PIONEERING CELL & GENE THERAPY SOLUTIONSFROM DISCOVERY TO THE CLINIC

Enrichment of NK Cells from PBMCs with the MagCellect Human NK Cell Isolation Kit (Catalog # MAGH109).

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CELL SEPARATION AND CHARACTERIZATION

CELL CULTUREGMP ancillary and raw materials must provide robust performance in cell manufacturing workflows. With our focus on quality, innovation, supply chain continuity, and traceability, Bio-Techne® GMP-grade reagents are the reliable solution for your cell culture expansion, activation, and differentiation protocols (see pages 5 and 6).

Before Enrichment

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CUSTOM SOLUTIONSEfficient workflows often require customized solutions. Put our decades of experience to work for you. We offer GMP conversion of common antibodies used for cell separation plus many other services for cell therapy ancillary and raw materials.

CUSTOM SOLUTIONSLearn about our services for cell therapy ancillary and raw materials, including custom GMP protein bottling, ligand conjugation to Cloudz™ dissolvable microspheres, and GMP conversion for your specialized media and supplements.

Obtaining and qualifying a robust starting cell population is the essential first step for an ex vivo manufacturing workflow. By coupling innovative cell selection technologies with premium flow cytometry antibodies for cell qualification, Bio-Techne enables high-purity enrichment of immune cell and stem cell subsets.

• Multi-color flow cytometry kits for identification of cell lineages and subsets

• Extensive catalog of immune cell characterization antibodies for flow cytometry

• GMP proteins including ProDots® formulations

• GMP small molecules

• GMP media

• GMP Cloudz™ T Cell Activation Kit

• Excellerate™ Human T Cell Expansion Media, Xeno-Free – GMP coming soon!

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Gene Transposition in Primary T Human T Cells. Primary human T cells were collected from 4 donors and a Chimeric Antigen Receptor (CAR A, CAR B) or T Cell Receptor (TCR) was transposed into the donor cells using a non-viral transposon-based system from R&D Systems. Data show average integration rates of 30% across the donors or cargo type.

Correlation Between Immunoassay Platforms for Smooth Transition to Automated Analyte Testing. High-throughput Simple Plex™ assays are calibrated to Quantikine® ELISA kits during development.

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GENE EDITINGGenetic engineering of pluripotent stem cells and designing CAR-T cells for immunotherapy are key steps in developing cell therapies. Our non-viral TcBuster™ transposon technology enables stable integration of genetic constructs (see page 7).

PRODUCT RELEASE TESTINGA safe and consistent cell therapy product requires stringent, sensitive, and standardized assays to document purity and quality. Our expertise and pioneering technologies allow you to generate reliable and accurate data for release testing while ensuring your cell therapy gets to the clinic as fast and as safely as possible.

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CUSTOM SOLUTIONSLearn about our gene editing services including HLA deletion for immune masking and flexible cell validation methods. We offer dedicated project management supported by the Custom team at R&D Systems.

TcBuster Efficiency in Delivering CARs and TCRs into Primary Human T Cells

GENE EDITING CAPABILITIES WITH TCBUSTER

• Fastest turnaround in the industry

• Under 5 months to prepare for GMP manufacturing

• High frequency, high efficiency editing

• Knock-in, knock-out, and KI/KO combinations

• No cell line is too difficult for us to engineer

• Customizable—HLA, gender, or CMV status of donor cells are available upon request

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Confirmation of antigen specificity and surface expression of anti-CD19 CAR construct.

Analysis of particle morphologies in a manufacturing sample.

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Confirm that the cells in your product exhibit the correct phenotype with flow cytometry or automated immunoassays. Verify CAR expression plus marker proteins and secreted analytes.

PRODUCT PURITY TESTINGMake sure your final product meets purity specifications by quantitating and characterizing particulate and droplet contaminants.

Micro-Flow Imaging™ provides image-based analysis of subvisible particle counts and morphology. These instruments discriminate between protein aggregates, oil droplets, bubbles, and other contaminants.

• Up to 150 µL/minute at 900,000 particles/mL

• Bot1 Autosampler for up to 90 samples per run

• High-resolution images with 85% sampling efficiency

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IMMUNOASSAYS FOR CELL PHENOTYPE TESTING

Simple Plex™ automated ELISAs are ideal for in-process and release testing for cell therapy products. Ella provides unparalleled accuracy, reproducibility, and speed for characterizing cytokine secretion profiles.

Direct-labeled Fluorokine™ ligands bind to chimeric antigen receptors to verify the antigen specificity of your CAR. We also offer thousands of conjugated primary antibodies and custom antibody conjugation with nearly 20 flurophores.

• Automated quantitation of cytokine secretion profiles in <1 hour

• 4-5 log dynamic range and sub-picogram detection sensitivity

• Samples run in individual channels to eliminate cross-reactivity

• Customizable multiplex cartridge options

• 21 CFR Part 11-compliant

• Flow cytometry to confirm integrity of CAR

• Most common CAR binding specificities

• Additional Fluorokine™ ligands available by request

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GMP ANCILLARY REAGENTSTraceability, scalability, and bioactivity are essential for cell manufacturing supplements. GMP ancillary reagents from Bio-Techne are designed with all three in mind.

GMP Small Molecules - We are the premium source of GMP small molecules! Let us help you develop the next stem cell therapy.

• Y-27632 (ROCK inhibitor) - enhancing stem cell survival and viability

• SB 431542 (TGF-b RI inhibitor) - differentiation and reprogramming protocols

• CHIR 99021 (GSK-3 inhibitor) - differentiation and reprogramming protocols

• DAPT (γ-Secretase inhibitor) - neuronal differentiation protocols

• XAV 939 (Tankyrase inhibitor) - cardiomyocyte differentiation protocols

GMP Cell Culture Media - Our GMP media is manufactured so that all products, materials, equipment, and processes used in production are traceable and controlled.

