Ph.D., Head, Research, Novimmune SA






JANUARY 22-23

2:05 DuoBody® Platform: A Versatile Platform for Lead Discovery in the Final FormatAran F. Labrijn, Ph.D., Senior Scientist, Antibody Sciences, Genmab BVThe DuoBody® platform represents a novel and elegant post-production technology for the generation of stable bispecific antibodies. General strategies, and considerations, for bispecific antibody discovery will be discussed. The suitability of the DuoBody® platform for bispecific discovery approaches, showing the importance to do this in the final format - illustrated by surprising findings, will be shown in a case study selecting a lead cMetxEGFR bispecific antibody.

2:35 Preclinical Development of a Fully Human Bispecific Antibody PlatformEric Smith, Ph.D., Associate Director, Bispecifics, Regeneron Pharmaceuticals, Inc.This presentation will describe a platform technology to generate fully human bispecific antibodies through the combination of Fc modifications that allow selective protein A purification and VelocImmune® derived antibodies that utilize a single defined light chain. This technology has been used to generate a T cell redirecting bispecific and in vitro and in vivo efficacy data will be discussed, as well as findings from preclinical toxicology studies.

3:05 Structure-Based Design of Novel Bispecific PlatformsBrian Kuhlman, Ph.D., Professor, Biochemistry and Biophysics, University of North Carolina School of MedicineThe presentation will describe computational and rational design approaches to support the development of novel bispecific antibody platforms with high stability, multiple geometries and multiple valencies. Applications enabled by these Bispecific molecules will be described.

3:35 Preclinical Activity of MCLA-128, an ADCC Enhanced Human Bispecific IgG1 Antibody Targeting the HER2:HER3 HeterodimerMark Throsby, Ph.D., CSO, Merus B.V.Proprietary platform technology was applied to generate the Biclonics® MCLA-128; a human common light chain bispecific antibody targeting HER2 and HER3. MCLA-128 specifically and potently inhibits ligand dependent HER2:HER3 signaling resulting in suppression of tumor growth in vitro and in vivo. This novel full-length bispecific antibody, that features ADCC enhancement, will begin clinical evaluation in patients with HER2 positive tumors in 2015.

4:05 A Novel Glycoenginereed Bispecific Antibody Format for Targeted Inhibition of EGFR and IGF-1R Demonstrating Unique Molecular Properties Jürgen Schanzer, Ph.D., Principal Scientist, Discovery Oncology, Roche Diagnostics GmbHThe novel one-arm single chain Fab IgG bispecific antibody (XGFR) targeting IGF-1R and EGFR demonstrated potent signaling inhibition and enhanced antibody-dependent cellular cytotoxicity. XGFR has shown in vitro and in vivo anti-tumor activity in pancreatic, lung and colorectal mouse xenograft tumor models. In summary, rationale design can help to overcome low expression yields and impaired effector functions of bispecific antibodies.






























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PIPELINE 3: FORMULATION & STABILITY

7th Annual Optimizing Biologics Formulation Development

Case Studies of Problem Solving for Challenging Formulations, Novel Biotherapeutics and Packaging/Device Systems

JANUARY 19-20





























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Each year, the PepTalk Optimizing Biologics Formulation Development meeting brings together an international audience of analytical and formulation scientists from leading industry companies to hear solutions to the most significant challenges in their field. For 2015, the Optimizing Biologics Formulation Development conference focuses on the correlation of predictive stability studies with actual clinical results, formulating high concentration proteins, the interface of drug products with delivery and packaging systems and the management of ever-increasing amounts of analytical data in the formulation function.

SUNDAY, JANUARY 18




MONDAY, JANUARY 19


9:00 Chairperson’s Opening RemarksWei Wang, Ph.D., Research Fellow, Research and Development, Pfizer

»KEYNOTE PRESENTATIONS

9:10 NanoCrud: Roles of Nanoparticles in Aggregation Pathways, Adverse Immunogenicity and Quality Assessment of Therapeutic ProteinsJohn F. Carpenter, Ph.D., Professor, Pharmaceutical Sciences; Co-Director, Center for Pharmaceutical Biotechnology, University of Colorado Anschutz Medical CenterRecently, valuable new insights have been gained by characterizing nanoparticles in therapeutic protein products. For example, we found that nanoparticles present in solutions of intravenous immunoglobulin serve as precursors for microparticles during pharmaceutically relevant stresses (e.g., freeze-thawing or agitation). Also, therapeutic proteins can adsorb to foreign nanoparticles, and nanoparticles of therapeutic proteins can induce adverse immunogenicity. Regulatory agencies now view quantitation and sizing of nanoparticles as important for product quality assessment.

9:45 Strategies for Establishing a Formulation Function – A MacroGenics Case StudyTom Spitznagel, Ph.D., Vice President, Development, MacroGenics, Inc.This presentation will discuss some of the challenges in establishing a formulation department at a smaller company. Strategies for balancing risk tolerance, resourcing, outsourcing, and timing will be discussed and illustrated with case studies. In addition to more traditional formulation topics, examples will include ensuring adequate analytics, transferring fill/finish processes, and ensuring dosing strategies are properly selected and supported through compatibility studies.

10:20 Coffee Break

PREDICTIVE ANALYTICAL STUDIES IN FORMULATION DEVELOPMENT

10:45 Estimation of Shelf Life Based on Accelerated Stability DataWei Wang, Ph.D., Research Fellow, Research and Development, Pfizer

Product shelf life, a critical quality attribute, needs to be adequate to support worldwide commercialization. One key task during product development is to test, estimate, and optimize product stability. Stability studies are often conducted under accelerated conditions to collect data in a short time period. Such data, however, may not predict accurately the real-time shelf life. This presentation discusses the general principles, challenges, and options in shelf life estimation.

11:15 Roles of Thermal Ramping and Isothermal Analyses in Predictive Formulation StudiesLisa A. Kueltzo, Ph.D., Staff Scientist, Formulation Development, Vaccine Production Program Laboratory, National Institutes of HealthThermal ramping and accelerated high temperature analyses, such as DSC and DSF, have been a staple of formulation development for several decades. The variable ability of these methods to predict the real time stability of biological products has always been a known drawback to this approach, and has been of increasing interest in recent years. This talk will examine the comparative value of accelerated temperature methods with alternate isothermal methods. The benefits and disadvantages of the techniques will be reviewed, and the relative predictive ability for real-time stability will be examined.

11:45 Studies on Developability and Predictive Stability TechniquesDeniz Temel, Ph.D., Postdoctoral Researcher, Technical Development, Biogen IdecDevelopability assessment may play an extremely important role in assessment of biomolecule candidates’ behavior and prevention of their possible failure during preclinical and clinical development. Developability studies are integrated with and would bridge between the discovery/design and development/delivery. My research focuses on various measured and calculated properties of monoclonal antibodies that may then serve as predictors and indicators of suitability of these important biomolecules in wide range of concentrations.






JANUARY 19-20





























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12:15 pm Solid State Formulation of Therapeutic Antibodies Using Spray Drying for High Concentration Reconstitution

Sponsored by

A DIVISION OF DOSAGE FORM SOLUTIONS

Jeff Breit, Ph.D., Director, Bend Research The development of high concentration antibody solutions is being explored for use both in material logistics and storage as well as for clinical application. We will present a case study wherein we begin dissecting the variables important for the development of a spray-dried antibody formulation with a focus on drying kinetics, excipients and protein stability. Utilizing the information accrued in this program, we have developed a model for platform development of a spray-dried monoclonal antibody formulation.

12:45 Session Break

1:00 Luncheon Presentation I: Automated High-Throughput Approaches from Developability to Formulation Development of Biopharmaceuticals

Sponsored by

Russell Burge, Ph.D., Applications Scientist, Freeslate Inc.Automated high-throughput methods were applied to developability studies and early to late stage formulation development of biopharmaceuticals. Automation integrated with instrumentation generated high quality data and led to highly efficient studies in terms of sample and resource utilization. Results of the case studies highlight the utility and flexibility of laboratory automation and integration with analytical devices such as spectrophotometers, liquid chromatography (HPLC and UPLC) and dynamic light scattering DLS.

1:30 Luncheon Presentation II: Isothermal Chemical Denaturation in the Development of Protein PharmaceuticalsErnesto Freire, Ph.D., Professor, Biology and Biophysics, Johns Hopkins University Conformational stability, protein aggregation and solution viscosity are critical issues in the development of protein pharmaceuticals, especially high concentration formulations. Achieving optimal formulation conditions is a difficult balancing act, as excipients used to minimize aggregation or to lower viscosity often have an adverse effect on conformational stability, thus compromising long term stability. The linkage between stability, aggregation and viscosity can be assessed by measuring the free energy of stability (ΔG) and its concentration and time dependence. For monoclonal antibodies and other biologics that exhibit irreversible temperature denaturation, the only way of measuring ΔG is by Isothermal Chemical Denaturation (ICD). Having access to ΔG allows the implementation of more precise and faster formulation optimization algorithms. In this presentation, the fundamentals and applications of ICD to formulation optimization will be discussed.

OVERCOMING FORMULATION CHALLENGES

2:00 Chairperson’s RemarksMurali Bilikallahalli, Ph.D., Associate Director, Formulation Sciences, Proteins, Vaccines & Oligos, MedImmune

2:05 Formulation Development for Highly Unstable ProteinsRamil Latypov, Ph.D., Associate Director, Genzyme

2:35 Pharmaceutical Challenges during Late Stage Development of an ADCNia Adeniji, Senior Research Associate, Genentech, Inc.

3:05 Case Study: Challenges in Developing Multi-Protein/Antigen Drug ProductsMurali Bilikallahalli, Ph.D., Associate Director, Formulation Sciences, Proteins, Vaccines & Oligos, MedImmuneCo-formulated drug products containing two or more therapeutic proteins or engineered vaccine antigens are challenging to develop. But these give significant advantage over mixing of multiple drug products at the bedside in terms of cost of manufacturing and patience convenience. This case study describes some of the formulation and analytical challenges including developability assessment, multi-protein compatibility and overlapping degradation pathways.

3:35 Selected Poster Presentation: Case Study: Early Assessment Strategies for Comprehensive Acceleration of Biologic TimelinesLori B. Karpes, Ph.D., Scientist, Protein Formulation Development, Biogen Idec


4:15 Case Study: Handling the Data Deluge in Formulation DevelopmentSteven LaBrenz, Ph.D., Scientific Director, Drug Product Development, Janssen R&DThe adoption of multivariate analysis during formulation development as a requirement QbD may necessitate the use of HTS. The generation of data sets that as a result of HTS are very large and complex quickly limits the productivity gains associated with these techniques. A “small data” approach where data is held in a local database and turned into discreet pieces of information delivers value to laboratory personnel by increasing local productivity.

4:45 Antimicrobial Preservatives in Parenteral Formulations – Practical Implications of Peptide/Preservative InteractionsPetteri Heljo, Ph.D., Postdoctoral Researcher, Early Stage Pharmaceutical Development, F. Hoffmann-La RocheAntimicrobial preservatives such as benzyl alcohol, phenol or m-cresol must be added to parenteral multidose formulations to ensure sterility maintenance during repeated product administration, i.e. multi-use of a parenteral drug product. However, preservatives may interact with API or other excipients, possibly altering their solution properties and physicochemical stability. This presentation explores the relevance of these phenomena in parenterally administered peptide formulations from the perspective of API stability and microbial viability.






JANUARY 19-20





























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5:15 Fluorescence and Light Scattering Methodologies in Biopharmaceutical DevelopmentMalgorzata Tracka, Scientist II, Formulation Sciences, MedImmunePlatform technologies and approaches are typically utilised by the industry during the bioprocessing of monoclonal antibodies (mAbs). However, recent advances in protein engineering have generated novel ‘non-mAb formats’, which pose new challenges to the formulation scientist. These include new scaffold architectures with associated changes to pI, MW, conformation and colloidal properties and possible self-assembly. Since aberrant protein characteristics have the potential to adversely impact drug product quality, activity, immunogenicity and manufacturability, there is a need for new screening technologies that inform us of the biophysical state of non-mAb formats in a high throughput manner. Using case studies, we will outline new screening assays based on light scattering technologies and fluorescence, enabling us to define the key formulation parameters for early cycle development.


TUESDAY, JANUARY 20


FORMULATION IMPACTS OF PACKAGING AND DELIVERY SYSTEMS

8:30 Chairperson’s RemarksSwaroopa Paratkar, Ph.D., Research Investigator, Bristol-Myers Squibb

8:35 PFS Filling of High-Concentration (HC) mAb Formulations – Minimization of Formulation Drying and Nozzle CloggingYuh-Fun Maa, Ph.D., Principal Engineer, Pharmaceutical Processing & Technology Development, Genentech, Inc. – A Member of the Roche Group Syringe filling of high-concentration (HC)/viscosity mAb formulations is rarely published. The purpose of this study is three-fold: 1) To reveal design details of a bench-top syringe filling unit 2) To understand critical filling parameters 3) To apply the learning to minimize formulation drying leading to fill nozzle clogging. The outcomes of this study will benefit scientists and engineers who develop pre-filled syringe products by providing a better understanding of HC formulation filling principles and challenges.

9:05 Stability of Biopharmaceuticals in Plastic Pre-Filled Syringes in Comparison with Glass Pre-Filled SyringesSwaroopa Paratkar, Ph.D., Research Investigator, Bristol-Myers Squibb

9:35 Selected Poster Presentation: Size And Molecular Flexibility of Sugars Determine the Storage Stability of Freeze-Dried ProteinsMaarten Mensink, Ph.D. Candidate, University of Groningen


11:00 Development of Stable Oral Thin Film Formulations for Live Rotavirus VaccinesPhillip Lovalenti, Ph.D., Director, R&D, Aridis Pharmaceuticals

11:30 pm Multimode Stability Measurements in Formulation Screening and Characterisation of Biologics

Sponsored by

Geoff Platt, Ph.D., Head, Applications, Avacta Analytical, UKOptim delivers high throughput measurements of proteins which provide information on both their conformational stability and propensity to aggregate. It has been used to screen biologics formulations and its capacity to afford predictions of long-term stability for systems that degrade by different mechanisms has been demonstrated. The technology can aid optimisation and development of biologics by characterising degradation mechanisms of monoclonal antibodies, antibody-drug conjugates and membrane proteins, and has the ability to screen protein-ligand interactions.

12:00 Session Break

12:00 Enjoy Lunch on Your Own

2:00 BuzZ Session A









8th Annual Lyophilization and Emerging Drying Technologies

Formulation Development, Process Optimization, Validation and Regulatory Compliance

JANUARY 21-22





























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This popular conference covers latest trends and challenges in lyophilization, spray drying, foam drying and emerging drying technologies. The Lyophilization and Emerging Drying Technologies conference features in-depth case studies and discussion on developing scientifically sound formulation, process optimization for biologics and vaccines. It also presents cutting-edge research and case studies on stability challenges of lyophilized formulations, drying in cartridges, storage stability and strategies for scale-up from R&D scale to full production level, and advances in lyophilization and alternate drying technologies.

TUESDAY, JANUARY 20


2:00 BuzZ Session A







LYOPHILIZED FORMULATION DEVELOPMENT AND PROTEIN STABILITY

8:15 Chairperson’s Opening RemarksGerhard Winter, Ph.D., Professor, Chair, Pharmaceutical Technology and Biopharmaceutics, LMU Munchen


8:20 Neglected Factors in Stabilization of Proteins by Freeze DryingMichael Pikal, Ph.D., Distinguished Endowed Chair in Pharmaceutical Technology & Professor of Pharmaceutics, University of ConnecticutStability trends have traditionally been interpreted in terms of either native structure retention during the process or minimizing molecular mobility. While recent evidence does suggest that storage stability often correlates well with measures of “fast dynamics”, such as the amplitude of motion on a nanosecond time scale, correlations are not perfect. Here, we discuss two other factors that are often ignored, surface effects and coupling of the protein internal dynamics with dynamics of the matrix.

9:00 Lyophilization Process Design and Development Using Quality by Design (QbD) PrinciplesSajal M. Patel, Ph.D., Scientist, Formulation Sciences, Biopharmaceutical Development, MedImmune, Inc.Lyophilization is a unit operation that is routinely used to stabilize an otherwise unstable molecule to achieve pharmaceutically acceptable shelf life. The lyophilization process is based on fundamental principles of heat and mass transfer, and the overall understanding of the impact of process parameters on product quality attributes has increased significantly over the past few decades. QbD aims at building quality within the process rather than monitoring offline at the end of the process. This presentation will describe application of QbD elements—risk assessment, process characterization, PAT (Process Analytical Tool) and Design Space—to design, develop and scale-up of the lyophilization process.


ADVANCES IN LYOPHILIZATION TECHNOLOGY

10:50 In silico Analysis Design of Freeze-Drying Systems and ProcessesAlina A. Alexeenko, Ph.D., Assistant Professor, School of Aeronautics and Astronautics, Purdue UniversityThis talk reviews recent advances in application of in silico methods for understanding and improving lyophilization. Computational simulations have been applied to vial- and batch-level heat and mass transfer as well as modeling of various elements of lyophilizers such as shelf uniformity, product chamber flows and condenser performance. The integration of such models into a system framework provides a new process analytical tool for more efficient and robust freeze-drying of bio-pharmaceuticals.

11:20 A Critical Eye on Controlled Nucleation in Freeze Drying of Protein DrugsGerhard Winter, Ph.D., Professor, Chair, Pharmaceutical Technology and Biopharmaceutics, LMU MunchenControlled nucleation (CN) has gained attention as a process step promising reduced drying time and improved batch homogeneity. Many vendors offer systems that can be attached to existing freeze dryers. The question remains how much positive effect on the product (protein) quality can really be achieved by CN, whether it is valuable for highly concentrated protein formulations and how its implementation from formulations research into production should be managed.






JANUARY 21-22





























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11:50 Application of Controlled Ice Nucleation for Lyophilization of a Low Glass Transition Temperature SolutionGraham Magill, Engineer I, Pharmaceutical Processing and Technology Development, Genentech, Inc. – A Member of the Roche GroupHere we will discuss the application of Praxair ControLyo controlled ice nucleation technology to optimize the lyophilization of a low glass transition temperature (Tg’), high fill volume product. The impact of processing parameters such as ice nucleation temperature and post-nucleation cooling ramp rate on cake microstructure will be discussed.

12:20 pm Enjoy Lunch on Your Own

LYO PROCESS DEVELOPMENT: QbD, SCALE-UP AND DESIGN SPACE

2:00 Chairperson’s RemarksIan Whitehall, Vice President, Sales & Service, SP Scientific

2:05 Measurement of Vial Heat Transfer Coefficients: Nuances and PitfallsRobin Bogner, Ph.D., Associate Professor, School of Pharmacy, University of ConnecticutThe design of a lyophilization cycle, its scale-up, and several PAT tools require input of a “vial heat transfer coefficient”, Kv. The classic equations for measurement of Kv (described in 1984) are applicable to a well-defined experimental protocol. Modification of the Kv measurement protocol requires revised equations to accurately calculate Kv. The use of accurate vs. inaccurate Kv in lyophilization design space and PAT will be compared.

2:35 Primary Drying Optimization Using a Three-Dimensional Design SpaceArnab Ganguly, Ph.D., Research Assistant, School of Aeronautics and Astronautics, Purdue University Lindsay A. Wegiel, Ph.D., Research Associate III, Pharmaceutical R&D Baxter Healthcare CorporationThe current approach to primary drying optimization using a design space is two dimensional whereby the processing conditions are set throughout the primary drying cycle. However, the product is changing throughout primary drying with regards to the product resistance (Rp). This is a key component of the design space calculations and thus adding the third dimension of time to allow for the changes in Rp will allow for further optimization and shorter lyophilization cycle times.

3:05 Laboratory Measurement of Collapse Temperature Predictive of Collapse in Manufacturing OperationsVamsi Mudhivarthi, Ph.D., Postdoctoral Fellow, School of Pharmacy, University of ConnecticutTraditionally, freeze drying microscopy has been used to estimate collapse temperature, but it has become apparent that such methodology is not necessarily predictive of collapse during freeze drying in commercial containers. Previous studies suggested that methodology based on “Optical Coherent Tomography”, or OCT, is predictive. Here we demonstrate the generality of this early observation and present a theoretical analysis that supports the experimental observations using OCT.

3:35 Freeze Drying PAT using Heat Flux Measurement Sponsored by

T.N. Thompson, President, Millrock Technology, Inc A freeze drying recipe will be analyzed and then improved using Heat Flux measurement and Control. Heat flux measurements provide critical process information for every step of the freeze drying process, including: freezing, primary drying and secondary drying. By measuring heat flux in-process process parameters such as % of product frozen by nucleation, crystal growth rates post-nucleation(Controlled and UnControlled), end of freezing, thermal conductivity of the vial (Kv), product resistance (Rp), mass-flow, shelf surface temperature and product temperature can be measured. Additionally, a design space can be developed in a single run.






ADVANCES IN ALTERNATE DRYING TECHNOLOGIES

8:30 Chairperson’s RemarksSajal M. Patel, Ph.D., Scientist, Formulation Sciences, Biopharmaceutical Development, MedImmune, Inc.






JANUARY 21-22





























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8:35 Use of Microwave Vacuum Drying as an Alternative to Lyophilization of Vaccines and BiologicsJustin Stanbro, Ph.D., Associate Principal Scientist, Novel Adjuvants, Formulation & Delivery Technologies, Merck & Co.Lyophilization is an established unit operation for the dehydration of sensitive vaccine and biologic products that greatly improves a product’s thermostability profile. However, long cycle times often due to mass and heat transfer limitations along with limited formulation space frequently present bottlenecks in drug development efforts. Here we present the use of microwave vacuum drying as an alternative to lyophilization capable of drying in 4-8 hours while maintaining critical product attributes and drying uniformity.

