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CASSS WCBP 2012: 16th Symposium on the Interface of Regulatory and Analytical Sciences for Biotechnology Health Products January 23 - 25, 25 2012 Reference Standards for Monoclonal Antibodies: Key Challenges g Addressed Matthew Borer, Ph.D. Advisor Analytical Sciences Research and Development Eli Lilly and Company The Role of Reference Standards: Pharmaceutical Analysis Reference Standards are developed p as p part of the analytical control strategy for each drug product Each reference standard has a control strategy of its own Reference Standards play a central role in assuring the quality of medicines for patients during cGMP testing and release activities WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 2 Typical RS Packaging Options Bulk containers dispense upon customer order Stable compounds No handling issues Less expensive materials Infrequent q use Prepackaged powder Screw-cap bottle Flame sealed ampo Flame-sealed ampoule le Efficient dispensing (resources & materials) Convenient for lab (end-use (end use dating) Protection from oxygen and moisture Content per vial Content-per-vial Shipping “limited quantity exception” Difficult to weigh Hygroscopic, flocculent Lyophilized Expensive or limited supply T i Toxic WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 3 Key Challenge: Packaging Solution So u o p presentation ese a o typically yp ca y mandatory a da o y Poly-cryovials Re-closable Re closable • No special filling equipment • Compatibility testing • – CO2 ingress – adsorption – extractables Cryo-compatible glass ampoules • • • • • • WCBP 2012, 25-Jan-2012, M. Borer, et al. Hermetic seal Inert atmosphere F Fewer issues i with ih adsorption/extractables Less expensive Single use Special filling equipment © 2012 Eli Lilly and Company 4 Key Challenge: Sterility Is Sterility Required? The reference Th f standard t d d is i used d iin cell-based ll b d assays where h contaminant t i t organisms can proliferate • Contaminant organisms can grow and degrade peptides on storage • Or Not? Reference materials are not drug products, so patient safety is not impacted • Aseptic manufacturing is difficult and facilities might not accept some source materials • An Answer: Sterile filter the bulk reference material source solution • Prepare the reference materials in a clean environment • Store reference materials frozen • Include a broad-spectrum antibiotic in the cell culture medium • WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 5 Key Challenge: Source Material Selection Development batch or batch used in toxicological testing. May be sourced d ffrom mixed pool of clones. First Laboratory RS Recommended to replace the initial batch with one derived from the FHD batch, or a batch that is representative. New Batch First Human Dose ((FHD)) WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company Replace batch only when projected to stock-out or in the eventt off a significant i ifi t process change. New Batch Commercial Development First Efficacy y Dose ((FED)) 6 Source Material: Commercialization Derived from, or representative of a batch used in pivotal clinical of, studies. Represents the final optimized commercial process. Compendial p RS New Batch Primary RS ((unlikely y event)) Secondary RS New Batch Formal Stability Prefer to source from same packaging k i run as the first Primary RS. WCBP 2012, 25-Jan-2012, M. Borer, et al. New Batch Global Registration Subsequent Secondary RS sourced d ffrom a representative batch. © 2012 Eli Lilly and Company 7 Key Challenge: Characterization Reference standard characterization must be customized to support its specific intended use ICH Q3a, IV • Reference R f standards t d d used d iin th the analytical l ti l procedures d ffor control t l off iimpurities iti should h ld be evaluated and characterized according to their intended uses. uses WHO, Annex 3, Technical Report Number 885,3 • It is necessary to consider all data obtained from testing the material by a wide variety of analytical methods. When taken as a whole, this will ensure that the substance is suitable for its intended use. use The extent of the analyses required depends on the purpose(s) for which the chemical reference substance is to be employed, and may involve a number of independent laboratories. ISO 34, 4.1.1 • It should be recognized that a reference material needs to be characterized mainly to the level of accuracy required for its intended purpose WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 8 Typical mAb Characterization Tests Appearance pp Purityy Identity • Light g and Heavy y Chain MS Protein Quantity • Oligosaccharide Profile UV • Charge Heterogeneity • Size-exclusion HPLC • Reduced SDS-PAGE (or CESDS)) • Capillary Electrophoresis-LIF Potency Ion-exchange g chromatography g p y • Bioassay pH WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 9 The Characterization Challenge No wayy to fullyy define the Potencyy of a mAb using g physiochemical testing, so the Primary RS defines biological activity but The Primaryy RS has no basis for comparison, p , so monitoring for change in Potency is hampered and Bioassay methods are typically highly variable, making it difficult to measure small changes WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 10 Potency Assignment Strategy: Initial Batch Laboratory RS First Laboratoryy RS is assigned g a value of 100% Relative Potency (RP) • Primary RS Subsequent Laboratory RS batches are assigned relative to the previous Laboratory RS The Primary RS is assigned a value of 100% RP Confirmation of a dose response relationship • For-information-only Bioassay comparison to Laboratory RS • – more extensive for IgG1 mAbs due to greater potential for potency change • Secondary RS Extensive physicochemical comparability testing to Laboratory RS The initial Secondary RS is based on the same characterization tests as the Primary RS (same source material) • A subsequent Secondary RS is assigned a value of 100% RP if the bioassay versus the Primary RS is within a pre-defined window WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 11 Potency Assignment Strategy: Replacement Batch Laboratory RS Analyzed y by y the biological g assay y using g 5 independent p measurements relative to the previous RS If the average relative potency is within 80-120% 80 120% RP RP, the new reference standard will be assigned a value of 100% RP If the average potency falls outside of the interval, an investigation is conducted. Question the suitability of the material • Consider a larger number of replicate measurements in order to assign the RP directly to the previous standard • WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 12 Potency Assignment Strategy: Replacement Batch Secondary RS Analyzed y by y the biological g assay y using g 10 independent p measurements relative to the Primary RS If the average relative potency is within 95-105% 95 105% RP RP, the new reference standard will be assigned a value of 100% RP If the average potency falls outside of the interval, an investigation is conducted Question the suitability of the material • Consider a larger number of replicate measurements in order to assign the RP directly to the previous standard • WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 13 Independent Measurement Defined Independent p Measurement - In General • An average reported result based on a replication strategy that encompasses appropriate variance components associated with the measurement Independent Measurement for Bioassay Three plates • Three replicates at each dose level for each: • Sample • Standard • Control C sample • WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 14 Key Challenge: Ongoing Suitability for Use (a.k.a. (a k a stability) Comprehensive physicochemical testing Monitoring bioassay curve parameters over time Overprotective packaging and storage temperature Assess representative stability (e.g., from the commercial process) Stress stability Promote the implementation of higher order standard (e.g., WHO International Standard) Careful control of bioassay • • • • Use of an independent control material Control over key reagents Consistent technique and comprehensive analyst training Control over instrumentation WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 15 Future Challenges The difficulties associated with characterization of biomolecule reference standards make harmonization of multiple compendial reference standards a challenge Global manufacturers must meet regional standards • Different doses of the same drug in different regions does not make sense for patients • Differences in compendial RS assignments can happen • – Variability in measurements – Differences in the characterization approach This will be a particular challenge for monoclonal antibodies WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 16 Co Authors Co-Authors Kimberley y B. Dancheck Kristi L. Griffiths Bryan J. Harmon Jerry J. Lewis Jerry Zhirui Lian Bhavin S. Parekh WCBP 2012, 25-Jan-2012, M. Borer, et al. © 2012 Eli Lilly and Company 17