• GMP ExCellerate™ Human T Cell Expansion Media, Xeno-Free – coming soon

• StemXVivoTM GMP Mesenchymal Cell Media

• GMP N-2 MAX Media Supplement for culturing neurons, neural progenitors, and stem cells

CERTIFICATION AND REGULATORY GUIDELINES FOLLOWED FOR GMP REAGENTS

• Proteins are manufactured in our ISO 9001:2015, ISO 13485:2016-certified facility.

• Small molecules are manufactured following a quality system based on ICH Q7 guidelines.

• USP Chapter <1043>, Ancillary Materials for Cell, Gene, and Tissue-Engineered Products

• USP Chapter <92>, Growth Factors and Cytokines Used in Cell Therapy Manufacturing

• Ph. Eur. General Chapter 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products

GMP Recombinant Proteins - We often use the same clone, sequence, and expression system as our research grade materials. This makes the switch to GMP as seamless as possible.

• IL-2, -3, -4, -6, -7, -12, -15

• Activin A, FGF basic, Noggin, SCF

• And many more … an industry-leading selection of nearly 50 GMP proteins!

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PIONEERING TECHNOLOGIES FOR CELL CULTURE

GMP CLOUDZ™ CELL ACTIVATION AND EXPANSION KITSCloudz microspheres are a novel cell activation technology that provide safety and efficacy advantages over current bead-based cell activation methods.

GMP-GRADE PRODOTS® PROTEINSThese pre-aliquoted, lyophilized, and instantly dissolvable spheres of animal-free GMP cytokines or growth factors are closed-system ready.

• Eliminates protein aliquoting

• Reduces unused reconstituted protein waste

• Quickly welds into standard closed-system tubing

• Streamlines protocols and limits handling errors

• Improves efficiency of GMP protein supplementation

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• GMP Cloudz™ Human T Cell Activation Kit

• Human Treg Expansion Kit

• Human NK Cell Expansion KiT

Equal Performance Activity of GMP ProDots Compared to Standard GMP Proteins. Human T cells were expanded for 9 days from peripheral blood mononuclear cells using either GMP ProDots IL-2 or standard GMP IL-2 in combination with ExCellerate™ T Cell Media and the Cloudz™ T Cell Activation Kit.

Comparison of Cloudz Human T Cell Activation Kit with immobilized CD3 and CD28 antibodies. Stimulation of IFN-gamma secretion by CD4+ T cells.

• Dissolvable microspheres conjugated with cell-activating antibodies or ligands

• Robust cell activation and expansion

• Release buffer dissolves particles in < 10 seconds

• Simplified recovery of cells – no beads to remove from cell culture!

• Customizable with other antibodies and proteins for your process

• Compatible with multiple clinical expansion platforms

GMP PRODOTS PROTEINS FEATURES

• Each bag contains a pre-aliquoted amount of GMP ProDots Protein

• Bags contain weldable tubing for sterile docking and a syringe port

• GMP ProDots dissolve instantly when cell culture media or buffer is added to bag

• Manufactured under animal-free and GMP controlled conditions

• Aseptically packaged into single-use bags and tested to USP <71> guidelines

• GMP ProDots coming soon: IL-2, IL-7, IL-15, IL-21

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PIONEERING TECHNOLOGIES FOR GENE EDITING

TCBUSTER™ NON-VIRAL GENE EDITINGThis novel and flexible transposon-based gene delivery system is designed to simplify and improve the efficiency of gene editing for therapeutic manufacturing. TcBuster offers a number of benefits over viral editing methods, including:

Electroporation of TcBuster platform

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T cells were transposed using TcBuster and subsequently expanded. Cells were transposed using a single transposon encoding multiple genes demonstrating the ability of TcBuster to deliver large cargo. In A) the transposon encoded a TCR and 2 different accessory proteins and in B) the transposon encoded 2 independent CARs.

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Schematic overview of TcBuster mechanism of transposition. 1. TcB-E transposase and transposon are introduced into the cell. 2. Protein from TcB-E mRNA is produced. TcB-E transposase cuts the cargo from the transposon plasmid. 4. TcB-E transposase pastes the transposon cargo into the genomic DNA. 5. Cargo mRNA is stably expressed from the genomic DNA. This example illustrates stable expression of a receptor protein, such as a CAR.

• Accommodates larger cargo enabling multi-gene delivery

• Lowers cost per patient

• Ensures steady and scalable supply

• Improves post-editing cell viability and expansion

• Transposons have demonstrated the safest insertion profile of any gene delivery system

• Available to deliver Your Favorite Genes… in Your Favorite Cell™

TcBuster overcomes the bottleneck of global GMP capacity associated with custom virus-based manufacturing.

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WHERE SCIENCE INTERSECTS INNOVATION™

bio-techne.com

Global info@bio-techne.com bio-techne.com/find-us/distributors TEL +1 612 379 2956 North America TEL 800 343 7475 Europe | Middle East | Africa TEL +44 (0)1235 529449 China info.cn@bio-techne.com TEL +86 (21) 52380373

For research use or manufacturing purposes only. Trademarks and registered trademarks are the property of their respective owners.

BR_CGT Workflow_CGT1010

Your vision is to create revolutionary cell and gene therapies to treat life threatening diseases. Bio-Techne and its family of brands is on this journey with you. As a full-solution ancillary reagent and services provider, we will stand by you, providing flexible and pioneering tools to simplify your workflow. From CAR T cells to pluripotent stem cells, let us help you get your therapy to the

patients that need it!