9:05 Advances in Foam Drying Formulation and Process Technology for BiologicsDavid Thomas, Principal Scientist, Bio Therapeutics R&D, Pfizer, Inc.Foam drying is an alternative drying technique that has potential as an enabling technology for freeze sensitive biologics. New process capabilities along with novel formulation approaches are described. Additional applications with high concentration proteins are also discussed. Foam drying is an underutilized and poorly understood technique that has a number of potential advantages over freeze drying, including reduction in reconstitution time and enabling of dried vaccine formulations.

9:35 Selected Poster Presentation: Investigating the Impact of Solute and Fill Volume on Vial Heat Transfer CoefficientsPooja Sane, Ph.D. Candidate, Pharmaceutics Department, University of ConnecticutVial heat transfer coefficient (Kv) is one of the key determining factors of product temperature at any processing condition during freeze-drying. Conventionally, Kv has been determined by water sublimation experiments with a fixed fill volume. In this study, we have explored the use of water versus representative products at different fill volumes in estimation of the batch average Kv and variation across the batch. This study is an attempt to improve the uniformity of product quality across a shelf and for efficient scale-up to a manufacturing scale dryer using real product.


LYOPHILIZATION CHALLENGES: DUAL CHAMBER CONTAINERS, SYRINGES AND GLASS VIALS

10:50 Lyophilized Drug Product Fogging onto the Glass Vial: Problem and SolutionMark Yang, Ph.D., Director, Fill Finish Development, Commercial Process Development, Genzyme, a Sanofi CompanyA case study is presented where a biological product creeps up the glass vials and forms amorphous a thin while film (fogging) above the cake after lyophilization. Several mitigation strategies have been investigated, including lyophilization cycle optimization, surfactant and formulation adjustment, filling temperature, vial sourcing, and vials type screening. It is found that a commercial vial with new surface property can completely eliminate this fogging problem.

11:20 Development of Lyophilized Protein Product in Dual Chamber SyringeBingquan (Stuart) Wang, Ph.D., Senior Scientist, Biogen IdecThe challenges in the formulation and process development in Dual Chamber Syringe (DCS) will be discussed and compared to a typical vial container. A case study is presented in order to improve the batch uniformity during scale up. The heat transfer mechanism in DCS is investigated in different holder systems, and the mass transfer in DCS is also studied and compared to that in a vial. The characterization of heat and mass transfer in DCS is critical to develop a robust lyophilization process.


11:50 Lyophilization of Pharmaceuticals in Dual Chamber Containers (DCC): Challenges during Process Scale-Up (Case Studies)Serguei Tchessalov, Ph.D., Associate Research Fellow, Biotherapeutics Pharmaceutical Research & Development, Pfizer, Inc.The specifics of heat and mass transfer in DCC during freeze-drying of pharmaceuticals will be discussed with a focus on the evolution of magazines (DCC supporting systems). The case studies will demonstrate the difficulties in prediction of the scale up issues for both semi-crystalline and amorphous materials. The approaches to the rational lyo robustness studies, with respect to process design space, would also be presented.







6th Annual Protein Aggregation and Emerging Analytical Tools

Mechanism, Prediction, Screening, Immunogenicity and Formulation Challenges

JANUARY 22-23





























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The popular Protein Aggregation and Emerging Analytical Tools conference covers latest trends, challenges and solutions in protein aggregation. It features in-depth case studies and interactive discussions on mechanisms of aggregation, detection and quantitation of aggregates and rapid analytical techniques. It also presents case studies on prevention of particle formation by engineering and formulation approaches, impact of aggregation on production, aggregates as a factor for immunogenicity and approaches for improvement of biophysical properties of protein solutions.





UNDERSTANDING PROTEIN INTERACTIONS FOR BETTER PRODUCT DESIGN AND DEVELOPMENT

2:00 Chairperson’s Opening RemarksAnette Henriksen, Ph.D., Principal Scientist, Protein Biophysics and Formulation, Novo Nordisk


2:05 Protein-Solvent Interaction and Its Importance to Solubility and ViscosityThomas Laue, Ph.D., Professor, Biochemistry and Molecular Biology; Director, Biomolecular Interaction Technologies Center (BITC), University of New HampshireProteins interact with the surrounding solvent in both specific and non-specific ways. The hydration shell must be displaced for protein-protein interactions to occur. Also, proteins bind monovalent ions, particularly anions, which will affect protein-protein interactions. Previous work provides insights into how these protein-solvent interactions may impact protein solubility and viscosity. These insights are useful for guiding protein design and solvent selection.

2:45 Aggregation of Antibody CH2 Domains and Antibody-Drug ConjugatesDimiter S. Dimitrov, Ph.D., Senior Investigator, Protein Interaction Group, FNLCR, NCI, National Institute of HealthAggregation of isolated antibody domains will be overviewed and our recent work on CH2 and updates will be discussed (Gong R at al Mol Pharm. 2013, unpublished). Aggregation of antibody-drug conjugates (ADCs) will be discussed. Experimental data and computational models will be presented for some ADCs including those generated in our group.

3:15 Stable Human IgG Therapeutics through Engineering of Antibody Variable DomainsDaniel Christ, Ph.D., Head Antibody Therapeutics, Immunology, Garvan Institute of Medical ResearchHuman antibody variable domains (VH and VL) that mediate the interaction with antigen often display poor stability and a propensity to aggregate. We have recently developed generally applicable phage display and X-ray crystallography strategies that allow the stability engineering of human variable domains. Here we outline the application of the technology to full-length human IgG, and present examples of how the approach can be utilised for the ‘retrofitting’ of candidate molecules and biobetters.

3:45 Sub-micron, Sub-mL Particle Counting and Sizing with Focused Beam SPOS Technology: Ideal for Protein Aggregate and Small-Volume USP 787 Analysis

Sponsored by

David F. Nicoli, Ph.D., Vice President, Research & Development, Particle Sizing Systems LLCA new single-particle optical sizing (SPOS) technique uses a focused laser beam to count and size protein aggregates from 0.15 to 20 microns and at high concentration, inaccessible by conventional light obscuration and scattering. Addition of a combined light obscuration/scattering sensor extends the upper size limit to 150 microns. Analysis can be made on sub-mL volumes, with sample conservation. Samples of high viscosity resulting from high protein concentration (> 100 mg/mL) can be measured without difficulty.


CHARACTERIZATION OF PROTEIN AGGREGATES

5:00 Detection and Characterization of Visible, Subvisible Particles and Other Aggregates: Achievements and ChallengesAnacelia Rios Quiroz, MSc, Assistant Scientist, Late-Stage Pharmaceutical and Process Development, Pharmaceutical Development & Supplies, PTD Biologics Europe, F. Hoffmann-La Roche Ltd.The talk will give an overview of required and commercially available counting methodologies for detection of protein aggregates and visible and sub visible particles (SbVP); species ubiquitously present in protein formulations. Focus will be SbVP as they are gaining attention regarding immunogenicity and quality attributes. Lack of a well-defined methodology for SbVP makes important to increase our knowledge of emerging instruments’ performance. Applicability towards the assessment of a meaningful array of particle counting techniques will be discussed.






JANUARY 22-23





























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5:30 Characterization of Protein Aggregate Structure with SAXSAnette Henriksen, Ph.D., Principal Scientist, Protein Biophysics and Formulation, Novo NordiskProteins can aggregate through a number of different mechanisms, possibly leading to different structure and biological activity of the aggregates. We have investigated the structure of a mAb dimer by Small Angle X-ray Scattering coupled to Size Exclusion Chromatography. The results clearly show that the internal structure of the monomer is conserved to a high degree, and furthermore provide information on the relative orientation of the monomers within the dimer.


FRIDAY, JANUARY 23


PROTEIN AGGREGATION AND PRODUCT STABILITY: PREDICTION, RAPID SCREENING AND NEW TOOLS

8:30 Chairperson’s RemarksDaniel Christ, Ph.D., Head Antibody Therapeutics, Immunology, Garvan Institute of Medical Research


8:35 Microscale-Thermophoresis: A New Tool for High-Throughput Protein Stability Prediction?Gerhard Winter, Ph.D., Professor, Chair, Pharmaceutical Technology and Biopharmaceutics, LMU MunchenMicroscale-Thermophoresis (MST) has been developed a few years ago to determine label free protein-ligand binding with a novel laser based analytical concept. Only few µl sample volume is required and both substrates are in solution. We now study how the method could serve as an alternative to µDSC and HTSF to determine protein melting curves in different pharmaceutical formulations. Data on different model proteins including monoclonal antibodies are presented.

9:05 Probing the Interrelationships between Protein Higher-Order Structure, Formulation Composition, and Pharmaceutical StabilityDavid B. Volkin, Ph.D., Distinguished Professor, Department of Pharmaceutical Chemistry, University of KansasThis talk will cover new analytical approaches to assess protein physical stability profiles as part of formulation development and comparability assessments. Case studies will include (1) A novel chaperone protein biosensor (GroEL-BLI) to probe for preaggregate species in stressed protein solutions (2) High-throughput biophysical analyses combined with data visualization tools examining various IgG1-Fc glycoforms and (3) H/D exchange mass spectrometry to correlate local flexibility and physical stability changes for IgG1-mAb with different additives.

9:35 Approaches to Separately Predict Aggregation Mechanisms and Shelf LifeChristopher J. Roberts, Associate Professor, Department of Chemical & Biomolecular Engineering, University of DelawareThis presentation first focuses on combining in situ biophysical techniques to measure protein-protein and protein-solute interactions as a means to predict how aggregation mechanisms change with formulation conditions. Independent of one’s knowledge of aggregation mechanism(s), a separate technique allows one to more accurately estimate shelf life and confidence intervals from accelerated data.


11:00 The Simultaneous Determination of the Extent and Mechanism of Aggregation and the Stability of Proteins as Key Factors to be Considered in DevelopabilityBelinda Pastrana, Ph.D., Full Professor, Chemistry, University of Puerto RicoThe development of novel protein therapeutics can take advantage of the determination of the mechanism and extent of aggregate species under different formulations conditions. We have developed an innovative, accurate and reproducible spectroscopic approach to ascertain this vital information. This method also allows for the determination of stability of the domains of the full-length protein. Two case studies will be presented: (1) IgG2a and (2) Related calcium binding proteins.

11:30 An Improved Procedure for Quantitating Size-Related Degradation Products by Sedimentation Velocity Analytical UltracentrifugationYatin R. Gokarn, Ph.D., Drug Product and Device, Biologics Development, Gilead Sciences, Inc.This presentation will discuss an improved procedure for quantitating size related degradation products in biopharmaceutical by using sedimentation velocity analytical ultracentrifugation.

12:00 pm Fill-Finish Process Challenges: Case Study of a High Concentration Monoclonal AntibodyChinmay Bakshi, Associate Scientist, Formulation Sciences, MedImmune, Inc.Filter and dispense process steps are critical components of the drug product (DP) manufacturing process for ensuring the safety and efficacy of the final dosage form. Optimizing the multitude of fluid-handling process parameters is of utmost importance given the high viscosity and aggregation propensity of high concentration protein therapeutics. Development studies supporting component selection (pump, tubing, filter, orifice and nozzle diameter), commercial scale challenges including nozzle clogging and verification of the scale-down model will be covered.






JANUARY 22-23





























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12:30 Selected Poster Presentation: Characterization of Post-Injection Interactions between Monoclonal Antibodies in Subcutaneous Tissue and SerumErin Green-Trexler, Scientist, Vaccine Bioprocess Research and Development, Merck Research Laboratories, Merck Sharp & Dohme Corp.As the market for monoclonal antibodies expands, understanding the factors influencing efficacy and safety following injection becomes increasingly important. We have explored two aspects of post-injection phenomena between monoclonal antibody drug products and surrounding tissues by utilizing in-vitro studies. Factors influencing bioavailability were examined by analyzing concentrated mAbs following exposure to rat subcutaneous tissue. Additionally, an SPR method was developed to evaluate the possibility of interaction and aggregation of mAb products with endogenous serum proteins.

1:00 Session Break


ENSURING STABILITY, SAFETY AND EFFICACY OF BIOPHARMACEUTICALS

2:00 Chairperson’s RemarksMarisa K. Joubert, Ph.D., Senior Scientist, Process & Product Development, Amgen, Inc.


2:05 Protein Aggregate Size: A Potentially Important Factor in Inducing an Immune Response in Both in vitro and in vivo Model SystemsMarisa K. Joubert, Ph.D., Senior Scientist, Process & Product Development, Amgen, Inc.This presentation will discuss the impact of size of protein aggregates and will discuss in detail how it can play a potentially important role in inducing an immune response in both in vitro and in vivo model systems.

2:35 Examining the Leachable from Disposable Bioprocess Bag and Their Impact on Protein StabilityNina Xiao, Ph.D., Senior Research Associate, Late Stage Pharmaceutical Development, Genentech, Inc.Leachables from BioProcess Containers (BPCs) are source of process-related impurities that have the potential to alter product quality and affect patient health. In our studies, we observed IgG1 instability after storage in gamma irradiated single-use plastic BPCs. Analysis of protein’s quality attributes provided useful hints on potentially unidentified leachables induced by gamma irradiation of BPCs. Risk mitigation for preventing the effect of leachables on protein instability is discussed.

3:05 Surface Plasmon Resonance imaging (SPRi): A Multifaceted Tool for Protein Analysis in SerumMarinella Sandros, Ph.D., Assistant Professor, Department of Nanoscience, University of North Carolina at GreensboroSPRi is a label-free surface - sensitive optical detection method for monitoring biomolecular interactions in real time with high-throughput. Attendees will gain a better understanding of how SPRi can be used as a flexible tool to screen and study protein interactions and to assess stability in a complex environment. Since SPRi detection is a non-destructive method, retrieved proteins can be analyzed simultaneously with MALDI - TOF, fluorescence and electrochemistry.

3:35 Extractables/Leachables in Biopharmaceutical Manufacturing Processes: Sources, Risks, and Case StudiesKathryn McGohan,M.S., Associate Scientist II, Manufacturing Sciences & Technology, Bristol-Myers Squibb Co.Extractables/leachables pose many challenges and concerns in biopharmaceutical manufacturing processes. These challenges and concerns are significantly increased when single-use products are used. Successful management of extractables/leachables issues includes identifying the sources of the compounds, understanding the process requirements, and analyzing the potential risks. This talk will present several case studies where different strategies were used by cross-functional teams to identify and control the risks related to material extractables/leachables.




PIPELINE 4: EXPRESSION & PRODUCTION

7th Annual Engineering Genes, Vectors, Constructs and Clones

Upstream Decisions Lead to Downstream Success

JANUARY 19-20





























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As demand for high-quality biotherapeutic proteins grows, bottlenecks arise because functional recombinant proteins are difficult to produce. Protein expression engineers must often design new cloning schemes, including lengthy verification and gene or protein sequence analysis, move genes across vectors, transfect vectors in alternative hosts, re-characterize expressed proteins or any of the above—an inefficient, time-consuming and expensive process. The Engineering Genes, Vectors, Constructs and Clones conference returns to showcase effective engineering strategies for protein expression and production research leading to functional biotherapeutic products.

SUNDAY, JANUARY 18




MONDAY, JANUARY 19


SYNTHETIC BIOLOGY

9:00 Chairperson’s Opening RemarksMark Welch, Ph.D., Director, Gene Design, DNA2.0, Inc.


9:10 Design, Build, Test and Learn Strategies for Genome EngineeringRyan T. Gill, Ph.D., Associate Professor, Chemical and Biological Engineering, University of Colorado and Associate Director and Fellow, Renewable and Sustainable Energy InstituteAdvances in DNA synthesis and sequencing are enabling new approaches for engineering biological systems. Such approaches build off of decades of broader efforts in the engineering and computational sciences to direct the design of proteins, pathways and organisms to perform desired functions. This presentation describes such approaches and how they are being applied to improved production of various target molecules.

9:50 The Engineering of Artificial Cellular Nanosystems Using Synthetic Biology ApproachesCheemeng Tan, Ph.D., Assistant Professor, Biomedical Engineering, University of California, DavisArtificial biological systems represent effective platforms to test synthetic components beyond cells, to produce target molecules in vitro and enable new therapeutic approaches. I discuss two popular ones. Cell-free systems are isolated circuits assembled with biological components to perform defined tasks in vitro. But artificial cells are biomimetic and compartmentalized compositions of transcription-translation machinery combined with biological information. These systems are widely relevant for producing bio-commodities and biomedical applications.

10:20 Coffee Break

10:45 An Integrative Genome-Scale E. coli Model for Targeted Experimentation in Systems and Synthetic BiologyIlias Tagkopoulos, Ph.D., Assistant Professor, Computer Science and Genome Center, University of California, DavisWe present an integrative Escherichia coli model that bridges the transcriptional, signal transduction and metabolic layers through a constraint-based framework. Training of this modeling over the most extensive normalized genome-scale compendium resulted in a performance highly predictive of growth and expression under various environments. We discuss how such models can be used in synthetic biology in conjunction with automated design platforms and new extensions for integration of proteomics and condition-specific predictions.

11:15 Mammalian Systems and Synthetic Biotechnology for Recombinant Protein ProductionDong-Yup Lee, Ph.D., Assistant Professor, Chemical and Biomolecular Engineering, National University of Singapore; Senior Research Scientist, Bioprocessing Technology Institute, A*STARThe increasing demand for recombinant therapeutic proteins highlights the need to constantly improve the efficiency, yield and quality of these products from mammalian cells. We have recently established a “mammalian systems and synthetic biotechnology” framework where omics-data-driven and hypothetical model-driven approaches are integrated to study their growth characteristics, enhance cellular performance and design novel biological products or functions. This talk presents several applications of the framework.

11:45 Expansion of the Genetic AlphabetFloyd Romesberg, Ph.D., Associate Professor, Chemistry, The Scripps Research InstituteWe have generated an expanded genetic alphabet consisting of the two natural base pairs, A–T and G–C, and a third unnatural base pair of our own design. Recently, we have demonstrated that Escherichia coli may be engineered to import the requisite unnatural triphosphates, and that the resulting six-letter DNA is efficiently replicated and transcribed within the cell. This represents the first semi-synthetic organism that can store and retrieve increased information in its genome.






JANUARY 19-20





























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12:15 pm Robust Expression and Purification of Correctly Folded and Functional Ion ChannelsYelena Bisharyan, Ph.D., Senior Scientist, Tetragenetics, Inc. We have developed a technology that harnesses the naturally abundant ion channel producing capabilities of Tetrahymena thermophila to enable high-density display of correctly folded and functional recombinant human ion channels on the Tetrahymena cell surface. A drug discovery toolbox has been developed that includes surface membrane fractions enriched in these channels and large (milligram) amounts of functionally reconstituted purified ion channels for use as immunogens for antibody discovery and/or incorporation into platforms for small molecule drug screening.

12:45 Session Break

1:00 Luncheon Presentation: Systematic Sponsored by Engineering of Biological SystemsMark Welch, Ph.D., Director, Gene Design, DNA2.0, Inc.Current gene synthesis technologies allow unprecedented capability to tailor biological systems for a wide range of purposes. We describe how gene synthesis in concert with machine learning methods can be used to identify and engineer gene, pathway, genome or protein sequence variables critical for performance. Examples of application for protein expression, pathway optimization and protein engineering are discussed.

ANALYZING CONSTRUCTS, OPTIMIZING CODONS, AND ENABLING VECTORS

2:30 Chairperson’s RemarksJoseph D. Kittle, Jr., Ph.D., Assistant Professor, Chemistry and Biochemistry, Ohio University; Founder and CSO, MTL, LLC

2:35 A Scoring System for Predicting Solubility of Protein ConstructsBrian Chiswell, Ph.D., Assistant Professor, Touro College; Owner and Founder, Dimensions BioSciencesThe amino acids chosen to be included in a protein expression construct can greatly affect the usability of a protein in the laboratory. Dimensions BioSciences has developed a scoring system that systemically analyzes the primary and secondary structure of a construct and determines the amino acid sequence most likely to remain soluble throughout purification and downstream experiments. Current applications and calibrations for our scoring system will be presented.

3:05 A Dual Host Vector for Fab Phage Display and Expression of Multiple Antibody Formats in Mammalian CellsIsidro Hötzel, Ph.D., Senior Scientist, Antibody Engineering, Genentech, Inc.Phage display is widely used in discovery of therapeutic antibodies. A bottleneck in the screening of clones from phage display libraries is the subcloning of variable regions to mammalian vectors for expression of IgG. We developed a vector for Fab phage display that expresses multiple antibody

formats in mammalian cells without reformatting, expediting the screening of clones derived by phage display.

3:35 Strep-tag/Strep-Tactin System – Versatile Toolbox for Recombinant Proteins

Sponsored by

Thomas Schmidt, Ph.D., COO, IBA GmbHThe Strep-tag®/Strep-Tactin® system is commonly known to provide highly pure and functional proteins at reasonable cost. IBA´s latest development, Strep-Tactin XT, completes now the product portfolio by enabling stable but still reversible immobilization for novel assay applications


4:15 Quantification of Cytosolic Plasmid DNA Degradation Using High-Throughput Sequencing: Implications for Gene DeliveryMark M. Banaszak Holl, Ph.D., Professor, Chemistry and Biomedical Engineering, University of MichiganCytosolic DNA degradation plays an important role in decreasing transgene expression; however, the cleavage locations remain largely unexplored. High-throughput sequence mapping of cytosolic nuclease cleavage sites for Luciferase plasmid in HeLa cells revealed the following most common cut sites: the poly(A) region between the β-lactamase gene and the cytomegalovirus promoter, the 5’ end of the β-lactamase gene, the OriC region, the SV40/poly(A) region, the luciferase gene and the CMV promoter.

