introduction to analytical method development and validation for therapeutic proteins - sample...
DESCRIPTION
Training Seminar: Introduction to Analytical Method Development and Validation for Therapeutic Proteins Wednesday, January 21, 8:30 AM-4:25 PM - Thursday, January 22, 8:30 AM-12:30 PM Part of PepTalk: The Protein Science Week This 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 AD, QC, PD 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 analyses. 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. For full details visit www.chi-peptalk.com/peptalk_content.aspx?id=140090&libID=140061TRANSCRIPT
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Analytical Method Development & Analytical Method Development & Validation for Therapeutic ProteinsValidation for Therapeutic Proteins
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Jichao (Jay) Kang, Ph.D., RAC
Director, Analytical and Formulation Development
Gallus Biopharmaceuticals NJ, LLC
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Course ObjectivesCourse Objectives
Basic scientific and industrial knowledge of working on proteins (including antibodies)
Get familiar with commonly used analytical methods: purpose, advantages, limits, pitfalls
Get familiar with Protein CharacterizationGet familiar with method validation and method
transfer: principle, common practice, and potential problems
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Basic Structure of MAbBasic Structure of MAb
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Major N-linked GlycansMajor N-linked Glycans
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Classified depending on the extent of processing:High mannose: lack Gal or GlcNAc at the end of antennaeComplex: GlcNAc after the Man at the antennaeHybrid: have both Man and Gal/GlcNac at end of the antennae
Hybrid and Complex types can have two or more branches, termed Bi-antennary, tri-antennary, or Tetra-antennary
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Reduced CE-SDS of MAbReduced CE-SDS of MAb
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SEC-HPLC-MALSSEC-HPLC-MALS
Three detector monitoring
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Hydroxylated Proline and Lysine by AAAHydroxylated Proline and Lysine by AAA
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Protein Structure Elucidation by MS/MSProtein Structure Elucidation by MS/MS
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Roepstorff Nomenclature Scheme
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Secondary Structure by CDSecondary Structure by CD
9T Estey, J kang et al., Pharm Sci , 2006, 95: 1626–1639
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Timing for Method Timing for Method Qualification/ValidationQualification/Validation
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Preclinical Clinical Commercial
Phase I Phase II Phase III
Method Development and Qualification
Qualification Validation Verification
Measurement defined
Scientifically Defendable
Ready for validation
Fully ValidatedMethod Transfer
Methods related to product safety require qualification even at phase IOther methods need to be validated prior to BLA, but by phase III, you should
have the methods that can be validated
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Case Study: Binding Assay QualificationCase Study: Binding Assay Qualification
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x axis
1 10 100 1000 10000 100000
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1
2
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Parallel Line Analysis
4-P Fit: y = (A - D)/( 1 + (x/C)^B ) + D: A B C D Rel. Pot.
Standard (Standard: Concentration vs Mean... 0.0395 1.31 202 2.61 1
Sample3 (150%: Concentration vs MeanVal... 0.0395 1.31 127 2.61 1.59
Sample1 (50%: Concentration vs MeanValue) 0.0395 1.31 372 2.61 0.541
Sample2 (100%: Concentration vs MeanVal... 0.0395 1.31 200 2.61 1.01__________
Curve Fit Option - Fixed Weight Value
PLA (Std. Curve: Standard) Degrees of Freedom: parallel = 37 free = 28 non-parallel = 9
R^2 = 1 F-stat = 2.14 F-prob = 0.712
Binding ELISA curves at different level
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Quiz # 15Quiz # 15
For an identification method, what are the validation parameters?
How to determine if a method is stability-indicating?
How to determine LOQ?How to calculate method supported
specification (SS)?List three considerations in setting SSTList three method transfer pitfalls
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