the role of analytics in the development of advanced ... · n287 0.4% 0.1% 0.2% 0.6% 1.2% 1.2% n316...
TRANSCRIPT
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1Biogen | Confidential and Proprietary
The Role of Analytics in the Development of Advanced Process Controls (APC)Patrick Swann, Ph.D.September 6, 2016
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2Biogen | Confidential and Proprietary
Specifications are “one part” of an overall Control Strategy
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3Biogen | Confidential and Proprietary
APC can enable an Attribute-Based Control Strategy
Improved use of analytics allows focus on specific quality attributes in-process and use of known levers for process controlAn attribute-targeted control strategy using APC can lead to more efficient and effective controlsMore efficient controls can improve availabilityMore effective controls can improve quality assurance
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4Biogen | Confidential and Proprietary
Product Complexity
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5Biogen | Confidential and Proprietary
Traditional Approach - Charge VariantsIEC
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Minutes6.00 6.10 6.20 6.30 6.40 6.50 6.60 6.70 6.80 6.90 7.00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 7.80 7.90 8.00 8.10 8.20 8.30 8.40 8.50 8.60 8.70 8.80 8.90
Acidic isoforms
Main isoforms
Basic isoforms
Marker
Imaging Capillary IEF of an IgG1 Monoclonal Antibody
cIEF
Slide courtesy of Li Zang, Biogen Analytical Development
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6Biogen | Confidential and Proprietary
Product Characterization - Acidic Fraction Collection
• Free Flow Electrophoresis
IEF based separation, direct correlation of fractions with cIEF peaks
• Fractionation of monoclonal antibody acidic species into 3-4 sub-species
6 http://www.ffeservice.com/index.php/technology.htmlSlide courtesy of Li Zang, Biogen Analytical Development
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7Biogen | Confidential and Proprietary
Abso
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Minutes5.50 5.60 5.70 5.80 5.90 6.00 6.10 6.20 6.30 6.40 6.50 6.60 6.70 6.80 6.90 7.00 7.10 7.20 7.30 7.40 7.50 7.60 7.70 7.80 7.90 8.00 8.10 8.20 8.30 8.40 8.50 8.60 8.70 8.80 8.90 9.00
Acidic Fractions Collected by FFE
DSMain peakAcidic fraction 1Acidic fraction 2Acidic fraction 3Acidic fraction 4
Slide courtesy of Li Zang, Biogen Analytical Development
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8Biogen | Confidential and Proprietary
Asparagine Deamidation
1 2 3 4
N287 0.4% 0.1% 0.2% 0.6% 1.2% 1.2%
N316 2.8% 0.7% 2.7% 6.1% 7.2% 7.6%
N362 0.1% 0.1% 0.1% 0.2% 0.2% 0.2%
N385, N390 4.6% 1.4% 1.8% 8.5% 8.1% 8.5%
Trend
Heavy Chain
Acidic FractionsDS
Main Peak
DeamidationChain
DS
Slide courtesy of Li Zang, Biogen Analytical Development
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9Biogen | Confidential and Proprietary
Isoforms DS Main Peak Acidic Fractions1 2 3 4
Light ChainCML (+ 58 Da) ND ND ND 0.4% 0.7% 1.0%Glycation (+162 Da) 0.6% 1.1% 0.6% 1.1% 0.7% 3.2%N-Glucuronyl (+176 Da) 0.4% ND 0.3% 1.0% 1.0% 0.3%
Heavy Chain
CML (+ 58 Da) 1.2% ND 1.4% 2.9% 3.9% 3.6%+ 79-90 Da 0.8% ND 0.5% 1.5% ND ND+ 111-116 Da 0.6% ND 0.6% ND 1.8% ND+ 121-123 Da ND ND ND 0.7% ND ND+ 132 -137 Da 0.6% ND ND ND 1.1% 0.9%Glycation (+162 Da) 3.1% 1.9% 4.1% 3.1% 3.3% 3.0%N-Glucuronyl (+176 Da) 1.1% ND 1.2% 7.2% 7.2% 7.6%Total impurity 6.4% 3.0% 8.7% 17.9% 19.7% 19.6%
Impurity Level by Reduced Intact Mass
Slide courtesy of Li Zang, Biogen Analytical Development
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Published in: David R. Bush; Li Zang; Arseniy M. Belov; Alexander R. Ivanov; Barry L. Karger; Anal. Chem. 2016, 88, 1138-1146.DOI: 10.1021/acs.analchem.5b03218Copyright © 2015 American Chemical Society
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11Biogen | Confidential and Proprietary
Protein Modifications in Acidic Species
Ref: Du, Y., et al. "Chromatographic analysis of the acidic and basic species of recombinant monoclonal antibodies." MAbs 4(5).
Slide courtesy of Li Zang, Biogen Analytical Development
PTM complexity at individual amino acid level >> separatory capability
An attribute-based control strategy can leverage chemical knowledge about factors that cause formation of these modifications Y = f(x)
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12Biogen | Confidential and Proprietary
The way forward
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13Biogen | Confidential and Proprietary
Biotech Manufacture – High Level Model
Product Quality
Process Input
Proc
ess
Process Control • Robust process design • Advanced process control
Raw Material
RM Control• Screening and/or • Control by Vendor• Learn from MFG experience
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14Biogen | Confidential and Proprietary
APC Key Concepts
Process Analytical TechnologyA system for designing, analyzing, and controlling manufacturing through timely
measurements (i.e., during processing)
of critical quality and performance
attributes of raw and in-process materials
and processes with the goal of ensuring
final product quality.