4:45 Recombinant Protein Expression and Engineering: Generation of Expression Vectors and Protein BindersAgustín Correa, Ph.D., Research Scientist, Recombinant Protein Unit, Institut Pasteur de MontevideoRecombinant protein expression is a valuable tool in many industrial applications; however, expression problems often arise. We have generated a vector suite for easy cloning and evaluation of different expression parameters affecting recombinant protein production yields. An attractive application for some recombinant proteins is their use as binding reagents. We evaluated a new library design derived from Sac7d variants (Affitins) to generate binding proteins that can work as potent protein inhibitors.

5:15 Thinking beyond the Plasmid: Rapid Engineering of Stable Protein Expression and Improved Bacterial Strains Using Chromosomal EngineeringJoseph D. Kittle, Jr., Ph.D., Assistant Professor, Chemistry and Biochemistry, Ohio University; Founder and CSO, MTL, LLCMajor bacterial protein production systems use plasmids as vectors. We present new data demonstrating high levels of proteins expression without plasmids. We show how this leads to faster development times, and can substantially lower cost of production.







JANUARY 19-20





























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TUESDAY, JANUARY 20


ENGINEERING GENES AND ENHANCING EXPRESSION SYSTEMS

8:30 Chairperson’s RemarksJoseph Shiloach, Ph.D., Head, Biotechnology Core Laboratory, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health

8:35 Gene and Protein Sequence Optimization for High-Level Production of Fully Active and A-Glycosylated Lysostaphin in Pichia pastorisKarl E. Griswold, Ph.D., Associate Professor, Bioengineering, Thayer School of Engineering, Dartmouth CollegeIn the ongoing battle against drug-resistant bacterial pathogens, there is growing interest in lytic cell wall hydrolases as prospective therapeutic agents. Facile and scalable production of such bactericidal enzymes can, however, bottleneck both early stage discovery and downstream development of promising therapeutic candidates. Here, we describe an efficient, high-yield Pichia pastoris expression system for lysostaphin, a potent antibacterial agent for Staphylococcus aureus, including MRSA and other resistant strains.

9:05 Enhance CHO Productivity Using RNAi TechnologyJianxin Ye, Ph.D., Principal Scientist, Merck & Co., Inc.A siRNA library was screened to identify siRNA, which improves monoclonal antibody productivity from established CHO cell lines. The lead siRNA was then incorporated in the expression platform to enhance protein expression. The mechanism of some lead siRNA was also explored and one of the siRNA increases the mRNA level.

9:35 New Tools for Protein Solubility Sponsored by

David Mead, Ph.D., Founder & CSO, Lucigen CorpA panel of 24 solubility and expression enhancing fusion partners has been developed to simultaneously test multiple tags within the context of a single promoter, vector and host system. In addition, a novel yellow fluorescent protein significantly enhances solubility and expression while providing an instant visual report of the amount of soluble, active protein. This system permits rapid, simultaneous screening of multiple factors demonstrated to improve solubility and/or expression in a high throughput format using a robust enzyme-free cloning platform. The utility of the panel was proven in expressing soluble, active LRRK2, a very challenging biomarker for Parkinson’s disease.


11:00 Production of Recombinant Protein by a Novel Oxygen-Induced System in E. coliJoseph Shiloach, Ph.D., Head, Biotechnology Core Laboratory, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of HealthThe SoxRS regulon of E. coli is activated in response to elevated dissolved oxygen concentration likely to protect the bacteria from oxygen damage. This activation might make this regulon a candidate for recombinant protein expression. To test this, the soxS promoter was cloned into the commercial pGFPmut3.1 plasmid and the expression of GFP was evaluated by increasing the dissolved oxygen concentration. After high-density growth, it was possible to express ~0.5g GFP/L representing 3.4% of the total bacterial proteins.

11:30 Expression of Soluble and Active Interferon Consensus in SUMO Fusion Expression System in E. coliJi-won Choi, Ph.D., Director, Scientific Evaluation, PolyTherics Ltd.We describe the optimisation of the production of IFN-con in E. coli as a soluble IFN-con. The recoded IFN-con gene was cloned into a SUMO expression vector downstream of the SUMO fusion protein under T7lac promoter and expressed in E.coli SHuffle® strain. The fusion protein was efficiently expressed in soluble form and IFN-con remained stable following removal of the SUMO fusion partner. Also, antiviral activity of the produced IFN-con proved greater than IFN α-2a activity.

12:00 pm AutoPlasmid MEA for Automated Mini, Midi and Maxi DNA Preps

Sponsored by

Chris Suh, Ph.D., Business Development Manager, PhyNexus, Inc.Numerous processes in pharmaceutical development require the use of higher throughput plasmid DNA purification. The majority of issues encountered in Mini, Midi, and Maxiprep purification kits involve flocculate removal, and there is currently no easy way to produce large amounts of plasmid DNA without complicated and time consuming clarification steps. The AutoPlasmid MEA instrument provides a fully automated solution in plasmid purifications, allowing Mini, Midi, and Maxiprep plasmid purifications to be performed on a single instrument.

12:30 Session Break







JANUARY 19-20





























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2:00 BuzZ Session A









17th Annual Recombinant Protein Expression and Production

Achieving Quality and Quantity

JANUARY 21-22





























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Biopharmaceuticals currently represent the fastest-growing sector of the pharmaceutical industry, driven by a rapid expansion in the manufacture of recombinant protein-based drugs. To meet the demand, it is crucial to increase the throughput of expression, production and purification processes and systems. The Recombinant Protein Expression and Production conference explores the newest data and innovations relating to the best choices in hosts/systems, as well as ways to revive existing systems and make them work more effectively to produce the quality and quantity of the desired biotherapeutic.

TUESDAY, JANUARY 20


2:00 BuzZ Session A







ENHANCING EXPRESSION AND PRODUCTION: PROCESS DEVELOPMENT

8:15 Chairperson’s Opening RemarksStephen R. Hughes, Ph.D., Research Molecular Biologist, Renewable Product Technology Research Unit, U.S. Department of Agriculture, Agricultural Research Service, National Center for Agricultural Utilization Research


8:20 In situ Protein Expression for High-Throughput Protein Microarray StudiesJoshua LaBaer, M.D., Ph.D., Director, Virginia G. Piper Center forPersonalized Diagnostics and Virginia G. Piper Chair, Personalized Medicine, Biodesign Institute, Arizona State UniversitySelf-assembling protein microarrays can be used to study protein-protein interactions and protein-drug interactions, to search for enzyme substrates and as tools to search for disease biomarkers. In particular, recent experiments have focused on using these protein microarrays to search for autoantibody responses in cancer patients. These experiments show promise in finding antibody responses that appear in only cancer patients. New methods using click chemistry-based reagents also allow the application of these arrays for discovering new substrates of post-translational modification.

9:00 Optimizing the Cell Line Development Workflow to Support Accelerated TimelinesChristine DeMaria, Ph.D., Associate Director, Therapeutic Protein Expression, Global Biotherapeutics Development, Sanofi USTo support program timelines, we have further optimized our workflow to generate high-titer CHO cell lines. This included modifying the process for selection and sorting of transfected pools and more stringent 96-well plate based screens. These changes resulted in higher-titer clones being identified sooner, supporting more rapid development and quality assessment of lead clones. Workflow optimization case studies will be presented.

9:30 Hijacking E. coli’s Heat-Shock Response Boosts Protein ProductionXin Zhang, Ph.D., Burroughs Wellcome–CASI Fellow, Scripps Research Institute The production of high-yield and high-quality recombinant proteins from Escherichia coli is highly desirable for both academic and industrial settings. In this talk, I will describe a generally applicable method for this purpose, using a transcriptionally reprogrammed E. coli by over expressing a heat-shock response transcription factor. Similar strategies of hijacking stress-responsive pathways should be useful to enhance cellular protein folding capacity and improve recombinant protein production in other cell types.


10:50 Effect of Ambient Light on Monoclonal Antibody Product Quality during Small-Scale Mammalian Cell Culture Process in Clear Glass BioreactorsMary Mallaney, Engineer II, Purification Development, Genentech, Inc.During a monoclonal antibody (mAb) small-scale cell culture process, a larger-than-expected difference was observed in the charge variants profile of the harvested cell culture fluid between the 2L and larger scales. It was determined that ambient laboratory light induces photoreactions causing an increase in acidic variants. Our data clearly indicate that care should be taken when glass bioreactors are used in cell culture studies during mAb production.






JANUARY 21-22





























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11:20 Integrated Automation for Continuous High-Throughput Synthetic Chromosome Assembly and Transformation to Identify Improved Yeast Strains for Industrial Production of Peptide Sweetener BrazzeinStephen R. Hughes, Ph.D., Research Molecular Biologist, Renewable Product Technology Research Unit, U.S. Department of Agriculture, Agricultural Research Service, National Center for Agricultural Utilization ResearchWe describe a one-step construction of a synthetic yeast artificial chromosome (YAC) containing the optimized ORFs in a polyprotein cassette for expression of multiple genes such as those for enzymes in metabolic pathways or for valuable peptide or protein coproducts behind an optimized promoter with custom expression fusion tags selected for desired expression levels and protein locations inside or outside the industrial strain.

11:50 The Rapid Generation of Multiple Antibody Toxin Conjugates: Expression in Eukaryotic AlgaeMiller Tran, Ph.D., Senior Scientist, Lead Discovery, Verdant Therapeutics, Inc.Over the last decades, targeted antibody therapies, including antibody drug conjugates and recombinant immunotoxins, have garnered increasing attention. However, their production has become increasingly complicated and expensive. To overcome the challenges associated with the production of targeted therapies, eukaryotic algae are being used to produce recombinant immunotoxins that overcome the shortcomings of established technologies. With media cost in the cents per liter, the potential of algae is now being realized.

12:20 pm Taking Protein Production in Escherichia Sponsored by coli to the Next LevelDavid Vikström, Ph.D., CTO, Xbrane BioscienceThe bacterium Escherichia coli is a widely used host for the production of proteins. However, protein production in E. coli is usually a challenge. Therefore, Xbrane has developed new titratable expression systems. These systems allow optimizing the production of both routine and difficult targets in E. coli. The expression systems in combination with optimization and strain engineering are really taking the protein production in E. coli to the next level.

12:50 Session Break

1:00 Luncheon Presentation: Best of Both Worlds: Innovative Microbial System Leverages the Advantages of Both Bacterial and Mammalian Manufacturing

Sponsored by

Kristin DeFife, Ph.D., Director, Biologics Manufacturing, Ajinomoto Althea, Inc.Reach high expression using a microbial system and overcome challenges associated with E. coli including aggregation, lengthy purification and inefficient refolding processes. The Corynex® system secretes properly folded, biologically active proteins into the extracellular fermentation broth like mammalian cells which eliminates multiple purification steps, lowering cost and speeding time to market.

DIFFICULT EXPRESSION AND PRODUCTION: MEMBRANE PROTEINS

2:00 Chairperson’s RemarksIan Hunt, Ph.D., Group Leader, Proteomic Chemistry and Head, Protein Sciences, Novartis

2:05 Designer Surfactant-Like Self-Assembling Peptides for Membrane Protein Purification and StabilizationSotirios Koutsopoulos, Ph.D., Research Scientist, Center for Biomedical Engineering, Massachusetts Institute of TechnologyMembrane proteins are integral proteins of the cell membrane and are directly involved in the regulation of many biological functions and in drug targeting. However, our knowledge of membrane proteins is limited due to difficulties in producing sufficient quantities of soluble, functional and stable receptors. Designer, surfactant-like peptides may be used to extract the protein from the cell membrane and stabilize the protein outside the membrane bilayer for further studies.

2:35 Production of Human Integral Membrane Proteins in Mammalian CellsJames D. Love, Ph.D., Director, Technology Development, Biochemistry, Albert Einstein College of MedicineIntegral membrane proteins are key targets in the understanding of health and human disease, yet producing functional material in great enough quantities for structural studies remains a formidable task. This talk highlights the expression technologies under development that will greatly aid high-throughput efforts to produce functional human membrane proteins and complexes, specifically GPCRs, for a plethora of structural techniques.

3:05 Advances for Maximizing Protein Yields in HEK293 and CHO Transient Expression

Sponsored by

Henry C. Chiou, Ph.D., Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher ScientificIncreasing focus on complex proteins in advanced research elevates the need for higher yields from mammalian transient expression systems. To achieve this objective while maintaining speed, simplicity and ease-of-use requires coordinated development and synergy between novel high-capacity media, higher productivity HEK293 and CHO cells, and higher performance transfection reagents. This presentation will demonstrate the effectiveness of this “holistic” approach to achieve gram-level protein yields by transient expression.

3:35 The Need for Technology Platforms in USP Development

Sponsored by

Hugo de Wit, CEO, CellcaAre you faced with budget reductions for USP development or even anticipated to do “more with less”? Do you see increasing expectations from market and management regarding reducing timelines and cost of goods attributes? We demonstrate how a robust USP platform can help you meet these challenges.






JANUARY 21-22





























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3:50 Bypassing Inclusion Body Formation in E. coli: Maximizing Soluble Expression of Complex Multimeric Proteins

Sponsored by

Mark Valasek, M.D., Ph.D., Co-Founder and Scientific Director, AbSciCytoplasmic expression of large complex proteins in E. coli is limited primarily due to protein aggregation and the corresponding refolding process. These costly aggregates form when proteins are expressed too quickly and strongly. AbSci has developed a tightly regulated, dual titratable expression system that is able to homogenously induce high levels of complex multimeric proteins in a soluble and active form.






ENGINEERING EXPRESSION AND PRODUCTION: CHO CELLS

8:30 Chairperson’s RemarksHelene Faustrup Kildegaard, Ph.D., Co-Principal Investigator, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark

8:35 Omics-Guided Cell Line Engineering: Reducing High Mannose by Overexpressing N-Glycosylation Pathway RegulatorsShivani Gupta, Associate Scientist, Cell Line Development, Amgen, Inc.High mannose is a critical quality attribute for recombinant therapeutic monoclonal antibodies that can impact biological activity by influencing Fc-mediated effector functions, product stability, clearance rate and safety. In this study, we identified key genes whose expression is correlated with lower levels of high-mannose glycans in antibody-producing CHO cell lines.

9:05 Improving CHO Cell Factories Using Systems Biology and Genome Editing TechnologiesHelene Faustrup Kildegaard, Ph.D., Co-Principal Investigator, Novo Nordisk Foundation Center for Biosustainability, Technical University of DenmarkNew opportunities are arising to improve CHO cell factories using emerging systems, biology data and efficient genome editing technologies. Here, our recent efforts in improving CHO cell factories by genetic engineering using mainly genomics and transcriptomics data combined with CRISPR Cas9 technology will be presented.

9:35 ESETEC® 2.0: New Generation of E. coli Secretion Technology for the High-Yield Production of Fabs Sponsored by

Andreas Anton, Ph.D., Head, Bioprocess Development, Wacker Biotech GmbHWACKER has profoundly refined its patented ESETEC® E. coli based system for the manufacture of biopharmaceuticals. Targeted genetic modifications and process optimization measures led to the development of new, extremely productive cell lines and fermentation procedures. ESETEC® 2.0 is now able to produce several grams per liter of secreted Fabs.


10:50 Identifying Targets for Engineering Protein Secretion in CHO Cells with Systems BiologyNathan E. Lewis, Ph.D., Assistant Professor, Lab of Systems Biochemistry and Cell Engineering, University of California, San DiegoOur recent whole-genome sequencing efforts for CHO have enabled the construction of systems biology models. We now use such models for detailed analysis of -omics data from CHO to deepen our understanding of differences between host cell lines and to develop predictions for enhancing cell growth, protein modification and protein secretion.

11:20 Overexpression of MicroRNAs Enhances Recombinant Protein Production in CHO CellsWan Ping Loh, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR)Cell engineering using microRNAs (miRNAs) is an emerging strategy to increase recombinant protein production in mammalian cells. We identified miRNAs that were differentially expressed between high- and low-mAb-producing CHO cell clones. Stable overexpression of these miRNAs in a CHO-mAb cell line resulted in increased IgG production of ~30% in pools and up to 140% in clones, without major changes in product aggregation and N-glycosylation profile.

11:50 Use of CHO Cell Engineering for Improved Protein SecretionPierre-Alain Girod, Ph.D., CSO, SelexisTypically in mammalian cell-based production systems, there is an inverse relationship between cell proliferation rate and cell-specific recombinant protein production. Yet cellular resources can be redirected from cell biomass accumulation to recombinant protein synthesis by changing the physico-chemical environment of the cells. But folding and assembly are highly protein-specific. To increase the functional capability of the cells and thus volumetric product titer, we engineered the host cells by increasing the rate of protein folding and assembly.







JANUARY 21-22

12:30 Luncheon Presentation: Looking to Achieve Amazing TPP Yields in CHO and HEK293? Introducing FectoPRO™, a Novel Powerful Transfection Solution

Sponsored by

Mathieu Porte, MSc, Bioproduction Project Leader, R&D, Polyplus-transfection®

Low transfection efficiency of CHO cells is a major bottleneck hampering Transient Protein Production (TPP). Polyplus-transfection®, with its 10+ year expertise in transfection, has developed a novel technologically advanced transfection solution specifically designed for bioproduction. FectoPRO™ outperforms currently available PEI-based and lipid-based transfection reagents. We will present data and protocols leading to unmatched protein and antibody yields in CHO and HEK-293 cells.






























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2nd Annual Transient Protein Production

From Small-Scale to Large-Scale

JANUARY 22-23





























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Speed, limiting risk and protein quality are known advantages of transient protein production (TPP). Transient expression achieves fast process times and high yields because the gene of interest is simply transfected via a plasmid into the respective producer cells. Protein quality and posttranslational modification benefit from recombinant protein expression in mammalian producer cells (CHO and HEK293) or alternative hosts, but rocketing demand for recombinant proteins requires improved TPP technologies. The Transient Protein Production conference convenes protein expression specialists who share differences, tradeoffs, advantages and improvements in producing recombinant proteins in transient production systems.




Sponsored by


Low transfection efficiency of CHO cells is a major bottleneck hampering Transient Protein Production (TPP). Polyplus-transfection®, with its 10+ year expertise in transfection, has developed a novel technologically advanced transfection solution specifically designed for bioproduction. FectoPRO™ outperforms currently available PEI-based and lipid-based transfection reagents. We will present data and protocols leading to unmatched protein and antibody yields in CHO and HEK-293 cells.


CHO CELL LINES

2:00 Chairperson’s Opening RemarksDominic Esposito, Ph.D., Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc.


2:05 Application of UPR Response and Cell Cycle Checkpoint Engineering for CHO Cell Line ConstructionTakeshi Omasa, Ph.D., Professor, Institute of Technology and Science, University of TokushimaThe construction of high-producing CHO cells is an important issue for therapeutic protein production. We introduce our new approach for Unfolded Protein Response (UPR)-based cell line construction and accelerating transgene amplification system using cell cycle checkpoint engineering.

2:45 Protein Production by PEI-Mediated CHO TransfectionYves Durocher, Ph.D., Team Leader, Protein Production, Biologics and Subsequent Entry Biologics, Life Sciences - NRC Human Health Therapeutics Portfolio, National Research Council CanadaWe present data describing our CHO-EBNA1 transient expression platform for the production of various proteins. For more difficult-to-express proteins or proteins needed in large quantities, we also developed an inducible CHO pool platform that allows the generation of stable pools expressing high levels of monoclonal antibodies in less than 3 weeks post-transfection. This platform can also be used to derive stable CHO clones for manufacturing therapeutic r-proteins candidates.

3:15 Systems-Based Approaches for Enhancing Transient Protein Expression in CHO CellsJonathan Zmuda, Ph.D., Associate Director, R&D, Thermo Fisher ScientificCHO cells are known to express lower levels of transient proteins than HEK293, but are desirable for biopharmaceutical development since they are the most commonly used cell type for bioproduction. To address the need for a higher-expressing transient CHO platform, systems-based approaches were utilized whereby the latest advances in cell culture media and feeds, transfection reagents and enhancers were optimized through multifactorial DoE to generate a complete solution for enhanced protein expression with titers comparable to HEK293-based systems.

3:45 Rapid Protein Production with Flow Sponsored by Electroporation: Higher Titers, Cell Type Flexibility and ScalabilityJames Brady, Ph.D., MBA, Director, Technical Applications, MaxCyte, Inc.As protein therapeutics grow, future success will depend on the ability to be flexible regarding cell type and the ability to quickly scale up or scale down. In this presentation, data from MaxCyte flow electroporation will be presented demonstrating its scalability, high transfection efficiency and cell viability of commonly used cells, including CHO-S, HEK293 and insect cells, and production of antibody titers >1 g/L in 2 weeks.







JANUARY 22-23





























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5:00 High-Throughput mAb Expression and Protein A Purification Platform Based on Transient CHOYashas Rajendra, Ph.D., Research Scientist, Biotechnology Discovery Research, Eli Lilly and CompanyWe developed a PEI-mediated transient CHO-GS KO expression system that can generate mAb titers up to 0.5 g/L. We also developed a semi-automated Protein A purification process. Using a single liquid handling robot, up to 72 unique mAbs can be simultaneously purified from 24DWP (deep-well plate) transfections. This process yields 0.3 mg-1 mg of high-quality mAb at concentrations >0.5 mg/ml in a standard formulation buffer, enabling rapid characterization of mABs in multiple assay formats.

5:30 PANEL DISCUSSION: CHO Cells: Transient, Stable or BothRecombinant protein expression in mammalian producer cells such as CHO offers significant advantages in protein quality and post-translational modification. Expression can be transient, which achieves fast process times and high yields, or stable, which takes greater time to convert but offers other gains. This panel convenes experts to share insights into the benefits and tradeoffs of meeting the demand for recombinant proteins by engineering transient or stable cell lines or a combination of both.Moderator: Dominic Esposito, Ph.D., Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc.Panelists: James Brady, Ph.D., MBA, MaxCyte, Inc.Yves Durocher, Ph.D., National Research Council CanadaTakeshi Omasa, Ph.D., University of TokushimaYashas Rajendra, Ph.D., Eli Lilly and CompanyJonathan Zmuda, Ph.D., Thermo Fisher Scientific


FRIDAY, JANUARY 23


ENGINEERING HIGHER TRANSIENT EXPRESSION IN ALTERNATIVE HOSTS

8:30 Chairperson’s RemarksRichard Altman, MS, Research Scientist, Molecular Sciences, Alexion Pharmaceuticals

8:35 Enhancement of Transient Protein Expression via Modulation of Intracellular Biomarkers and Discovery of Novel Non-Viral Delivery VehiclesKaushal Rege, Ph.D, Associate Professor, Chemical Engineering and Faculty, Biomedical Engineering and Biological Design, Arizona State UniversityWe describe a dual strategy involving discovery of novel transfection reagents

and inhibition of key intracellular biomarkers for enhancing transient protein production in mammalian cells. Several candidates from novel polymer and lipopolymer libraries demonstrated higher transgene expression efficacies compared to polyethyleneimine. In addition, cell-based studies resulted in the identification of several small-molecule protein inhibitors that enhanced polymer-mediated transgene expression.

9:05 Selected Oral Poster Presentation: Rapid Plant-Made Pharmaceutical Platform for Anti-Viral Monoclonal AntibodiesSylvain Marcel, Ph.D., Senior Scientist, Molecular Biology, Caliber BiotherapeuticsCaliber Biotherapeutics, the world’s largest plant-made pharmaceutical facility, has successfully and rapidly manufactured three antiviral monoclonal antibodies sufficient for small animal studies in less than two months using its proprietary transient plant expression platform. An independent survival rate study performed in mice infected with dengue demonstrated excellent comparability between the plant-made anti-DENV and its mammalian-made counterpart. This study demonstrates rapid manufacturing of therapeutics at Caliber resulting in plant-made material that is as biologically potent as the CHO-made product.

9:05 Selected Oral Poster Presentation: Enhanced DNA Delivery and Subsequent Plasmid Maintenance in a CHOK1SV Expression System for Improved Transient Recombinant Protein YieldsJames Budge, Research Scientist, Centre for Molecular Processing, University of KentTwo potential bottlenecks in transient gene expression (TGE) derive from low DNA transfection efficiencies and the cell’s inability to maintain extra-chromosomal plasmid DNA. In this study a novel nuclear localisation signal (NLS) has been identified and utilised to improve transfection efficiency and, furthermore, the Epstein Barr Nuclear Antigen 1 (EBNA1) system has been employed and assessed in terms of plasmid maintenance. Finally, these approaches have been implemented simultaneously in an attempt to multiply product yield.

9:35 Baculovirus-Mediated Transduction of HEK-293 and CHO Cells for the Large-Scale Expression of Recombinant Immunoglobulin and Virus-Like ParticlesChristopher W. Kemp, Ph.D., President, Kempbio, Inc.Baculovirus constructs with mammalian promoters (BacMam) are efficient transduction agents for a variety of cell lines. We report on the development of a large-scale protein expression platform using serum-free cultures of HEK-293 and CHO-S. Process optimization has increased productivity and rIgG expression levels have reached 140 mg/L at a scale of 10-liters. Influenza VLPs have been successfully produced using a tripartite BacMam transduction in shake-flask culture and in stirred-tank bioreactors.







JANUARY 22-23

11:00 Glycoengineering in the Baculovirus-Insect Cell SystemDonald Jarvis, Ph.D., Professor, Molecular Biology, University of Wyoming; President, GlycoBac, LLCWe focus on our ongoing efforts to create new baculovirus-insect cell systems for improved recombinant glycoprotein production. We have knocked in higher eukaryotic genes and knocked out endogenous functions to obtain human-type glycosylation and eliminate immunogenic sugar epitopes.

11:30 Modifications to the Baculovirus/Insect Cell Expression System to Enhance Production of Prenylated ProteinsDominic Esposito, Ph.D., Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc.To accelerate structural studies of KRAS and attempts to identify potential therapeutics, we have developed a modified baculovirus platform to permit high-yield production of prenylated RAS proteins with high accuracy and quality. We discuss the enhancements to the system and how they can be applied to high-level production of other clinically relevant prenylated proteins.

12:00 pm High-Level Transient Expression of Protein Biologics in Plants through AgroinfiltrationQiang (Shawn) Chen, Ph.D., Associate Professor, The Biodesign Institute and School of Life Sciences, Arizona State UniversityThis talk focuses on the high-level expression of protein-based biologics in plants through transient expression. Current expression vectors and agroinfiltration strategies will be discussed with virus-like particle (VLP)-based vaccines and monoclonal antibody (MAb)-based therapeutics as examples.


1:00 Session Break


ESTABLISHING FLEXIBLE PROTEIN PRODUCTION

2:00 Chairperson’s RemarksHenry C. Chiou, Ph.D., Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific

2:05 The Speed and Flexibility of Scalable Transient Protein Production: Meeting the Demands for a Wide Variety of Early Discovery Studies and ApplicationsRichard Altman, MS, Research Scientist, Molecular Sciences, Alexion PharmaceuticalsThe establishment of a robust and reproducible transient protein production facility provides critical support to drug discovery efforts. The genesis of our scalable transient protein production efforts will be briefly reviewed. The direct impact to our overall drug discovery efforts will be discussed through case studies highlighting the variety of both applications (functional studies, reagent production, etc.) and scale (microtiter plate to WAVE bioreactor) that our system provides.

2:35 Nimble Protein Production from Mammalian Cells in a Mid-Size Research SettingThomas Cameron, Ph.D., Associate Director, Biologics Drug Discovery, Research Division, Biogen IdecIn the Research division of Biogen Idec, we express hundreds of recombinant proteins per year ranging from 1mg to 10g. To fit a variety of needs, various systems are used, including CHO and HEK 293, transient and stable pools. Key aspects of our flexibility and philosophy will be discussed with an emphasis on recent use of the GnTI-deficient HEK 293 cells.

3:05 Toolbox to Streamline the Identification of Antibody Drug CandidatesGiovanni Magistrelli, Engineer, Head, Protein Generation, NovimmuneMonoclonal antibody generation requires high-quality antigen and relevant functional assays. We combined several innovative steps for cloning, expression, purification and immobilization of active recombinant proteins. We used mammalian episomal systems and disposable bioreactors to produce in vivo biotinylated proteins facilitating phage display selection and assay development. We also designed novel vectors streamlining cloning and IgG purification. Such toolboxes supporting the discovery of monoclonal and bispecific antibodies will be presented.

3:35 Utilization of a HT Transient Expression Platform to Enable Rapid and Predictive Candidate SelectionJennitte Stevens, Ph.D., Principal Scientist and Group Leader, Mammalian Expression Group, Therapeutic Discovery - Biologic Optimization, Amgen, Inc.I describe how we utilize a high-throughput transient expression platform during the discovery phase of the drug development process to assess early manufacturability of candidates, using cellular attributes such as titer, cell growth, molecular profiling and high-content analysis of secretion bottlenecks. Case studies will be shown. This process allows us to rapidly gather data on large panels of candidates to select the best molecules to move to development.






























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PIPELINE 5: ANALYTICS & IMPURITIES

Inaugural Characterization of ADCs, Bispecifics and New Biotherapeutics

Improving Prediction, Screening and Characterization of New Biologics

JANUARY 19-20





























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With the wave of new biotherapeutics formats flooding the discovery and development pipelines, there is an increasing need for better and faster characterization tools and strategies, and improved biomolecular and biophysical assays for the new biotherapeutics. The new Characterization of ADCs, Bispecifics and New Biotherapeutics conference will present high level case studies and strategies on improving characterization of ADCs, bispecifics, and novel protein formats such as domain antibodies. Cutting-edge tools, research findings and unpublished data will be featured at this forum.

SUNDAY, JANUARY 18




MONDAY, JANUARY 19


DEVELOPING A SUCCESSFUL ANALYTICAL STRATEGY FOR CANDIDATE SELECTION AND

OPTIMIZATION

9:00 Chairperson’s Opening RemarksJohnson Varghese, Ph.D., Senior Director, Head of Analytical Development, Shire HGT


9:10 Regulatory Perspective on Challenges in ADC DevelopmentWen Jin Wu, M.D., Ph.D., Senior Investigator, Division of Monoclonal Antibodies, Office of Biotechnology Products, OPS-CDER-Food and Drug AdministrationThe unique properties of ADCs create technical challenges that require careful CMC considerations. With the increase in ADC IND submission and recent approval of ADC products (Adcetris and Kadcyla), FDA has gained the in-depth knowledge regarding ADC development programs. This presentation will discuss FDA experience with ADC development and regulatory challenges, with special focus on CMC. A strategy for the development of the next generation of ADCs will also be discussed.

9:50 High-Throughput Developability Screening Methods Targeting Antibody Self/Cross InteractionEric Krauland, Ph.D., Senior Director, Antibody Discovery and Optimization, Adimab, LLCDevelopability issues, such as aggregation, low solubility, high viscosity and poor pK can be tracked to antibody self or cross interaction. The talk will review high-throughput methods to screen hundreds to thousands of antibodies, such as IgGs and bispecifics, within a single day to be compatible with early stage antibody discovery. Selecting for developability properties along with target biology in the earliest discovery stages aims to improve the efficiency of the overall development process.

10:20 Coffee Break

10:45 Analytical Characterization of Biologics for Candidate Selection and Optimization: Strategies and Case StudiesGuodong Chen, Ph.D., Senior Principal Scientist, Bioanalytical and Discovery Analytical Sciences, Research and Development, Bristol-Myers Squibb Co.Since the introduction of the first recombinant DNA-derived protein insulin in the 1980s, antibody market has shown a steady growth. However, there are significant analytical challenges in the characterization of antibodies. This presentation will highlight current challenges and new enabling analytical techniques for the selection and optimization of antibody candidates.

11:15 Challenges and Strategies for Establishing an Analytical Control Strategy for Novel Complex Recombinant BiotherapeuticsJohnson Varghese, Ph.D., Senior Director, Head of Analytical Development, Shire HGTEstablishing an analytical control strategy for recombinant non-mAb therapeutics that exhibit complex sets of post-translational modifications and size distributions can be extremely challenging due to lack or little prior knowledge regarding the impact of these attributes on safety and efficacy. In this presentation, strategies for developing a control strategy based on criticality assessments of product attributes from early to late stage development for novel molecules will be described.

ANTIBODY-DRUG CONJUGATES (ADCs): CHARACTERIZATION, DEVELOPABILITY AND

CONTROL STRATEGY

11:45 Light-Induced Degradation of Antibodies and Antibody-Drug ConjugatesChristian Schöneich, Ph.D., Professor and Chair, Pharmaceutical Chemistry, University of KansasAntibody-drug conjugates (ADCs) provide a promising approach to deliver large payloads to specific cell populations. However, chemical and physical instability may arise from the exposure of ADCs to light, as several ADCs contain drug conjugates, which may act as photosensitizer, and/or drug conjugation may trigger photosensitivity in general. This talk will focus on light-induced photodegradation of ADC mimics, designed to evaluate the light-sensitivity and degradation mechanisms of ADCs.






JANUARY 19-20





























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12:15 pm Molar Mass, Size and Interactions: Light Scattering Tools for Essential Biophysical Characterization

Sponsored by

Daniel Some, Ph.D., Director, Marketing, Principal Scientist, Wyatt Technology CorporationBiophysical techniques based on static and dynamic light scattering address many of the key analytical challenges in biotherapeutic R&D, from early candidate selection through scale-up, formulation, characterization and comparability studies. This seminar will review light scattering technology and instrumentation, then present select examples illustrating how Wyatt’s light scattering solutions facilitate rapid and effective development of biologics including mAbs, ADCs, PEGylated and other proteins.

12:45 Session Break


ANTIBODY-DRUG CONJUGATES (ADCs): CHARACTERIZATION, DEVELOPABILITY AND

CONTROL STRATEGY CONTINUES...

2:00 Chairperson’s RemarksShrikant Deshpande, Ph.D., Senior Director, Protein Chemistry, Biologics Discovery California, Bristol-Myers Squibb Co.

2:05 Challenges in ADC CharacterizationShrikant Deshpande, Ph.D., Senior Director, Protein Chemistry, Biologics Discovery California, Bristol-Myers Squibb Co.ADC characterization is a critical component of ADC research and development. Antibody characteristics, the structure of payloads - linkers and conjugation technologies influence the characterization of ADCs. In addition to QC requirements, ADC characterization will help explain the observed biological results in preclinical studies and provide guidance for product release specifications. In this presentation, challenges in the ADC characterization and translation of characterization strategies into product release assays will be discussed.

2:35 Cutting Edge Vibrational Spectroscopy for Protein Therapeutics

Sponsored by

Rina K. Dukor, Ph.D., President , CEO, BioTools, Inc.The use of FT-IR spectroscopy as a probe of secondary structure is now widespread throughout the biopharmaceutical industry. More recently, ROA (Raman Optical Activity) has been shown to indicate differences when none are observed with any other spectroscopic technique. In this presentation, we will discuss advances in four forms of vibrational spectroscopy as applied to structural studies of proteins.

3:05 Physical Characterization of ADC with Different LoadingsJianxin Guo, Principle Scientist, Biotherapeutics Pharmaceutical Sciences, Pfizer, Inc.Conjugation of an antibody to a drug can produce heterogeneous species that may have different physical stabilities and safety profiles. This talk will illustrate the effect of loading profiles on the physical characteristics and aggregation propensity of an ADC conjugated by thiol-maleimide chemistry. The application of biophysical techniques on the evaluation of hydrophobicity and structural stability will be elaborated.

3:35 Q&A with Session Speakers


4:15 Addressing Product Heterogeneity Challenges in ADC Biotherapeutics: Case StudiesNomalie Jaya, Ph.D., Senior Scientist, Seattle Genetics, Inc.ADCs produced through the chemical linkage of a potent cytotoxic agent (drug) to a monoclonal antibody (mAb) are likely to have product heterogeneities associated with drug- linker attachment site and conjugation process. ADC heterogeneity can be characterized and controlled through identification of critical quality attributes (CQAs), implementation of appropriate process controls and analytical tools. Here we discuss two case studies highlighting the implications of heterogeneities in an ADC analytical profile used for product quality monitoring; 1) Drug-linker charge and distribution on charge based purity and 2) ADC modality on size heterogeneity as measured under native and denaturing conditions.

4:45 Characterization of Antibody-Drug Conjugates and Control Strategies for Small Molecule ImpuritiesJuma Bridgewater, Ph.D., Scientist III, Analytical and Pharmaceutical Sciences, ImmunoGen, Inc.The challenge of characterizing ADCs is compounded by small molecule impurities present with the cytotoxic agent. Different control strategies are used for impurities based on their potential pharmacological effect, degree of protein modification and likelihood of clearance during processing. This talk describes different impurity control strategies and the characterization and release test methods that support them.






JANUARY 19-20





























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5:15 Biological and Analytical Characterization of ADCs Based on RNA Polymerase II Inhibiting ToxinsAndreas Pahl, Ph.D., CSO, Heidelberg PharmaToxic warheads of today’s ADCs are exclusively based on compounds acting on microtubules or DNA and seem to suffer from limitations in certain cancer indications and tumor cells. This talk will discuss a new generation of ADCs based on the toxin amanitin, a highly effective inhibitor of the eukaryotic RNA Polymerase II. This presentation will summarize the current status of this new payload and present a showcase about the biological and analytical characterization of amanitin based ADCs.


TUESDAY, JANUARY 20


ANALYTICAL CHALLENGES FOR BISPECIFICS AND NEW PROTEIN THERAPEUTICS

8:30 Chairperson’s RemarksVishal C. Nashine, Ph.D., Senior Research Investigator, Drug Product Science & Technology, Bristol-Myers Squibb Co.

8:35 Analytical Challenges of Next-Generation BiotherapeuticsHubert Kettenberger, Ph.D., Principal Scientist, Large Molecule Research, Roche Innovation Center PenzbergBispecific antibodies and antibody-cytokine fusion proteins can be expressed in single CHO cell lines and manufactured in clinical-grade quality despite their architecture which is more complex than standard IgGs. Format-specific side products may occur and need to be addressed during cell culture, purification, formulation and stability testing. Designated analytical assays accompany the whole process and ensure clinical-grade product quality.

9:05 Case Study of Grid Selection to Identify Bispecific Antibody Lead MoleculesHaiyan Jiang, Ph.D., Principal Scientist, Biotechnology Center of Excellence, Janssen R&DBispecific antibodies (bsAb) can act on dual targets for therapeutic applications. A grid of bsAb was generated using the controlled Fab-arm exchange method combining two panels of parental monoclonal antibodies. Strategies to prepare this grid as well as analytical methods including SE-HPLC, cation-exchange chromatography, hydrophobic interaction chromatography and capillary isoelectric focusing were established to identify the purity and charge-based characteristics of the bsAbs and parental homodimers in the grid.

9:35 Q&A with Session Speakers


11:00 Analytical Characterization of Protein-Peptide Conjugates to Support Vaccine Candidate Selection and Optimization: Challenges and Case StudiesQiong (Joan) Guo, Ph.D., Senior Principal Scientist & Analytical Group Leader, Vaccine Immunotherapeutics Research Unit, Pfizer, Inc.High sensitive and high resolution SEC-HPLC and mass spectrometric methods were developed to determine aggregate levels and peptide density of protein-peptide conjugates, respectively. Several case studies will be shown to demonstrate applications of both methods and challenges to support conjugation process optimization as well as formulation buffer screen to find desired buffers to stabilize conjugates. Effects of aggregates and peptide density on vaccine efficiency will be also shown.

11:30 An Experimental Design Approach to Effectively Address Formulation Challenges of a Fusion ProteinVishal C. Nashine, Ph.D., Senior Research Investigator, Drug Product Science & Technology, Bristol-Myers Squibb Co.Typically, during drug product development the target product profile (TPP) is not well-defined until later stages. Therefore, flexibility around the formulation composition is desirable in that it affords the opportunity to respond to an evolving target. Here we describe application of modern experimental design tools in rapid screening and optimization of formulation for a fusion protein to enable a clear risk-based decision making on drug product configuration.

12:00 pm Enjoy Lunch on your Own

2:00 BuzZ Session A









Inaugural Detection and Characterization of Particulates and Impurities

Rapid Tools and Strategies for Risk Assessment, Prediction and Characterization of Particles and Impurities from Products, Excipients and Processes

JANUARY 21-22





























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Particles and impurities can come from the products, any stage of processing or the packaging containers. The presence of particulates and impurities in the drug product can impact stability, safety, efficacy of the biomolecules and biologic products. Therefore, early understanding, detection and characterization of the impurities are critical to ensure safety and efficacy of the drug product for its intended duration of use. The Detection and Characterization of Particulates and Impurities conference provides a platform to explore novel tools and strategies to detect, characterize and carry out risk assessment of particles and impurities.

TUESDAY, JANUARY 20


2:00 BuzZ Session A







STRATEGIES FOR RISK ASSESSMENT AND CHARACTERIZATION OF IMPURITIES

8:15 Chairperson’s Opening RemarksKevin Mattison Ph.D., Principal Scientist, Bioscience Development Initiative, Malvern Instruments


8:20 Strategies for Characterization and Risk Assessment of Particles, Degradants and ImpuritiesSatish Singh, Ph.D., Research Fellow and Group Leader, Pfizer; Member, 787 Expert Panel, U.S. Pharmacopeial Convention (USP)Considerable attention is paid to the development of appropriate process and product controls and specifications for biotherapeutics. However, the knowledge of the impact of these parameters on the efficacy, PK/PD, safety, immunogenicity may not always be available. The presentation will examine use of strategies to incorporate risk assessment tools to evaluate criticality and drive characterization efforts.

ORTHOGONAL TOOLBOX FOR DETECTION AND CHARACTERIZATION OF SUBVISIBLE PARTICLES

9:00 Advances in the Detection and Characterization of Subvisible and Visible Particles and Other AggregatesAndrea Hawe, Ph.D., CSO, Coriolis PharmaThe talk gives an overview on current trends for aggregate, subvisible particle and visible particle characterization of biologics. Analytical methods that are routinely used for the quantification and partly also the identification of aggregates and particulates, such as dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), flow field flow fractionation techniques (AF4/HF5), analytical ultracentrifugation (AUC), resonant mass measurements (RMM), flow imaging microscopy, light obscuration and semi-automated visual inspection are presented.

9:30 Stability Challenges for Protein Therapeutics: Characterizing Aggregation Propensity and Further Formation of Subvisible ParticlesJoël Richard, Ph.D., Vice President, Peptides, CMC & Engineering, IpsenThe talk focuses on the appropriate biophysical methods to characterize protein aggregation propensity, show structural modifications of proteins, study the effect of excipients on these modifications and the subsequent mechanism of formation and characterization of subvisible particles. These techniques appear as powerful orthogonal tools to study protein structure alteration and aggregation, which are among the most striking issues, also potentially triggering immunogenic reactions. Clinical impact will also be discussed, as a potential safety issue.