At-line
In-line
On-line
Slide courtesy of Bassam Nakhle, Biogen Analytical Development
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15Biogen | Confidential and Proprietary
APC Key Concepts
DS DP
Real Time Release Testing (RTRT)The ability to evaluate and ensure the quality of in-process and/or final product based on process data, which typically include a valid combination of measured material attributes and process controls.
Slide courtesy of Bassam Nakhle, Biogen Analytical Development
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16Biogen | Confidential and Proprietary
APC applied to Biotech Manufacture
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17Biogen | Confidential and Proprietary
15kL3750L950L
Seed train
Production Reactor Harvest
Raw Materials
Purification Filling
DS R
elease
DS• Endotoxin• Bioburden• Potency
APC for Biotech Drug Substance
In-line: protein concentrationAt line: purity, aggregation,
charge variants
Raw Materials• Geneology• Spectroscopic tools• Rapid ID testing
Media/buffer Prep control
UF/DF: in-line Protein concentration
Bioreactor: in-line Raman, capacitance
feedback feedback
Feed forward
Bioreactor and/or harvest:At line multi-attribute testing and/or CQA Predictive models for feedforward control
http://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=0ahUKEwjIs9j4sbPKAhXLbT4KHV6XBJUQjRwIBw&url=http://mohali.all.biz/mixing-tank-g343888&psig=AFQjCNF-pMkbyVtS0dboSv-LQTcefVIEig&ust=1453207500280756http://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=0ahUKEwjIs9j4sbPKAhXLbT4KHV6XBJUQjRwIBw&url=http://mohali.all.biz/mixing-tank-g343888&psig=AFQjCNF-pMkbyVtS0dboSv-LQTcefVIEig&ust=1453207500280756
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18Biogen | Confidential and Proprietary
Bioreactor Capture Polish 1 Polish 2 DrugSub.
Drug Prod.
Sialic acid
Deamid-ation
Trisulfidebonds
High mannose
Glycation
AGE
Fragments
Met X oxidation
Etc….
Critical Control Point - a step at which control can be applied and is essential to prevent or eliminate a hazard or reduce it to an acceptable level (ASTM E2590).
Possible Control Points for Charge-Related Attributes
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19Biogen | Confidential and Proprietary
High-Throughput Peptide Map Workflow
30min gradient
Pepfinder, automatic peptide identification
1 2
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Targeted peptide quantitation
Bravo ProA purification (60 min/batch)
Lys-C digestion or rapid trypsin digestion (1-2 hr)
LC –MS analysis, 30 min/sample
Data analysis, 0.5 hr/sample
In-house writtenExcel Script
50-100 µg protein digest
Slide courtesy of Zoran Sosic, Biogen Analytical Development
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20Biogen | Confidential and Proprietary
2-AA analysis (routine)
Peptide Mapping
Flu
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BiN
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Correlation of %Galactosylation with 2-AA
Slide courtesy of Zoran Sosic, Biogen Analytical Development
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21Biogen | Confidential and Proprietary
Controlling Glycation via Glucose
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Low glucose feedback control
High glucose feedback control
Slide – JP Smelko. Data courtesy of Brandon Berry and Terrence Dobrowsky
Glycation = f ([glucose], time, …)
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22Biogen | Confidential and Proprietary
Summary & ConclusionsWhen product complexity is greater than analytical specificity, end-product testing alone is not an attribute-based control strategy
High throughput and selective methods applied to process intermediates can improve process understanding
This process understanding allows a focus on known “levers” (critical control points and critical process parameters) for more efficient and effective controls of product quality attributes
Attribute = f (x1, x2 ,…)
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23Biogen | Confidential and Proprietary
AcknowledgementsRohin Mhatre – Regulatory Affairs
Brian Fahie – Analytical Development• Li Zang• Zoran Sosic• Bassam Nakhle
Eliana D. Clark – Manufacturing Sciences
John Paul Smelko – Cell Culture Development
The Role of Analytics in the Development of Advanced Process Controls (APC)Specifications are “one part” of an overall Control StrategyAPC can enable an Attribute-Based Control StrategyProduct ComplexityTraditional Approach - Charge Variants�IECProduct Characterization - Acidic Fraction CollectionAcidic Fractions Collected by FFE Asparagine DeamidationImpurity Level by Reduced Intact MassSlide Number 10Protein Modifications in Acidic SpeciesThe way forwardBiotech Manufacture – High Level ModelAPC Key ConceptsAPC Key ConceptsAPC applied to Biotech ManufactureSlide Number 17Critical Control Point - a step at which control can be applied and is essential to prevent or eliminate a hazard or reduce it to an acceptable level (ASTM E2590).High-Throughput Peptide Map WorkflowSlide Number 20Controlling Glycation via GlucoseSummary & ConclusionsAcknowledgements