10:50 Factors Influencing the Sizing and Counting of Subvisible Particles by Orthogonal MethodsRichard Cavicchi, Ph.D., Physicist, Bioprocess Measurements Group, National Institute of Standards and TechnologyWe have used a microfluidic device designed to perform simultaneous flow imaging (FI) and electrical sensing zone (ESZ) measurements to investigate causes for variation in the results from these subvisible particle characterization methods. We compare highly monodispersed, photolithographically produced rods and disks as well as protein particles to illustrate the large influence of shape and porosity. Results are compared to measurements on commercial instruments based on FI, ESZ, and light obscuration.






JANUARY 21-22





























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11:20 Identification and Characterization of Particles in Biopharmaceutical FormulationsMiguel Saggu, Ph.D., Associate Scientist, Late Stage Pharmaceutical Development, Genentech, Inc.Subvisible particles in biopharmaceutical formulations may impact potency and immunogenicity of biotherapeutics. Detection, quantification and identification of particles in high concentration formulations represents a challenge to date, in particular in the size range between 0.1 – 10 µm. Case studies to overcome these technical difficulties are presented here.

11:50 Polysorbate Degradation and Particles in Biopharmaceutical Formulations: Analytical Method to Characterize Particulates and DegradantsAnthony Tomlinson, Research Associate, Late Stage Pharmaceutical Development, Genentech, Inc.Polysorbates are commonly used surfactants in the formulation of biopharmaceuticals. It has been observed that upon long-term storage of formulated products, these surfactants can degrade into insoluble products (including free fatty acids) which can lead to increased subvisible and visible particle counts. A method for detection and quantification of total free fatty acids in placebo and active formulations will be presented here along with related case studies. Furthermore the utility of the method for identification and characterization of insoluble particle composition is discussed.


12:50 Session Break


RAPID SCREENING OF IMPURITIES FROM PRODUCT AND EXCIPIENTS

2:00 Chairperson’s RemarksAjit S. Narang, Ph.D., Principal Scientist, Drug Product Science & Technology, Bristol-Myers Squibb Co

2:05 Mechanism of Protein Aggregation and Sustained Effect of Shear During Drug Substance Manufacture on Drug Product StabilityAjit S. Narang, Ph.D., Principal Scientist, Drug Product Science & Technology, Bristol-Myers Squibb Co.Emerging data suggests that aggregation instability of a ready-to-use protein solution drug product may be linked to the shear forces experienced by the protein during downstream purification. Sustained conformational changes due to shear during processing may contribute to long-range hydrophobic protein-protein interactions in protein solutions. These interactions manifest in changes in the physicochemical properties of proteins including viscosity and

hydrodynamic diameter, in addition to protein aggregation during accelerated stability testing. These observations provide an experimental framework to develop an early predictor of aggregation instability.

2:35 Understanding Protein-Surfactant Interactions at Air-Water and Water-Solid InterfacesAlfredo R. Narvaez, Ph.D., Manager, Diluent Research & Formulation Group, Global Process Design R&D, Abbott, Inc.Modulation of protein presence at surfaces is of paramount importance for performance of reagents used in the clinical laboratory. In this work, protein displacement by surfactants was characterized at the air-water interface via oscillatory and dilational rheology and at the water-solid interface by the quartz crystal microbalance method. Discriminatory interactions between proteins and surfactants and the importance of surface properties are discussed.

3:05 Detecting Irreversible Aggregate Formation of an Antibody during Downstream ProcessingJoy Chen, Ph.D., Scientist, Process Development Analytical, OncoMed Pharmaceuticals, Inc.Analyzing in-process samples is important for process development of antibodies in order to achieve higher yield and acceptable product quality. By a variety of analytical methods including SEC and ion exchange chromatography, we observed standard low pH elution buffers of protein A chromatography can cause irreversible aggregate formation which appeared to be induced by antibody denaturation. This talk describes characterization efforts that facilitated changes in downstream processing to decrease aggregate formation and achieve higher product yield.

3:35 A High-Throughput Coupled Methodology for Quantitation of Iso-Aspartate Formation in Proteins and PeptidesAjit S. Narang, Ph.D., Principal Scientist, Drug Product Science & Technology, Bristol-Myers Squibb Co.Formation of isoaspartate arising from a spontaneous asparagine deamidation or aspartate isomerization is one of the most common non-enzymatic pathways of chemical degradation of protein pharmaceuticals. This presentation will discuss degradation mechanism and formulation strategies to mitigate isoAsp formation. Also, the utility of a fluorescence-based high throughput assay utilizing protein isoaspartyl methyltransferase assay to quantitate isoAsp in four protein/peptide formulations containing active species of varying scaffolds will be presented.









JANUARY 21-22





























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HIGH-THROUGHPUT DETECTION AND CHARACTERIZATION OF HCPs AND PROCESS-

RELATED IMPURITIES

8:30 Chairperson’s RemarksZ. Mark Plavsic, DVM, MVSc, Ph.D., DACVM, Chief Virologist & Head, Global Product Biosafety, Genzyme, a Sanofi Company

»FEATURED PRESENTATION8:35 Developing an Effective Viral Risk Mitigation Strategy for Bio-TherapeuticsZ. Mark Plavsic, DVM, MVSc, Ph.D., DACVM, Chief Virologist & Head, Global Product Biosafety, Genzyme, a Sanofi CompanyRecent discoveries of virus contamination of biological materials used to manufacture medicinal products and the challenges associated with the detection of adventitious agents highlighted a need for development of a comprehensive viral risk mitigation program. This talk will outline several key areas that should be considered in developing an effective viral safety program.

9:05 High-Throughput Analytical Development to Support BioProcess Monitoring: Case Study on SEC, Peptide Mapping and Host Cell Protein (HCP) AssaysJianxin Ye, Ph.D., Principal Scientist, Merck & Co., Inc.This work is focused on converting and optimizing traditional chromatography- and immunology-based analytical methods for biological drug compound analysis into high throughput assays using automation systems. A 5-min platform UPSEC assay, the hands-off TECAN platform for peptide mapping sample preparation, and a fully automated HCP assay on Gyrolab will be discussed. The high-throughput assay has significantly facilitated the throughput and improved assay performance in sensitivity, accuracy, precision and reproducibility.

9:35 Fast and Easy Generic Anti-CHO HCP Analysis, 96-Samples Assay-To-Data in 65 Minutes

Sponsored by

Rashi Takkar, MSc, Product Manager, Marketing, Pall ForteBio LLCPall ForteBio has teamed up with Cygnus Technologies to jointly develop an Anti-CHO HCP detection kit. While ForteBio Octet systems are the industry standard in easy and rapid high throughput analysis, Cygnus HCP ELISA kits are known for their broad HCP recognition and sensitivity. The new ForteBio-Cygnus Anti-CHO HCP kit will embody the best of both worlds. Users will achieve unparalleled time-to-results, streamlined and automated* workflow, enhanced dynamic range, and excellent precision and assay robustness. *Complete hands-off automated workflow achieved with the Octet HTX system.


10:50 Using Mass Spectrometry to Lift the Rug on Host Cell Proteins in Purified BiologicsVeronika Reisinger, Ph.D., Scientist, Analytical Characterization, Sandoz GmbHHCPs (host cell proteins) are commonly present in very low levels even in purified biologics. Their identification and quantification is useful for a better understanding of potential safety risks and to support ever more sophisticated purification strategies in the development of biotherapeutics. We present the development and application of highly sensitive and reliable mass spectrometry-based approaches to identify and quantify trace amounts of HCPs directly in samples containing purified biologics.

11:20 Proteomic Characterization of CHO Host Cell Proteins in Purification of Monoclonal AntibodiesAbraham M. Lenhoff, Ph.D., Professor and Chair, Department of Chemical and Biomolecular Engineering, University of DelawareThe presence of host cell proteins (HCP) in process streams during protein purification is routinely monitored by methods such as ELISA that provide a global measure of impurities present. However, knowledge of specific impurities can reveal anomalies such as the disproportionate presence of one or more individual impurities. This presentation will explore methods for characterizing HCP and their persistence in downstream processing, including by co-elution with product or by product association.

11:50 Development of Innovative High Throughput Analytics Technologies to Support Rapid Process DevelopmentJason Kuo, Scientist, Process Science, Boehringer Ingelheim Pharmaceuticals, Inc.With the ever increasing pressure to move drugs into the clinic faster, an emphasis has been placed upon a more streamlined higher-throughput process development especially in the areas of upstream cell culture and downstream purification. In order to support this increased turnover of molecules, the development of high throughput, fast turnaround analytics must also keep pace. This talk will discuss some of the current innovative technologies being implemented to address these needs.







3rd Annual Extractables and Leachables

Protecting Quality of Biologics by Ensuring Safety and Compatibility

JANUARY 22-23





























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In biopharmaceutical development and manufacturing, containers and even disposable equipment may leach chemicals into the product that can pose significant risks to product quality and potentially compromise the safety and efficacy of the biotherapeutics. The Third Annual Extractables and Leachables (E&L) conference brings together industry experts and thought leaders to share their insights on latest trends and guidelines, how to design analytical testing strategies for E&L, case studies on identification and risk assessment E&L in drug products and best practices for identifying potential leachables and their impact on biologics.





E&L GUIDANCE AND UPDATES FROM INDUSTRY AND GOVERNMENT WORKING GROUPS

2:00 Chairperson’s Opening RemarksDiane Paskiet, Ph.D., Director, Scientific Affairs, West Pharmaceutical


2:05 The Product Quality Research Institute (PQRI) and the Issue of Leachables and ExtractablesDaniel L. Norwood, M.S.P.H., Ph.D., Distinguished Research Fellow, Analytical Development, Boehringer Ingelheim Pharmaceuticals, Inc.The PQRI was created to facilitate interactions between regulatory authorities, academia and the pharmaceutical industry. In 2006, a PQRI working group published a recommendation document regarding leachables and extractables in Orally Inhaled and Nasal Drug Products which included safety thresholds for leachables. A second PQRI working group is creating a similar document for parenteral and ophthalmic drug products. This presentation summarizes the history and progress of PQRI related to leachables and extractables.

2:45 Determining Suitability of Plastic Manufacturing Systems for Therapeutic Products: USP Standard DevelopmentDesmond G. Hunt, Ph.D., Senior Scientific Liaison, U.S. Pharmacopeial (USP) ConventionTherapeutic products come into direct contact with plastic, glass and metal materials as the product is manufactured, stored and administered. Such contact may result in an interaction that may affect suitability for use (including its safety and efficacy) and therapeutic product may be adversely impacted by the interaction. The presentation will discuss USP’s effort to develop a new standard for plastic single-use manufacturing systems chapters, related to plastic materials characterization and extractables and leachable testing.

3:15 Initiatives by BPSA, ASME-BPE, ASTM Int’l and USP to Develop Standards and Standardized Approaches for Testing of Extractables from Single-Use EquipmentJerold Martin, MSc, Chairman, Technology Committee, BPSA; Senior Vice President, Global Scientific Affairs, Pall Life SciencesDespite the multiple advantages of single-use systems, these polymeric assemblies also elicit concerns for their potential to contribute leachable compounds that could potentially impact production, quality or safety of finished drug products. This presentation will describe how suppliers are working with users in ongoing efforts to establish practical and scientifically-based standardized procedures for component extractables testing and data reporting among suppliers while meeting user needs and regulatory requirements.

3:45 “Organic Impurity Profiling” for Drug Sponsored by Products versus “Leachable Studies”: What Is There to Learn?Karen Pieters, Project Manager, Extractables/Leachables, Toxikon CorporationOrganic Impurities are critical quality attributes of drug substances and drug products because they have the potential to affect the safety and efficacy. The origin of the impurity may also determine which guidelines (and associated control limits) to follow in the final evaluation of the drug impurity. The scientific results of Leachable Studies, and in particular “screening” leachable studies, may assist in establishing a broad organic impurity profile for drug products, at low concentration levels.


RISK ASSESSMENT OF TOXICOLOGICAL AND BIOCHEMICAL CHALLENGES

5:00 A Risk-Based Approach to Assess the Potential Impact of Extractables and Leachables Impurities from Single-Use Systems on the Safety and Quality of Biotechnology ProductsKim Li, Ph.D., DABT, MPH, Senior Manager, Product Stewardship Toxicology, Environment, Health, Safety & Sustainability, Amgen, Inc.Single-use systems (SUS) offer cost-effective new technologies for the manufacturing of biotechnology products. However, the extractables and leachables (E&L) impurities from the complex polymeric materials may pose risk to product quality and safety. The qualification of SUS presents challenges for regulators, pharmaceutical and device industries, as well as suppliers. This presentation proposes a risk-based strategy that integrates the understanding of materials science, process development, trace chemical analysis and toxicology assessments.






JANUARY 22-23





























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5:30 Biopharmaceutical Quality: Factoring Extractables into the Risk EquationDiane Paskiet, Ph.D., Director of Scientific Affairs, West PharmaceuticalCo-Developed by: Thomas Egert, Research Scientist, Boehringer Ingelheim GmbH & Co. KGMaterials that contact biologics during manufacturing, storage and those associated with the final packaging configurations can pose risks to quality. The risk of substances leaching into drug products should be realized. This case study will illustrate relationships between the chemistry of a rubber material and mass transport properties via modeling to enable identification and mitigation of hazards. This knowledge can feed into design of relevant studies to reduce the uncertainty of leachables and contribute to justifications for material selection.


FRIDAY, JANUARY 23


MATERIAL CHARACTERIZATION AND CONSIDERATIONS FOR A SUCCESSFUL

ANALYTICAL STRATEGY

8:30 Chairperson’s RemarksJerold Martin, MSc, Chairman, Technology Committee, BPSA; Senior Vice President, Global Scientific Affairs, Pall Life Sciences


8:35 The Use of Kinetic Migration Models to Facilitate Extractables and Leachables InvestigationsDennis Jenke, Ph.D., Baxter Distinguished Scientist, Technology Resources, Baxter Healthcare Corp.Leaching of organic substances from plastic materials by pharmaceutical solutions is kinetically constrained by migration and diffusion if equilibrium is not achieved during contact. Although these processes can be mathematically modeled, migration modeling is not widely accepted as a suitable means of establishing the suitability for use of plastic systems used in the pharmaceutical industry. This presentation contains examples of migration modeling in pharmaceutical applications and considers the use of modeling to optimize migration studies.

9:05 Incorporation of Extractables and Leachables into Design Controls for Drug/Device Combination ProductsCheryl LM Stults, Ph.D., Principal, C & M Technical Consulting, LLCLeachables and extractables have been important considerations for drug product container closure, packaging and delivery systems. Recent developments have placed an increased emphasis on compliance with design controls for drug/device combination products. This presentation will discuss the incorporation of extractables and leachables considerations into the relevant aspects of design controls for combination products.

9:35 Implementation of an Extractable and Leachable Program for Single Use Components and Systems Sponsored by

Chris Shields, Engineering & Validation Manager, Life Science, Saint-GobainThis presentation will highlight the current best practices for E&L testing for Single Use components and Systems. The varibility from analytical equipment (lab to lab and within lab), product (within lot and lot to lot)and preparation method (surface are, time, temperature, extraction fluids) will be presented and compared to extractable results from multi-site manufactured products and products from different manufacturers. A case study that compares optimized extractable testing results to industry recommendations for different product materials and form factors will be highlighted.


EXTRACTABLES AND LEACHABLES IN SINGLE-USE SYSTEMS

11:00 The Design and Data Interpretation of Extractable Studies of Polymeric Product Contact Materials in the Manufacturing of BiologicsPing Wang, Ph.D., MBA, Principal Scientist, Pharmaceutical & Material Sciences, DPD, PDMS, Janssen Research & DevelopmentWith the increased scrutiny from regulatory authorities, more attention has been paid to the design and data interpretation of controlled extractable studies of polymeric disposable materials. The key to the success is to ensure the study design and data interpretation is product and process specific. The lack of relevant E&L data from suppliers presents end-users a great challenge. Strategies of developing relevant extractable data and applying that in the toxicity evaluation will be discussed.

11:30 Managing the Risks of Leachables from Single-Use Processing Equipment- A Practical ApproachMichael Ruberto, Ph.D., Material Needs Consulting, LLCThis presentation will focus on the key questions to consider when selecting single-use equipment to ensure that it is safe and compatible with the drug. These questions take into account the chemical composition of the materials used to fabricate the components, the conditions of contact between the equipment and drug, as well as the solvating power of each ingredient used in the drug formulation.

12:00 BPOG Extractable Protocol- Case Studies with 6 Model SolventsKen Wong, Deputy Director, MTech/AP&T - Extractables & Leachables, Sanofi PasteurThe BPOG proposed standard extractable protocol has generated tremendous amount of discussion around the merits in terms of analytic and methodology. In this presentation, the results from recently concluded case studies with 2 components (bag and O-ring) using the proposed 6 model solvents will be presented. For the first time, this study will provide a clear value proposition of the protocol and comparison of the extractable profiles among 6 models solvents.






JANUARY 22-23





























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12:30 Important Considerations When Determining Filter Flush VolumesJennifer Juneau, Principal Scientist, BioTherapeutics Research and Development, Pfizer, Inc.Extractable and Leachable risk assessments are performed to evaluate all product contact components throughout the intended manufacturing process. Since the sterile filter is typically one of the last process components in contact with the product it poses a substantial risk. To minimize this risk and maintain the safety and quality of the product, filters are flushed to remove potential leachables and equilibrate the membrane preventing protein and surfactant adsorption effects.

1:00 Session Break


ENSURING STABILITY, SAFETY AND EFFICACY OF BIOPHARMACEUTICALS

2:00 Chairperson’s RemarksMarisa K. Joubert, Ph.D., Senior Scientist, Process & Product Development, Amgen, Inc.


2:05 Protein Aggregate Size: A Potentially Important Factor in Inducing an Immune Response in Both in vitro and in vivo Model SystemMarisa K. Joubert, Ph.D., Senior Scientist, Process & Product Development, Amgen, Inc.This presentation will discuss the impact of size of protein aggregates and will discuss in detail how it can play a potentially important role in inducing an immune response in both in vitro and in vivo model systems.

2:35 Examining the Leachable from Disposable Bioprocess Bag and Their Impact on Protein StabilityNina Xiao, Ph.D., Senior Research Associate, Late Stage Pharmaceutical Development, Genentech, Inc.Leachables from BioProcess Containers (BPCs) are source of process-related impurities that have the potential to alter product quality and affect patient health. In our studies, we observed IgG1 instability after storage in gamma irradiated single-use plastic BPCs. Analysis of protein’s quality attributes provided useful hints on potentially unidentified leachables induced by gamma irradiation of BPCs. Risk mitigation for preventing the effect of leachables on protein instability is discussed.

3:05 Surface Plasmon Resonance imaging (SPRi): A Multifaceted Tool for Protein Analysis in SerumMarinella Sandros, Ph.D., Assistant Professor, Department of Nanoscience, University of North Carolina at GreensboroSPRi is a label free surface - sensitive optical detection method for monitoring biomolecular interactions in real time with high throughput. Attendees will gain a better understanding of how SPRi can be used as a flexible tool to screen and study protein interactions and to assess stability in a complex environment. Since SPRi detection is a non-destructive method, retrieved proteins can be analyzed simultaneously with MALDI - TOF, fluorescence and electrochemistry.

3:35 Extractables/Leachables in Biopharmaceutical Manufacturing Processes: Sources, Risks, and Case StudiesKathryn McGohan, Associate Scientist II, Manufacturing Sciences & Technology, Bristol-Myers Squibb Co.Extractables/leachables pose many challenges and concerns in biopharmaceutical manufacturing processes. These challenges and concerns are significantly increased when single-use products are used. Successful management of extractables/leachables issues includes identifying the sources of the compounds, understanding the process requirements, and analyzing the potential risks. This talk will present several case studies where different strategies were used by cross-functional teams to identify and control the risks related to material extractables/leachables.




PIPELINE 6: PROCESS TECHNOLOGIES & PURIFICATION

2nd Annual Single-Use Technologies and Continuous Processing

Driving Value through Process Innovations

JANUARY 19-20





























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Single-use technologies and continuous processing are the two key drivers in the bioprocessing industry today. Although each can be implemented on its own, the combination of single-use with continuous processing promises to bring even greater flexibility and productivity gains. CHI’s Second Annual Single-Use Technologies and Continuous Processing conference highlights the risks, challenges, opportunities and strategies for implementing these two separate yet complementary technologies, and showcases how companies can drive value and increase productivity through these innovations.

SUNDAY, JANUARY 18




MONDAY, JANUARY 19


9:00 Chairperson’s Opening RemarksJ Christopher Love, Ph.D., Professor, Chemical Engineering, MIT


9:10 Driving Value through Manufacturing: Innovation in a Hybrid Biologics Manufacturing NetworkLawrence Weiner, Ph.D., Senior Director, Strategic Innovation, Biogen IdecHas the time come for disruptive innovation or will continued sustaining innovation be sufficient for the challenges of a changing industrial environment? To answer this question, we discuss Biogen Idec’s approach to a hybrid network of biologics manufacturing facilities. We also discuss a few opportunities for innovation within existing technologies and also share a few thoughts on continuous manufacturing as an alternate route to maximize productivity.

9:50 Towards Biomanufacturing on DemandJ Christopher Love, Ph.D., Professor, Chemical Engineering, MITThe delivery of biologic drugs to patients can be challenging in many regions of the world. Patients may live in remote regions, under-resourced areas, or face challenging circumstances such as natural disasters. The state-of-the-art approaches to manufacture biopharmaceuticals are not compatible with on-site, rapid manufacturing of treatments on demand. This talk will present Integrated and Scalable Cyto-Technology (InSCyT) as a platform for continuous, mobile production in a closed-system facility that includes PAT and QbD.

10:20 Coffee Break

RISK MITIGATION STRATEGIES FOR SINGLE-USE TECHNOLOGIES

10:45 A Lifecycle Approach to Management of Leachables & Extractables Related Risk in Processes with Significant Investment in Single Use ComponentsNaveen Pathak, MSc, Director, MS&T, Process Development & Manufacturing Science, ShireDetermination of cumulative risk under worst-case and likely-case scenarios will be demonstrated to be an important first step to establish the baseline safety profile of the process. A risk based approach to evaluate subsequent addition of new single use components will be discussed. The L&E data will be used to support creation of a library of well characterized single-use components that will support efficient new product introductions.

11:15 Extractables and Leachables from Single-Use Components - Can They Be Cleared through Ultrafiltration/Diafiltration (UF/DF) Process?Kate Lee, Ph.D., Process Development Engineer, Genentech, Inc.Studies were performed to quantify the clearance of common small molecules that are extracted or leached from single-use components in the unit operations upstream of the ultrafiltration/diafiltration (UF/DF) step by UF/DF process. Results from the study indicated clearance of defined extractbles/leachables in protein solutions. However, unexpected clearance phenomena were observed for specific groups of extractbles/leachables, which will be discussed.

11:45 Advances in Next Generation ManufacturingRobert Dream, PE, CPIP, CPMP, Ph.D., Principal, HDR Company Ltd.

12:15 pm Single-Use Bag Extractable Case Study: Sponsored by Lessons LearnedMike Johnson, Global Bioprocess Applications Manager, Entegris, Inc.A supplier’s ability to furnish extractable data to end users is of extreme importance. The protocols followed to produce such data are currently a point of discussion between suppliers and end users. In this case study, single-use bags were subjected to a portion of the BioPhorum Operations Group (BPOG) extractable protocol. This presentation documents the testing process, lessons learned, along with sample extractable data and trends from a supplier’s perspective.

12:45 Session Break






JANUARY 19-20





























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STANDARDS AND RECOMMENDATIONS FOR SINGLE-USE EQUIPMENT AND PROCESSES

2:00 Chairperson’s RemarksJames Dean Vogel, P.E., Founder & Director, The Bioprocess Institute; Chair, BPSA Particulates Workgroup

2:05 Implementation Strategies and Challenges for Single-Use at Clinical to Commercial Scale: Integrity Testing, Material Qualification, Handling RisksAdam Goldstein, MSc, Senior Manager, Technical Operations, Genentech, Inc.This talk will review areas that should be considered with respect to disposable implementation. Understanding industry standards, quality and manufacturing needs for use in commercial applications, standardization of designs, as well as handling risks at full scale will be discussed. This presentation will outline key issues and how industry associations, suppliers and end users are addressing some of these areas.

2:35 Driving Adoption of Single-Use into cGMP: Reflections of the BPOG End Users Disposable Working GroupDavid Pollard, Ph.D., Executive Director, Bioprocess Development, Merck Research Laboratories, Merck & Co., Inc.; Member, BioPhorum Operations Group (BPoG)Efforts to implement SUT into commercial operations often exposes gaps in expectations between Supplier and End User. Case studies will cover the expectations of key issues for materials, film properties, extractables, sterilization validation, assembly process, change notification and integrity testing. Examples will be presented for achieving a wide and consistent alignment on the support needed for GMP environments, providing the foundation for SUT standards , strengthen collaboration between end users and suppliers and improve SUT implementation.

3:05 BPSA Update:

Part 1: Recommendations for Testing, Evaluation and Control of Particulates from Single-Use Process EquipmentJames Dean Vogel, P.E., Founder & Director, The Bioprocess Institute and Chair, BPSA Particulates WorkgroupThis paper and documents serve two purposes: to serve as a guide to characterization and determination of levels and types of particles in SUTs as well as to recommend procedures to achieve minimal levels of particles in SUT.

Part 2: Industry Initiatives to Standardized Single Use Manufacturing – Extractables, Quality Agreements, System DesignsJerold Martin, MSc, Chairman, Technology Committee, BPSA, and SVP, Global Scientific Affairs, Pall Life SciencesThis presentation describes how suppliers and users are working together to establish standardized practices including component extractables testing and data reporting, quality agreements and system designs.



RISK-BENEFIT ANALYSIS AND PROCESSING CHALLENGES

4:15 Can Continuous Processing and Single-Use Benefit Vaccine Manufacturing?Ronald Neeleman, Ph.D., Senior Director, Vaccine Innovation and Virology Expert, Global Technology Innovation, Sanofi PasteurVaccine manufacturing has a long history and a great variety (attenuated, inactivated, subunits, conjugations, etc.) but the production methods have remained batch in stainless steel. This presentation will deal with the key drivers to move from batch to continuous processing in single-use systems. Based on examples and experience of Sanofi Pasteur, the technical, scientific, and business motivations will be addressed.

4:45 Continuous Upstream Processing – Single-Use Versus Standard TechnologiesBerthold Boedeker, Ph.D., Chief Scientist, GDD-Global Biologics-Biotech Development, Bayer Pharma AGContinuous processing for the production of biologics from mammalian cell culture is developing into a viable alternative to the standard fed-batch fermentation. One of the main advantages is to reduce the necessary equipment size so that production can be done in simple lab-like manufacturing infrastructure using disposable instead of hard-piped equipment. This talk will describe advantages and disadvantages of continuous upstream processing in general and will compare single-use with standard, non-single-use based set-ups.

5:15 Mitigating Scale-Up and Process Challenges for Microbial Fermentation in Single-Use Production VesselsRajesh Krishnan, Ph.D., Director, BioProcess Development, Gilead Sciences







JANUARY 19-20





























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TUESDAY, JANUARY 20


CONTINUOUS PROCESSING: NOVEL CONCEPTS AND IMPLEMENTATION CHALLENGES

8:30 Chairperson’s RemarksRonald Neeleman, Ph.D., Senior Director, Vaccine Innovation and Virology Expert, Global Technology Innovation, Sanofi Pasteur


8:35 mAbs for the Masses: Towards Automated Continuous Processing Enabled by Single-UseDavid Pollard, Ph.D., Executive Director, Bioprocess Development, Merck Research Laboratories, Merck & Co., Inc.Efforts towards the next-generation mAb processing will show the integration of new technology with single-use-enabled continuous processing. The combined efficiencies gained by continuous processing, using integrated upstream and purification, will be compared by economic criteria to current manufacturing methods. The impact of these novel approaches to the process fit towards flexible, multiproduct manufacturing facilities will be described.

9:05 Why Continuous Protein Production Is Protein FriendlyPeter Tiainen, Ph.D., Department Manager, Novo Nordisk Research Centre ChinaAn integrated protein production system where perfusion cultivation is combined with a continuous multi-step purification platform enables the passionate of proteins to establish close to perfect protein-friendly production processes. From the expressing cells to the capture column and then along to additional columns, the protein is transported without hold/storage tanks, circumventing questions of storage stability, etc. Additional reasons why continuous bioproduction should appeal to protein enthusiasts will also be shared.



11:00 Novel Concepts for Protein Production – Status and Path ForwardThomas Daszkowski, Ph.D., Vice President, Process Design & Optimization, Bayer Technology Services GmbHWe will present our in-house approach of a novel small-scale and highly flexible protein production process. The process itself can be operated in a hybrid or fully integrated continuous mode. In addition to the process, key elements in regard to the PAT and process control targets will be discussed.

The outlook will touch base on challenges which we need to overcome, such as ready-to-use process equipment, PAT and process control system as well as regulatory requirements.

11:30 Continuous High-Throughput Downstream Processing for BiologicsThomas Müller-Späth, Ph.D., Senior Scientist, Institute for Chemical and Bioengineering, ETH ZurichContinuous chromatography processes allow improved downstream process performance of biomolecules including higher throughput, improved stationary phase utilization, improved yield and reduced buffer consumption. Different process concepts, including SMB and MCSGP are reviewed in the context of integration into continuous downstream manufacturing and single-use devices. Application examples of mAb capture using a twin-column countercurrent capture SMB process and polishing using a twin-column ion-exchange MCSGP process are discussed in greater detail.

12:00 pm: Biovest — Single-Use Disposables Since the 80s

Sponsored by

Mark Hirschel, Ph.D., Senior Scientific Advisor, Biovest International, Inc.Biovest offers a proprietary line of hollow fiber bioreactors that produce milligram to kilogram quantities of secreted proteins, including monoclonal antibodies and recombinant proteins, viruses or VLPs, and can be used for whole cell expansion. Our technology achieves high cell densities, is highly automated and operates as a closed system.

12:30 Session Break


2:00 BuzZ Session A









7th Annual Protein Purification and Recovery

Streamlining Processes

JANUARY 21-22





























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The Protein Purification and Recovery conference explores how experts are innovating processes to achieve purity, and provides an overview of the current tactics and technologies for optimizing protein purification in the effort to reach consistency and quality. Case studies will be presented that are based on science, yet illustrate the tenacity needed to maximize purity and yield, while minimizing purification steps. The agenda addresses process development and issues of scale, along with purifying challenging proteins, such as membrane proteins.

TUESDAY, JANUARY 20


2:00 BuzZ Session A







PURIFYING MEMBRANE PROTEINS

8:15 Chairperson’s Opening RemarksWilliam Gillette, Ph.D., Senior Scientist, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research


8:20 The Distinct Issues of Membrane Proteins versus Soluble ProteinsRobert Stroud, Ph.D., Professor, Biochemistry/Biophysics and Pharmaceutical Chemistry and Director, Membrane Protein Expression Center, University of California at San Francisco (UCSF)Membrane proteins present distinct issues versus soluble proteins; pure, homogeneous and stable in solution. Eukaryotic proteins need expression in eukaryotic cells. These include yeast, and insect cells. Human proteins, to be expressed for antibody preparation, require proper tailoring and glycosylation; and can often be achieved only in human cells. Antibody partners can be the basis for proof of principle therapeutics, and for helping to purify, assay, and validate membrane proteins’ mechanisms.

9:00 Expression of Transmembrane Proteins in Yeast for Genetic, Biochemical and Structural StudiesMark E. Dumont, Ph.D., Professor, Biochemistry and Biophysics, University of Rochester Medical CenterThe difficulty of expression, solubilization, and purification of transmembrane proteins, particularly those from eukaryotes, constitutes a significant barrier to understanding their mechanisms. The use of a system based on expression of membrane proteins from various sources in baker’s yeast has allowed the use of genetic approaches for functional characterization and stabilization of expressed proteins, while facilitating purification of transmembrane enzymes, transporters, and receptors for biochemical studies and x-ray crystallographic structure determination.

9:30 Overproduction and Functional Characterization of Purified Equilibrative Nucleoside TransportersFranklin A. Hays, Ph.D., Assistant Professor, Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center The Solute Carrier Superfamily proteins constitute a functionally and structurally diverse family in eukaryotic membrane proteomes and are associated with modulating efficacy for an equally broad range of human therapeutics. Our recent focus is on defining the molecular basis for Equilibrative Nucleoside Transporter (ENT) inhibition, transport, and substrate recognition. ENTs have only been identified in eukaryotic organisms and have proven recalcitrant to overexpression and functional characterization in purified form - this despite the fact that they directly bind or transport over 25 FDA approved therapeutics. We have developed an optimized pipeline for the production of active, pure, eukaryotic ENTs and have used this system to obtain novel insights into the transport characteristics of the only identified ENT protein from Saccharomyces cerevisiae named Function Unknown Now 26 (FUN26). This talk will focus on our efforts to overproduce, purify, solubilize, and reconstitute functional ENT proteins along with novel insights into ENT transport properties.


10:50 GPCR Purification: Protein Engineering and Process Optimisation are Key Elements for Successful GPCR CrystallisationMarkus Koglin, Ph.D., Associate Director, Protein Engineering, Heptares TherapeuticsThe generation of Stabilised G-protein Coupled Receptors (StaRs) has opened new routes in the purification procedures for GPCRs. Here we demonstrate that protein quality is drastically improved with increasing thermostability. StaR generation alone normally does not provide suitable material for crystallisation. The combination of StaR technology, systematic construct design, and careful optimisation of the purification process delivers a powerful approach to generate protein with suitable crystallisation qualities.






JANUARY 21-22





























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11:20 Strategies for Detergent Stabilization and Large-Scale Affinity Purification of the Functional G Protein-Coupled ReceptorsAlexei Yeliseev, Ph.D., Staff Scientist, Group Leader, LMBB, NIAAA, National Institutes of Health (NIH)Human cannabinoid receptor CB2, a G protein-coupled receptor involved in regulation of immune response, is an important target for pharmaceutical drug development. For high resolution structural studies, milligram quantities of pure, homogenous and functional protein are required. We expressed the functional CB2 receptor in E. coli, and optimized its purification by tandem affinity chromatography. An efficient strategy for stabilization of the receptor in detergent micelles and reconstituted into lipid bilayers was developed.

11:50 Overcoming Difficult to Express Proteins: Cell-Free Additives for Solubilizing and Characterizing Membrane ProteinsMatthew Coleman, Ph.D., Deputy Group Leader, Molecular Toxicology, Lawrence Livermore National Laboratory; Senior Scientist, University of California at DavisWe have developed cell-free methods for producing membrane proteins, which are inherently difficult to obtain. These include nanolipoprotein particles (NLPs), telodendrimers, which can be combined with NLPs or lipids, and amphipathic peptides. I will further discuss how we use the three different additives and their ability to support membrane protein solubility as well as function. These processes are easily adapted to high-throughput technologies for feeding the membrane structural biology pipeline.

12:20 High Throughput Strategies for MAB and FAB Development

Sponsored by

Catherine Allioux, Global Product Manager, Chromatography Sorbents and Membranes, Pall Life SciencesDeveloping a purification process using conventional methods is less and less compatible with biopharm industry challenges in terms of timelines and cost constraints. High Throughput Process Development (HTPD) for screening chromatography sorbents and multiple process conditions, based on a Design of Experiment (DoE) approach, has become a standard that enables saving time and sample, while improving process efficiency. We discuss mAb and Fab purification strategies leading to a three step process set up within 12 days.

12:50 Session Break

1:00 Luncheon Presentation I: Accurately Assessing Comparibility of Biosimilar Interferon From Refolding to Polishing During Process Development

Sponsored by

Paul Belcher, Ph.D., Market Development Leader, GE Healthcare Life SciencesDevelopers of biosimilars use an array of biomolecular characterization methods to prove that their processes are well understood, robust, and controlled. Extensive analytical studies including comparative physicochemical and functional studies are needed to confirm similarity. This talk will describe a process for HTPD of IFNα-2a, compared throughout the development process with a market available reference molecule. The focus is on the use of Biacore™ T200 and Amersham™ WB system for accurate comparability studies during this development.

PURIFYING ANTIBODIES2:00 Chairperson’s RemarksMatthew Coleman, Ph.D., Deputy Group Leader, Molecular Toxicology, Lawrence Livermore National Laboratory, and Senior Scientist, University of California at Davis

2:05 Resolving Protein Purification Challenges for Therapeutic Antibody DevelopmentErin Christensen, MBA, Research Associate, Protein Chemistry, Genentech, Inc. -- – A Member of the Roche GroupPurification technology is fundamental in antibody drug discovery. Maintaining biological and biophysical features of antigens is critical to generate antibodies with high diversity and specificity. Various antigen forms and orthologs are required for antibody screening and cross-species affinity. With increasing numbers of new formats of antibody fragments as therapeutic proteins, there are more technical demands in purification and characterization. This talk will present case examples in dealing with target-specific challenges.

2:35 Development of an Industrially Relevant DSP Platform Process for a Bispecific Antibody, the κλ-BodyJean-François Depoisier, Head, Downstream Processing Unit, NovImmune SAIn order to exploit novel mechanisms of action and achieve superior clinical efficacy, NovImmune has developed a novel bispecific antibody format, the κλ-body that has a molecular structure identical to a fully human monoclonal antibodies. The talk will address the challenges in developing an industrially relevant purification process for this innovative bispecific antibody format. The strategy for identifying optimal work space based on a broad range scanning of process parameters and novel purification technologies will be presented.

3:05 BEAT Bispecific Antibody: Development of a Process with a Platform ApproachSonia Letestu, Process Development Engineer, DSP, Glenmark Pharmaceuticals S.A.The BEAT® is a novel bispecific antibody techonology developed by Glenmark Pharmaceuticals. The scFv-FAB BEAT® format is a combination of a Fab arm and an ScFv arm with a fully functional Fc part, conserving key IgG properties such as thermostability, the potential for effector function and antibody-like pharmacokinetics. The presentation will present a new Bispecific technology specically engineered in order to allow an efficient impurity removal in one single step.

3:35 A New Alkali Stable Adsorbent for Antibody Purification

Sponsored by

Steve Burton, Ph.D., CEO, ProMetic BioSciences Ltd. A small synthetic ligand that is alkaline stable has been designed and developed for the affinity purification of monoclonal antibodies. Data presented will demonstrate that the adsorbent, comprised of a synthetic ligand immobilised onto an agarose base matrix, performs comparably to the leading Protein A products in terms of binding capacity, recovery and purity.






JANUARY 21-22





























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3:50 A Step Change in Protein Purification Sponsored by

Michael Bavand, Ph.D., CEO, ChromaCon AG Twin-column protein purification for mAb platform processes enable significant cost savings and increased processing speed. Contichrom CUBE equipment is designed as a modular lab-scale system to run batch, cyclic and continuous processes that can be seamlessly scaled up to GMP pilot /process scale using LEWA’s EcoPrime Twin LPLC system.






VACCINE PURIFICATION

8:30 Chairperson’s RemarksMarkus Koglin, Ph.D., Associate Director, Protein Engineering, Heptares Therapeutics

8:35 Purification of the Sanofi Pasteur HSV2 Vaccine Candidate, HSV529Sophia Mundle, Ph.D., Deputy Director, Protein Chemistry, Sanofi Pasteur Biologics Co.The Sanofi Pasteur replication defective HSV2 vaccine candidate, HSV529, can be purified by a method which includes a combination of harvesting without cell disruption, endonuclease treatment, depth filtration, anion-exchange chromatography and ultrafiltration/diafiltration (UF/DF). The resultant virus retains infectivity and is ∼200-fold more pure with respect to host cell DNA and proteins than is HSV529 purified by ultracentrifugation. Side-by-side comparison of chromatography-purified ACAM529 with sucrose cushion-purified HSV529 shows that both preparations are equally immunogenic and protective when tested in vivo.

9:05 Structural Characterization of Complex Glycoconjugate VaccinesCarlo Zambonelli, Ph.D., Principal Scientist, Biophysics, Novartis VaccinesA quality-by-design approach and increasing requirements from regulatory agencies demand an ever-increasing structural characterization of complex antigens such as glycoconjugates, formylated proteins and phospholipid vesicles. We are focusing on two complementary directions: 1) characterization of antigens in complex mixtures and 2) fine structural characterization of complex antigens. As an example, advanced applications of SEC-MALS and LC/MS to the structural characterization of glycoconjugates will be discussed.



OVERCOMING PURIFICATION CHALLENGES

10:50 Reproducing Irreproducible Results: A Case Study for Robust Kinase CrystallizationMario Lebendiker, Ph.D., Head, Protein Purification Facility, Hebrew University of JerusalemI will present a case study for robust production of an important human Kinase, with possible therapeutic implications. The reproducible production of different batches in E.coli, allows us to successfully crystallize our target with a great variety of ligands. We describe our expression and purification approach, bottlenecks, etc. Since this protein was crystallized in the past, this case study emphasizes the necessity to establish “minimal protein quality information” in publications in order to assure reproducibility of results.

11:20 Production of Prenylated Proteins in the BEVS: Purification and AnalysisWilliam Gillette, Ph.D., Senior Scientist, Cancer Research Technology Program, Frederick National Laboratory for Cancer ResearchProduction and isolation of recombinant prenylated proteins has been problematic due to the heterogeneous nature of the post-translational processing. Many proteins that are considered important by pharmaceutical companies are prenylated in vivo. The isolation of these molecules in a homogenous state has proven challenging. We will present our progress in addressing these issues using the BEVS. Data will include expression, purification, and molecular characterization.

11:50 Osmotic Virial Coefficients as Access to the Protein Partitioning in Aqueous Two-Phase ExtractionChristoph Brandenbusch, Ph.D., Group Leader, Bioprocess Separations, Laboratory of Thermodynamics, Biochemical and Chemical Engineering, TU DortmundThe selection of Aqueous Two-Phase Extraction (ATPE) systems for protein purification in state-of-the-art process development is often based on excessive screening (phase forming components, displacement agents, pH, and temperature). This is time and cost intensive and thus, although highly efficient, makes ATPE systems rather unattractive for industrial applications. By using a thermodynamics based approach, it is possible to dramatically simplify the ATPE system selection using static and dynamic light scattering experiments.







4th Annual Higher-Throughput Protein Purification

Supporting Technologies

JANUARY 22-23





























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High-throughput processes have transformed the traditional protein-by-protein trial-and-error approach for testing criteria and scaling up. This leading meeting on Higher-Throughput Protein Purification investigates HTP in the quest to develop methods that ensure quality and translate to large scale. Automation, robotics and liquid handlers will be discussed, along with developing small-scale models and HTP screening. Case studies illustrate how leaders in the field are integrating HTP approaches in order to reduce the time and effort needed to successfully establish parameters and achieve pure protein.





HTP AUTOMATION

2:00 Chairperson’s Opening RemarksOpher Gileadi, Ph.D., Principal Investigator, Genome Integrity and Repair, The Structural Genomics Consortium (SGC), University of Oxford


2:05 Automated Higher-Throughput Protein Purification Using Enhanced ÄKTA Chromatography InstrumentsKenneth Walker, Ph.D., Scientific Director, Biologics, Amgen, Inc.In order to keep pace with the increase in high-throughput cloning and expression needed to screen large panels of therapeutic candidates, we developed systems to substantially enhance the throughput of ÄKTA chromatography instruments through the use of novel large format autosamplers and quasi-parallel processing techniques. With little user intervention, these instruments can process a wide array of samples employing flexible, tandem, two column processing rapidly generating high quality products.

2:45 Acceleration of Purification Process Development Using High-Throughput AutomationSrinivas Chollangi, Ph.D., Scientist, Bristol-Myers Squibb Co.By incorporating high-throughput (HT) technologies at key points in development, we show that an operating space for in-process conditions can be defined quickly and improve product and process knowledge prior to scale-up experiments. This allows for rapid process development of mAbs while minimizing the material requirement for early-stage development. As HT technologies continue to emerge, this approach will become increasingly powerful and will be applied to downstream process development of other biologics.

3:15 Automation of Non-Chromatographic Downstream Processes Gaining on High-Throughput Bioprocess DevelopmentCornelia Walther, Ph.D., Scientist, Applied Microbiology, University of Natural Resources and Life Sciences ViennaWe present an automated platform for parallel screening of inclusion body solubilization and protein refolding conditions using DoE. The incompatibility of numerous compounds in refolding buffers is overcome by elution of the captured protein in a single buffer. This platform enables the screening for optimized conditions of the entire protein recovery from inclusion bodies creating a holistic view on all crucial impact factors in an early stage of process development.



HIGH-THROUGHPUT TECHNIQUES TO ENSURE QUALITY

5:00 Using High-Throughput Techniques to Produce Difficult Targets: Flavivirus NS1, a Case StudyWilliam (Clay) Brown, Ph.D., Associate Research Scientist, Life Sciences Institute, University of MichiganFlavivirus NS1 protein is a multi-functional protein that is required for genome replication and is involved in immune system evasion. These key roles in the Flavivirus infection cycle make NS1 an attractive target for drug and/or vaccine development. We applied high-throughput cloning and expression evaluation techniques for the identification of constructs that would be suitable for crystallization and 3-D structure determination. We also used a matrix buffer screen for initial purification development.

5:30 Q & A with Speakers







JANUARY 22-23





























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FRIDAY, JANUARY 23


CONSTRUCTING AND INTEGRATING AN HTP PROTEIN PURIFICATION PLATFORM

8:30 Chairperson’s RemarksSrinivas Chollangi, Ph.D., Scientist, Bristol-Myers Squibb Co.

8:35 Building a High-Throughput Protein Purification PlatformJiansheng Wu, Ph.D., Senior Scientist, Protein Chemistry, Genentech, Inc. – A Member of the Roche Group Over many years, we have built a robust high-throughput protein purification platform for most commonly used tags. For this tagged proteins, we can combine NiNTA with different secondary columns, such as SEC and anti-Flag. We also have a GST-SEC two step purification for GST tagged proteins. In addition, we also developed a prep-grade autosampler to complement these methods. Thousands of proteins have been purified with the high throughput platform.

9:05 High-Throughput Strategies to Rescue Protein Expression and Solubility: Perspectives from a Protein Expression and Purification PipelineStephen Nakazawa Hewitt, Ph.D., Research Scientist II, University of Washington School of MedicineInsoluble and non-expressing recombinant constructs are major obstacles for purification. Using methods developed from structural genomics efforts we have designed a number of strategies for rescue of recalcitrant constructs. A customized panel of small molecules added during cell lysis and a fusion vector consisting of the highly soluble maltose binding protein allows for rescue attempts of previously insoluble recombinant proteins expressed in E. coli.

9:35 Application of Modular Expression Toolboxes in High-Throughput Protein ExpressionErnst Weber, Ph.D., Lab Head, Protein Engineering and Assays, Bayer HealthCareRapid, parallel expression optimization is essential in drug development to ensure production of high quality target proteins. Therefore we set up a modular expression toolbox, consisting of standardized elements influencing expression levels, which allow the generation of complex expression libraries. Here we will discuss advantages and implications of a modular cloning system, outline the strategy and design of an expression toolbox, and present case studies detailing how such libraries were expressed and analyzed.


HTP SCREENING11:00 Simulation of Diafiltration Incompatibilities via a High-Throughput Solubility ScreenGregory Barker, Ph.D., Senior Research Investigator, Biologics Process Development, Bristol-Myers Squibb Co.Biologics manufacturing processes usually end with a tangential flow filtration (TFF) step to concentrate and diafilter the unformulated drug substance. Design space mapping is critical yet hampered by the lack of high-throughput screening (HTS) tools. One major roadblock to HTS is the microscaling of TFF fluid mechanics. A high-throughput solubility screen combined with shear susceptibility assessment is proposed here to simulate and predict a favorable diafiltration design space.

11:30 SEC-TID, a Small Molecule Target ID Platform Utilizing a PDB-Guided Small Molecule-Ligandable Protein LibraryAntonin Tutter, Ph.D., Scientific Technical Leader I, Novartis Institutes for BioMedical Research, Inc.Size Exclusion Chromatography for Target ID (SEC-TID) is a novel high-throughput protein:small molecule target identification platform that utilizes a library of thousands of individually expressed and purified human-ligandable protein domains to identify small-molecule binding interactions. We describe SEC-TID, as well as the PDB-guided bioinformatics approach and high-throughput PIPE cloning workflow used to generate the clones and purified proteins that comprise the SEC-TID protein library.

12:00 pm High-Throughput Screening as a Tool for Developing Scalable Purification Processes of Biotherapeutic ProteinsManfred Saller, Ph.D., Research Scientist, Downstream Processing, Synthon B.V.Development of purification processes has become in recent years a more structure-based approach by using high throughput screening technologies and statistical design. Different groups explored a variety of conditions at microscale format developing more robust purification processes for biologics with low product amounts. One of the challenges in these developments concerns the translation of purification processes from microscale format in microtiter plates towards a lab scale format in columns, which will be explained in this presentation.


1:00 Session Break


HTP PURIFICATION TO ENABLE DRUG DISCOVERY2:00 Chairperson’s RemarksPauline M. Rudd, Ph.D., Professor, Glycobiology, National Institute for Bioprocessing Research and Training (NIBRT)






JANUARY 22-23

2:05 High-Throughput Purification of Antibodies and Antibody Fragments Enabling Biologic Drug DiscoveryBenjamin Kemp, Ph.D., Senior R&D Manager, MedImmune, LLCMedImmune research employs several strategies to enable the purification of antibodies and antibody fragments. These encompass both high-throughput 96-well plate based technologies and methods to generate milligram quantities of hundreds of antibodies per week. Purified antibodies are used in functional screening enabling lead identification and characterisation.

2:35 A Scaled-Down Scale-Down Model: HTPD Case StudiesDouglas MacDonald, Senior Scientist, Purification & Conjugation Development, Seattle GeneticsDue to increasingly compressed drug development timelines, companies are leveraging robotics/high throughput techniques to scale-down, automate and therefore accelerate many aspects of purification and analytical development. However, careful consideration needs to be used in achieving comparable results to established small-scale models. The goal of this application was to utilize 200 and 600μL chromatography columns eight at a time, on a liquid handling system in two ways. First, to develop a scaled-down Protein A model for use in upstream and downstream process development, and second, to augment or even replace a resin-tip based analytical sample prep assay to achieve comparable results to the platform purification process.

3:05 High-Throughput Protein Engineering for Optimal Purification and CrystallizationOpher Gileadi, Ph.D., Principal Investigator, Genome Integrity and Repair, The Structural Genomics Consortium (SGC), University of OxfordThe SGC has purified more than 2000 human proteins for crystallization and biochemical studies, with a success rate of ~50%. Soluble expression was achieved by testing multiple truncation constructs, expression vectors and hosts. We are now implementing both scanning and directed mutagenesis techniques to enable production of more difficult proteins. Prioritizing mutation sites allows us to achieve this with a modest level of mutagenesis.

3:35 A High-Throughput Enabled Pipeline for Eukaryotic Expression and PurificationRonald D. Seidel, III, Ph.D., Director, Macromolecular Therapeutic Development, and Associate Director, Albert Einstein Protein Production Facility, Biochemistry, Albert Einstein College of MedicineHigh-throughput approaches enable more to be done with limited resources. Here, we have developed high throughput methodologies to produce eukaryotic target proteins in eukaryotic hosts; we have sought to use integrated ‘off the shelf’ equipment and to generate protocols that could also be replicated in a low automation environment that could increase the productivity of any laboratory seeking to produce proteins in insect or mammalian cell hosts.


Student FellowshipProgram

Present Your Poster to 1,200+ Protein Science ResearchersStudent Fellowship Award Winners will attend the 14th Annual PepTalk: The Protein Science Week for as low as $295*

Full-time graduate students and Ph.D. candidates are encouraged to apply for the PepTalk 2015 Student Fellowship. Twenty fellowship award winners will receive a poster presentation slot and a savings of over $900 on their registration fee. Applications are due by October 24, 2014.

STUDENT FELLOWSHIP DETAILS:

• Full-time graduate students and Ph.D. candidates are eligible to apply for the PepTalk Student Fellowship. Applications are due by October 24, 2014.

• Interested students must complete the 2015 Student Fellowship Application and are required to present a scientific poster (title and abstract are due at the time of the application).

• All applications will be reviewed by the scientific review committee and accepted students will be notified no later than November 5, 2014 if they have been accepted for the 2015 Student Fellowship.

• Accepted 2015 Student Fellows will receive a discounted conference registration rate of $295*, which must be paid by November 21.

• This fellowship is limited to 20 students and is for the Premium Conference Package Only. Excludes Short Courses.

• All accepted 2015 Student Fellows will be asked to help promote the conference at their college and throughout their social media networks.

• Students not accepted for the 2015 Student Fellowship can still register at a discounted rate of $595*, and will not be required to present a poster.

• ADDED BONUS! All recipients will be entered in the Poster competition featuring cash prize winners.

* This discounted rate cannot be combined with any other discounts for this event. Your discounted registration does not grant access to any of the short courses or pre-conference events. It also does not include hotel, travel or meals.





























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ACCOMPANYING CONFERENCE: MEMBRANE PROTEINS

A Valuable Resource and Target

JANUARY 21-22





























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Membrane proteins, the gateways to the cell, are valuable drug targets. To design better-targeted drugs, researchers must know their structure and functional characteristics. But determining them is difficult because of the large amounts of membrane protein needed. Sources are scarce due to the practical problems of working with membrane proteins in expression, purification and crystallization. The Membrane Proteins conference addresses strategies and solutions for their extraction, expression and purification, and with case studies showcasing their value as an antibody drug target. Explore how to obtain functional membrane proteins and learn more about this important protein class.

TUESDAY, JANUARY 20


2:00 BuzZ Session A







PURIFICATION

8:15 Chairperson’s Opening RemarksWilliam Gillette, Ph.D., Senior Scientist, Cancer Research Technology Program, Frederick National Laboratory for Cancer Research


8:20 The Distinct Issues of Membrane Proteins versus Soluble ProteinsRobert Stroud, Ph.D., Professor, Biochemistry/Biophysics and Pharmaceutical Chemistry; Director, Membrane Protein Expression Center, University of California at San Francisco (UCSF)Membrane proteins present distinct issues versus soluble proteins; pure, homogeneous and stable in solution. Eukaryotic proteins need expression in eukaryotic cells. These include yeast, and insect cells. Human proteins, to be expressed for antibody preparation, require proper tailoring and glycosylation; and can often be achieved only in human cells. Antibody partners can be the basis for proof of principle therapeutics, and for helping to purify, assay, and validate membrane proteins’ mechanisms.

9:00 Expression of Transmembrane Proteins in Yeast for Genetic, Biochemical and Structural StudiesMark E. Dumont, Ph.D., Professor, Biochemistry and Biophysics, University of Rochester Medical CenterThe difficulty of expression, solubilization, and purification of transmembrane proteins, particularly those from eukaryotes, constitutes a significant barrier to understanding their mechanisms. The use of a system based on expression of membrane proteins from various sources in baker’s yeast has allowed the use of genetic approaches for functional characterization and stabilization of expressed proteins, while facilitating purification of transmembrane enzymes, transporters, and receptors for biochemical studies and x-ray crystallographic structure determination.

9:30 A Novel Strategy to Express and Purify Highly Stable and Functional GPCR Signaling ComplexesArun Shukla, Ph.D., Assistant Professor, Medicine, Duke UniversityStructural characterization of GPCR signaling complexes remains a very challenging but extremely important task. Molecular visualization not only reveals fundamental mechanism of assembly and functioning of such complexes but also provides a unique framework for drug discovery. We present a novel strategy to assemble a highly stable and functionally competent GPCR signaling complex in-cellulo and its subsequent purification for direct structural characterization.


10:50 GPCR Purification: Protein Engineering and Process Optimisation are Key Elements for Successful GPCR CrystallisationMarkus Koglin, Ph.D., Associate Director, Protein Engineering, Heptares TherapeuticsThe generation of Stabilised G-protein Coupled Receptors (StaRs) has opened new routes in the purification procedures for GPCRs. Here we demonstrate that protein quality is drastically improved with increasing thermostability. StaR generation alone normally does not provide suitable material for crystallisation. The combination of StaR technology, systematic construct design, and careful optimisation of the purification process delivers a powerful approach to generate protein with suitable crystallisation qualities.





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11:20 Strategies for Detergent Stabilization and Large-Scale Affinity Purification of the Functional G Protein-Coupled ReceptorsAlexei Yeliseev, Ph.D., Staff Scientist, Group Leader, LMBB, NIAAA, National Institutes of Health (NIH)Human cannabinoid receptor CB2, a G protein-coupled receptor involved in regulation of immune response, is an important target for pharmaceutical drug development. For high resolution structural studies, milligram quantities of pure, homogenous and functional protein are required. We expressed the functional CB2 receptor in E. coli, and optimized its purification by tandem affinity chromatography. An efficient strategy for stabilization of the receptor in detergent micelles and reconstituted into lipid bilayers was developed.

11:50 Overcoming Difficult to Express Proteins: Cell-Free Additives for Solubilizing and Characterizing Membrane ProteinsMatthew Coleman, Ph.D., Deputy Group Leader, Molecular Toxicology, Lawrence Livermore National Laboratory; Senior Scientist, University of California at DavisWe have developed Cell-free methods for producing membrane proteins, which are inherently difficult to obtain. These include nanolipoprotein particles (NLPs), telodendrimers, which can be combined with NLPs or lipids, and amphipathic peptides. I will further discuss how we use the three different additives and their ability to support membrane protein solubility as well as function. These processes are easily adapted to high-throughput technologies for feeding the membrane structural biology pipeline.

12:20 High Throughput Strategies for MAB and FAB Development

Sponsored by

Catherine Allioux, Global Product Manager, Chromatography Sorbents and Membranes, Pall Life SciencesDeveloping a purification process using conventional methods is less and less compatible with biopharm industry challenges in terms of timelines and cost constraints. High Throughput Process Development (HTPD) for screening chromatography sorbents and multiple process conditions, based on a Design of Experiment (DoE) approach, has become a standard that enables saving time and sample, while improving process efficiency. We discuss mAb and Fab purification strategies leading to a three step process set up within 12 days.

12:50 Session Break

1:00 Luncheon Presentation : Accurately Assessing Comparibility of Biosimilar Interferon From Refolding to Polishing During Process Development

Sponsored by

Paul Belcher, Ph.D., Market Development Leader, GE Healthcare Life SciencesDevelopers of biosimilars use an array of biomolecular characterization methods to prove that their processes are well understood, robust, and controlled. Extensive analytical studies including comparative physicochemical and functional studies are needed to confirm similarity. This talk will describe a process for HTPD of IFNα-2a, compared throughout the development process with a market available reference molecule. The focus is on the use of Biacore™ T200 and Amersham™ WB system for accurate comparability studies during this development.

DIFFICULT EXPRESSION AND PRODUCTION

2:00 Chairperson’s RemarksIan Hunt, Ph.D., Group Leader, Proteomic Chemistry and Head, Protein Sciences, Novartis

2:05 Designer Surfactant-Like Self-Assembling Peptides for Membrane Protein Purification and StabilizationSotirios Koutsopoulos, Ph.D., Research Scientist, Center for Biomedical Engineering, Massachusetts Institute of TechnologyMembrane proteins are integral proteins of the cell membrane and are directly involved in the regulation of many biological functions and in drug targeting. However, our knowledge of membrane proteins is limited due to difficulties in producing sufficient quantities of soluble, functional and stable receptors. Designer, surfactant-like peptides may be used to extract the protein from the cell membrane and stabilize the protein outside the membrane bilayer for further studies.

2:35 Production of Human Integral Membrane Proteins in Mammalian CellsJames D. Love, Ph.D., Director, Technology Development, Biochemistry, Albert Einstein College of MedicineIntegral membrane proteins are key targets in the understanding of health and human disease, yet producing functional material in great enough quantities for structural studies remains a formidable task. This talk highlights the expression technologies under development that will greatly aid high-throughput efforts to produce functional human membrane proteins and complexes, specifically GPCRs, for a plethora of structural techniques.

3:05 Advances for Maximizing Protein Yields in HEK293 and CHO Transient Expression

Sponsored by

Henry C. Chiou, Ph.D., Associate Director, Cell Biology, Life Science Solutions, Thermo Fisher ScientificIncreasing focus on complex proteins in advanced research elevates the needfor higher yields from mammalian transient expression systems. To achievethis objective while maintaining speed, simplicity and ease-of-use requirescoordinated development and synergy between novel high-capacity media,higher productivity HEK293 and CHO cells, and higher performance transfectionreagents. This presentation will demonstrate the effectiveness of this “holistic”approach to achieve gram-level protein yields by transient expression.

3:35 The Need for Technology Platforms in USP Development

Sponsored by

Hugo de Wit, CEO, CellcaAre you faced with budget reductions for USP development or even anticipated to do “more with less”? Do you see increasing expectations from market and management regarding reducing timelines and cost of goods attributes? We demonstrate how a robust USP platform can help you meet these challenges.

3:50 Bypassing Inclusion Body Formation in E. coli:



Maximizing Soluble Expression of Complex Multimeric Proteins

Sponsored by

Mark Valasek, M.D., Ph.D., Co-founder and Scientific Director, AbSciCytoplasmic expression of large complex proteins in E. coli is limited primarily due to protein aggregation and the corresponding refolding process. These costly aggregates form when proteins are expressed too quickly and strongly. AbSci has developed a tightly regulated, dual titratable expression system that is able to homogenously induce high levels of complex multimeric proteins in a soluble and active form.






DISCOVERY AND DEVELOPMENT OF ANTIBODY TARGETS

8:30 Chairperson’s RemarksTrevor Wilkinson, Ph.D., Associate Director, Protein Sciences, Antibody Discovery and Protein Engineering, MedImmune

8:35 GPCR Expression by Individual Cell Types: Novel Membrane Targets for Therapeutic AntibodiesPaul Insel, Ph.D., Professor, Pharmacology & Medicine, University of California, San DiegoGPCRs, the largest family of membrane receptors, also represent the largest class of targets of FDA-approved drugs. However, little is known regarding GPCR expression by individual cell types. Using a GPCRomic strategy, we have identified the full range of non-chemosensory GPCRs, including numerous orphan GPCRs, expressed by many such cell types. Our other findings suggest the possibility of using antibody therapeutics directed at such receptors.

9:05 Targeting T Cells with an Anti-Ion Channel Antibody with Ultralong CDR H3sVaughn Smider, M.D., Ph.D., Assistant Professor, Molecular Biology, The Scripps Research InstituteThe relatively flat binding surface of a typical antibody paratope may not allow optimal interactions with certain epitopes. Cow antibodies contain ultralong CDR H3’s consisting of a b-ribbon stalk and disulfide-bonded knob. The knob structures are reminiscent of ion channel bioactive peptides known to interact with high specificity and affinity with ion channels. We have engineered a

cow antibody to bind and inhibit an ion channel critical for T-cell activation in autoimmune disease and inflammation.



10:50 Strategies to Obtain Antibodies to Difficult Membrane Protein TargetsJohn S. Kenney, Ph.D. President & CEO, Antibody Solutions Some membrane protein targets, including high homology proteins, G-protein-coupled-receptors (GPCRs), Ion Channels, and Multicomponent Receptor complexes present unique challenges in the generation of specific, high-affinity, and functional antibodies. Each target requires a custom approach based upon the nature of the target and the desired characteristics of the antibody. Strategies for antigen design, immunization, and screening of antibodies to difficult membrane proteins and results obtained will be presented.

11:20 Discovery and Optimization of Novel Anti G-Protein Coupled Receptor Monoclonal AntibodiesTrevor Wilkinson, Ph.D., Associate Director, Protein Sciences, Antibody Discovery and Protein Engineering, MedImmuneG-protein coupled receptors represent a challenging target class for the isolation and optimization of therapeutic biologics. We have used a combination of immunization and phage display to isolate functional antagonistic antibodies targeting a chemokine receptor and a formyl peptide receptor that will be presented as case studies. We also describe how combinatorial mutagenesis approaches have been used to make significant improvements to both affinity and species cross-reactivity of a lead molecule and demonstrate that the optimised antibodies show significantly increased potency in cellular disease assays.

11:50 High-Throughput Strategies to Obtain High Affinity, Specific, and Conformationally Selective Recombinant Antibodies to Membrane Proteins by Phage DisplayMarcin Paduch, Ph.D., Technical Director, Synthetic Antibody & Crystallography Core Facility, The University of ChicagoState of the art methods for generating recombinant antibodies to membrane proteins require the use of detergents that do not necessarily mimic the native lipid environment. We have developed a suite of next generation high-throughput technologies to generate high affinity, specific, and conformationally selective reagents by exploiting liposomes, nanodiscs and cell surface display for antigen presentation. These native-like environments create the possibility of trapping physiological states otherwise not accessible by current methods.






JANUARY 21-22





























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ACCOMPANYING CONFERENCE: CHO CELLS

Engineering the Process from -Omics to Production





























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To meet the huge demand for recombinant proteins, advances in CHO cell culture technology continue to significantly improve production. This achievement is due to progress in both engineering stable and transient cell lines as well as optimizing the cell culture process. When both are accomplished, higher-production titers and better product quality result. The CHO Cells conference gathers cell culture specialists and cell line engineers to explore the latest data, tools and strategies for improving biotherapeutic protein production, expression and product quality.



IMPROVING PRODUCTION AND PRODUCT QUALITY

2:00 Chairperson’s RemarksJie Chen, Ph.D., Professor, Electrical and Chemical Engineering, University of Alberta, Edmonton

»FEATURED PRESENTATION 2:05 A Novel Approach to Controlling Cell Growth and Improving Specific Productivity and Product Quality in CHO Cell CulturesZhimei Du, Ph.D., Principal Scientist, Cell Culture Development & Manufacturing, Teva Pharmaceuticals USAWe identified a novel approach which can control cell growth and improve specific productivity, as well as product quality simultaneously. Results demonstrate that the cell proliferation is consistently controlled in all recombinant cell lines throughout the production processes with specific productivities increased to as much as 110 pg/cell/day. Additionally, glycan processing is improved by decreasing high mannose and increasing mature G1 and G2 glycoforms. Molecular mechanisms of the related signaling pathways have been studied and will be discussed.

2:35 Increasing Monoclonal Antibody Production Using Low-Intensity Pulsed UltrasoundJie Chen, Ph.D., Professor, Electrical and Computer Engineering, University of Alberta, EdmontonThe clinical and commercial success of monoclonal antibodies has led to the need for large-scale production in mammalian cell culture. Our low-intensity pulsed ultrasound (the claims of U.S. patent were allowed) increases production of antibodies in both hybridomas and CHO cells by stimulating the cells. Up to 60% increase in antibody production in hybridoma cells, and up to 25% increase in CHO cells have been achieved within a few days of stimulation.

3:05 Development of Innovative Processes for Expression of Monoclonal Antibodies

Sponsored by

Amita Goel, MS, CEO, CelltheonRobust and reproducible processes are required to generate high quality monoclonal antibodies that can accelerate into the clinic. To meet these demands Celltheon has developed CHO based SMART Expression System and SMART Bioprocessing Technology ™ that has generated high producing stable GMP manufacturing cell lines and processes that increase titers ~6-20 fold, increase yield, product quality and reduce time lines.

3:35 A Stable Episomal Expression System for Protein Production

Sponsored by

Meelis Kadaja, Ph.D., CSO, Icosagen Cell FactoryWe have developed a novel technology to stably maintain expression vectors in dividing mammalian cells. This enables the quick production of recombinant proteins in gram quantities as well as the ability to tailor the system to improve the quality of each protein. Our system is scalable and suitable for cell bank generation and recombinant antibody production.






CELL LINE ENGINEERING AND DEVELOPMENT

8:30 Chairperson’s RemarksHelene Faustrup Kildegaard, Ph.D., Co-Principal Investigator, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark

8:35 Omics-Guided Cell Line Engineering: Reducing High Mannose by Overexpressing N-Glycosylation Pathway RegulatorsShivani Gupta, Associate Scientist, Cell Line Development, Amgen, Inc.High mannose is a critical quality attribute for recombinant therapeutic monoclonal antibodies that can impact biological activity by influencing Fc-mediated effector functions, product stability, clearance rate and safety. In this study, we identified key genes whose expression is correlated with lower levels of high-mannose glycans in antibody-producing CHO cell lines.

9:05 Improving CHO Cell Factories Using Systems Biology and Genome Editing TechnologiesHelene Faustrup Kildegaard, Ph.D., Co-Principal Investigator, Novo Nordisk Foundation Center for Biosustainability, Technical University of DenmarkNew opportunities are arising to improve CHO cell factories using emerging systems, biology data and efficient genome editing technologies. Here, our recent efforts in improving CHO cell factories by genetic engineering using mainly genomics and transcriptomics data combined with CRISPR Cas9 technology will be presented.

JANUARY 21-22





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9:35 ESETEC® 2.0: New Generation of E. coli Secretion Technology for the High-Yield Production of Fabs

Sponsored by

Andreas Anton, Ph.D., Head, Bioprocess Development, Wacker Biotech GmbHWACKER has profoundly refined its patented ESETEC® E. coli based system for the manufacture of biopharmaceuticals. Targeted genetic modifications and process optimization measures led to the development of new, extremely productive cell lines and fermentation procedures. ESETEC® 2.0 is now able to produce several grams per liter of secreted Fabs.


10:50 Identifying Targets for Engineering Protein Secretion in CHO Cells with Systems BiologyNathan E. Lewis, Ph.D., Assistant Professor, Lab of Systems Biochemistry and Cell Engineering, University of California, San DiegoOur recent whole-genome sequencing efforts for CHO have enabled the construction of systems biology models. We now use such models for detailed analysis of -omics data from CHO to deepen our understanding of differences between host cell lines and to develop predictions for enhancing cell growth, protein modification and protein secretion.

11:20 Overexpression of MicroRNAs Enhances Recombinant Protein Production in CHO CellsWan Ping Loh, Bioprocessing Technology Institute, Agency for Science, Technology and Research (A*STAR)Cell engineering using microRNAs (miRNAs) is an emerging strategy to increase recombinant protein production in mammalian cells. We identified miRNAs that were differentially expressed between high- and low-mAb-producing CHO cell clones. Stable overexpression of these miRNAs in a CHO-mAb cell line resulted in increased IgG production of ~30% in pools and up to 140% in clones, without major changes in product aggregation and N-glycosylation profile.

11:50 Use of CHO Cell Engineering for Improved Protein SecretionPierre-Alain Girod, Ph.D., CSO, SelexisTypically in mammalian cell-based production systems, there is an inverse relationship between cell proliferation rate and cell-specific recombinant protein production. Yet cellular resources can be redirected from cell biomass accumulation to recombinant protein synthesis by changing the physico-chemical environment of the cells. But folding and assembly are highly protein-specific. To increase the functional capability of the cells and thus volumetric product titer, we engineered the host cells by increasing the rate of protein folding and assembly.



Sponsored by


Low transfection efficiency of CHO cells is a major bottleneck hampering Transient Protein Production (TPP). Polyplus-transfection®, with its 10+ year

expertise in transfection, has developed a novel technologically advanced transfection solution specifically designed for bioproduction. FectoPRO™ outperforms currently available PEI-based and lipid-based transfection reagents. We will present data and protocols leading to unmatched protein and antibody yields in CHO and HEK-293 cells.


FLEXIBLE PRODUCTION WITH TRANSIENT CELL LINES

2:00 Chairperson’s Opening RemarksDominic Esposito, Ph.D., Director, Protein Expression Laboratory, Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc.


2:05 Application of UPR Response and Cell Cycle Checkpoint Engineering for CHO Cell Line ConstructionTakeshi Omasa, Ph.D., Professor, Institute of Technology and Science, University of TokushimaThe construction of high-producing CHO cells is an important issue for therapeutic protein production. We introduce our new approach for Unfolded Protein Response (UPR)-based cell line construction and accelerating transgene amplification system using cell cycle checkpoint engineering.

2:45 Protein Production by PEI-Mediated CHO TransfectionYves Durocher, Ph.D., Team Leader, Protein Production, Biologics and Subsequent Entry Biologics, Life Sciences - NRC Human Health Therapeutics Portfolio, National Research Council CanadaWe present data describing our CHO-EBNA1 transient expression platform for the production of various proteins. For more difficult-to-express proteins or proteins needed in large quantities, we also developed an inducible CHO pool platform that allows the generation of stable pools expressing high levels of monoclonal antibodies in less than 3 weeks post-transfection. This platform can also be used to derive stable CHO clones for manufacturing therapeutic r-proteins candidates.

3:15 Systems-Based Approaches for Enhancing Transient Protein Expression in CHO CellsJonathan Zmuda, Ph.D., Associate Director, R&D, Thermo Fisher ScientificCHO cells are known to express lower levels of transient proteins than HEK293, but are desirable for biopharmaceutical development since they are the most commonly used cell type for bioproduction. To address the need for a higher-expressing transient CHO platform, systems-based approaches were utilized whereby the latest advances in cell culture media and feeds, transfection reagents and enhancers were optimized through multifactorial DoE to generate a complete solution for enhanced protein expression with titers comparable to HEK293-based systems.



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FierceBiotechTHE BIOTECH INDUSTRY’S DAILY MONITOR

3:45 Rapid Protein Production with Flow Electroporation: Higher Titers, Cell Type Flexibility and Scalability

Sponsored by

James Brady, Ph.D., MBA, Director, Technical Applications, MaxCyte, Inc.As protein therapeutics grow, future success will depend on the ability to be flexible regarding cell type and the ability to quickly scale up or scale down. In this presentation, data from MaxCyte flow electroporation will be presented demonstrating its scalability, high transfection efficiency and cell viability of commonly used cells, including CHO-S, HEK293 and insect cells, and production of antibody titers >1 g/L in 2 weeks.


5:00 High-Throughput mAb Expression and Protein A Purification Platform Based on Transient CHOYashas Rajendra, Ph.D., Research Scientist, Biotechnology Discovery Research, Eli Lilly and CompanyWe developed a PEI-mediated transient CHO-GS KO expression system that can generate mAb titers up to 0.5 g/L. We also developed a semi-automated Protein A purification process. Using a single liquid handling robot, up to 72 unique mAbs can be simultaneously purified from 24DWP (deep-well plate) transfections. This process yields 0.3 mg-1 mg of high-quality mAb at concentrations >0.5 mg/ml in a standard formulation buffer, enabling rapid characterization of mABs in multiple assay formats.

5:30 PANEL DISCUSSION: CHO Cells: Transient, Stable or BothRecombinant protein expression in mammalian producer cells such as CHO offers significant advantages in protein quality and post-translational modification. Expression can be transient, which achieves fast process times and high yields, or stable, which takes greater time to convert but offers other gains. This panel convenes experts to share insights into the benefits and tradeoffs of meeting the demand for recombinant proteins by engineering transient or stable cell lines or a combination of both.Moderator: Dominic Esposito, Ph.D., Frederick National Laboratory for Cancer Research, Leidos Biomedical Research, Inc.Panelists: James Brady, Ph.D., MBA, MaxCyte, Inc.Yves Durocher, Ph.D., National Research Council CanadaTakeshi Omasa, Ph.D., University of TokushimaYashas Rajendra, Ph.D., Eli Lilly and CompanyJonathan Zmuda, Ph.D., Thermo Fisher Scientific





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http://www.ondrugdelivery.com/


Current Sponsors & Exhibitors (as of August 22, 2014)

CHI offers comprehensive sponsorship packages which include presentation opportunities, exhibit space, branding and networking with specific prospects. Sponsorship allows you to achieve your objectives before, during, and long after the event. Any sponsorship can be customized to meet your company’s needs and budget. Signing on early will allow you to maximize exposure to qualified decision-makers.

Podium Presentations — Available within Main Agenda!

Showcase your solutions to a guaranteed, targeted audience. Package includes a 15- or 30-minute podium presentation within the scientific agenda, exhibit space, on-site branding, access to cooperative marketing efforts by CHI, and more.

Luncheon PresentationsOpportunity includes a 30-minute podium presentation. Boxed lunches are delivered into the main session room, which guarantees audience attendance and participation. A limited number of presentations are available for sponsorship and they will sell out quickly. Sign on early to secure your talk!

Invitation-Only VIP Dinner/Hospitality Suite Sponsors will select their top prospects from the conference pre-registration list for an evening of networking at the hotel or at a choice local venue. CHI will extend invitations and deliver prospects, helping you to make the most out of this invaluable opportunity. Evening will be customized according to sponsor’s objectives i.e.:

• Purely social• Focus group• Reception style• Dinner with specific conversation focus

Exhibit Exhibitors will enjoy facilitated networking opportunities with qualified delegates. Speak face-to-face with prospective clients and showcase your latest product, service, or solution.Additional branding and sponsorship opportunities are available, including:

• Mobile App• Hotel Room Keys• Footprint Trails• Staircase Ads

• Literature Distribution (Tote Bag Insert or Chair Drop)

• Padfolios• Program Guide Advertisement

Looking for additional ways to drive leads to your sales team? Discover the difference by utilizing CHI’s database of over 800,000 life sciences & drug discovery professionals!CHI’s Lead Generation Programs will help you obtain more targeted, quality leads throughout the year. We will mine our database of 800,000+ life science professionals to your specific needs. We guarantee a minimum of 100 leads per program! Opportunities include:

• Whitepapers• Web Symposia• Custom Market Research Surveys• Podcasts

Advertising opportunities such as marketing and promotional emails are also available.

To customize your participation at this event, please contact:

Companies A - K:Jason GerardiManager, Business [email protected]

Companies L - Z:Carol DinersteinDirector, Business [email protected]• Ajinomoto Althea Inc.

• Aldevron

• Antibody Solutions

• Aragen Bioscience

• Aspen Research Corporation

• Avia Biosystems

• Bend Research

• Blue Sky BioServices

• Cellca GmbH

• Chemistry Research Solution, LLC

• Cobra Biologics

• DNA2.0, Inc.

• Entegris, Inc.

• Enzo Life Sciences

• FeF Chemicals

• ForteBio, A Division of Pall Life Sciences

• Freeslate, Inc.

• Fujifilm Diosynth Biotechnologies

• Genalyte, Inc.

• Genedata AG

• GenScript

• IMA Life North America

• KBI Biopharma

• Life Technologies Corporation

• Lonza

• Lucigen Corporation

• Lyophilization Technology

• Malvern Instruments, Inc.

• MaxCyte, Inc.

• Millrock Technology

• OmniSep – Diba Industries

• PBS Biotech

• PerkinElmer

• PhyNexus, Inc.

• Pfanstiehl, Inc.

• ProMetic Life Sciences Inc.

• Proteos, Inc.

• ProteinSimple

• Rainin Instrument, LLC.

• Rentschler Biotechnologie

• RheoSense, Inc.

• Saint-Gobain

• Selexis

• SensiQ Technologies

• Soluble Therapeutics

• SP Scientific

• Thomson Instrument Company

• Toxikon Corporation

• Trinean

• VTU Technology

• Wasatch Microfluidics, Inc.

• Wyatt Technology

• Xbrane Bioscience





























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BuzZ Sessions are facilitated, small-group discussions. Interactive participation leads to problem-solving solutions and future collaborations around focused topics.

If you have a topic idea or would like to moderate a table, please contact: Ann Nguyen at [email protected]

ALUMNI DISCOUNTCambridge Healthtech Institute (CHI) appreciates your past participation at our events. Through loyalty like yours, CHI has been able to build this event into a must-attend for senior-level decision makers.

As a result of the great loyalty you have shown us, we are pleased to extend to you the exclusive opportunity to save an additional 20% off the registration rate.

Just check off the box marked Alumni Discount on the registration form to receive the discount!

Please note: Our records must indicate you were an attendee at a past CHI event to qualify.

The Intro-Net offers you the opportunity to set up meetings with selected attendees before, during and after this conference, allowing you to connect to the key people that you want to meet. This online system was designed with your privacy in mind and is only available to registered session attendees of this event.

Hotel & Travel InformationConference Venue:Town and Country Resort & Convention Center500 Hotel Circle NorthSan Diego, CA 92108T: 619-291-7131

Discounted Room Rate: $163 s/d** Room rate includes breakfastDiscounted Room Cut-off: December 22, 2014

Top Reasons to Stay at the Town and Country Resort

• Complimentary wireless internet in your guest room

• Hotel shuttle provided free of charge to hotel guests to locations such as Riverwalk Golf Club, Fashion Valley Mall, Old Town, Mission Valley Center, and Hazard Center (Daily 10am - 7pm, based upon availability)

• Unlimited Bella Tosca Fitness Center access and 10% off Spa & Salon

• Complimentary 3-day trolley pass and overnight parking upon check-in

• Only 10 minutes from San Diego International Airport

Please click here to make your sleeping room accommodations. You will need to identify yourself as a Cambridge Healthtech Institute conference attendee to receive the discounted room rate with the host hotel. Reservations made after the cut-off date or after the group room block has been filled (whichever comes first) will be accepted on a space-and-rate-availability basis. Rooms are limited, so please book early.

Hotel Discount ($100 OFF): Reserve Your Hotel Room at the Town & Country Resort and Save $100 off the Conference Registration… ** You must book your reservation under the PepTalk Room block for a minimum of 5 nights at the Town & Country Resort and Conference Center. Only one discount applicable per Hotel Room.

We understand that you have many choices when making your travel arrangements. Please understand that reserving your room in the CHI room block at the conference hotel allows you to take full advantage of the conference sessions, events and networking opportunities, and ensures that our staff will be available to help should you have any issues with your accommodations.

Car Rental Discounts:

Special discount rentals have been established with Hertz for this conference.

• Click here to make your reservation and use our Hertz Discount Number (CV) 04KL0006

• Or Call Hertz directly at 800-654-3131 and reference our Discount Number (CV) 04KL0006

For more information on San Diego, please visit http://www.sandiego.org/





























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ADDITIONAL REGISTRATION DETAILSEach registration includes all conference sessions, posters and exhibits, food functions, and access to the conference proceedings link.Handicapped Equal Access: In accordance with the ADA, Cambridge Healthtech Institute is pleased to arrange special accommodations for attendees with special needs. All requests for such assistance must be submitted in writing to CHI at least 30 days prior to the start of the meeting.To view our Substitutions/Cancellations Policy, go to www.healthtech.com/regdetailsVideo and or audio recording of any kind is prohibited onsite at all CHI events.

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A series of diverse reports designed to keep life science professionals informed of the salient trends in pharmaceutical technology, business, clinical development, and therapeutic disease markets.For a detailed list of reports, visit InsightPharmaReports.com, or contact Rose LaRaia, [email protected], +1-781-972-5444.

Barnett is a recognized leader in clinical education, training, and reference guides for life science professionals involved in the drug development process. For more information, visit barnettinternational.com.

SHORT COURSE PRICING

Commercial Academic, Government, Hospital-affiliated

Single Short Course $699 $399Two Short Courses $999 $599Sunday, January 18 | 5:00-8:00 PMSC1: Production Challenges for Complex Biologics: ADCs, Bispecifics and Fusion ProteinsSC2: The S-Score System: A Tool for Identifying New Cancer Targets for Antibody-Drug TherapySC3: A Rational Approach to Formulation Development of Biologic TherapeuticsSC4: Genome Editing Using CRISPRSC5: Accelerated Stability Testing of BiologicsSC6: Establishing the Business Case for Single-Use and Continuous Processing

Tuesday, January 20 | 5:00-8:00 PMSC7: Targeting of GPCRs with Monoclonal Antibodies SC8: Affecting Effector Function: Engineering the Fc RegionSC9: Protein Aggregation: Mechanism, Characterization and ConsequencesSC10: Transient Protein Production in Mammalian CellsSC11: Materials in Contact with Biologics: Understanding Risk to Quality and SafetySC12: Protein Purification Strategies: Dealing with Proteins that Are Prone to Aggregate SC13: Continuous Processing of Therapeutic Proteins in Single-Use: Technology and Production Concept

CONFERENCE AND TRAINING SEMINAR PRICING

PREMIUM PACKAGE - BEST VALUE! (Includes access to all conferences and Training Seminars Monday – Friday. Excludes Short Courses)

Early Registration Rate until October 24 $2899 $1399Advance Registration Rate until November 21 $3099 $1629Standard Rate after November 21 and Onsite $3299 $1729STANDARD PACKAGE (Includes access to 2 conferences and/or Training Seminars. Excludes Short Courses)

Early Registration Rate until October 24 $2499 $1199Advance Registration Rate until November 21 $2699 $1299Standard Rate after November 21 and Onsite $2899 $1349

BASIC PACKAGE (Includes access to 1 conference or Training Seminar. Excludes Short Courses)

Early Registration Rate until October 24 $1499 $799Advance Registration Rate until November 21 $1699 $849Standard Rate after November 21 and Onsite $1849 $929

CONFERENCE DISCOUNTS

Poster Submission - Discount ($50 Off): Poster abstracts are due by November 21. Once your registration has been fully processed, we will send an email containing a unique link allowing you to submit your poster abstract. If you do not receive your link within 5 business days, please contact [email protected]. *CHI reserves the right to publish your poster title and abstract in various marketing materials and products.

Antibody Society Members: CHI is pleased to offer all Antibody Society Members a 20% discount to attend. Records must indicate you are a Antibody Society member at time of registration. Please Note - Discounts may not be combined.

Protein Society Members: CHI is pleased to offer all Protein Society Members a 20% discount to attend. Records must indicate you are a Protein Society member at time of registration. Please Note - Discounts may not be combined.

REGISTER 3 - 4th IS FREE: Individuals must register for the same conference or conference combination and submit completed registration form together for discount to apply.

Alumni Discount: Cambridge Healthtech Institute (CHI) appreciates your past participation at our conferences. As a result of the great loyalty you have shown us, we are pleased to extend to you the exclusive opportunity to save an additional 20% off the registration rate.

Group Discounts: Discounts are available for multiple attendees from the same organization. For more information on group rates contact David Cunningham at +1-781-972-5472

If you are unable to attend but would like to purchase the PepTalk CD for $750 (plus shipping), please visit CHI-PepTalk.com. Massachusetts delivery will include sales tax.

How to Register: [email protected] • P: 781.972.5400 or Toll-free in the U.S. 888.999.6288

Please use keycode PTK E when registering

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