Product and Process Variants & Impurities

Wednesday, October 23, 2013 • Plenary Session8:30 Chairwoman’s RemarksNadine M. Ritter, Ph.D., Senior CMC Consultant, Biologics Consulting GroupHost Cell Protein and Host Cell DNA Characterizationand Control8:15 FDA Perspective Regulatory Perspectives onHost Cell Protein Characterization and ControlLaurie Graham, Acting Team Leader, Product Quality/CMC, Division ofMonoclonal Antibodies, CDER US FDA8:45 UNPUBLISHED DATA New USP Chapters for Measurementof Residual Host Cell Protein and DNA Impurities inBiotherapeuticsUSP will soon propose a new test chapter Residual DNA Testingcontaining a validated method with two new Reference Standards for CHOand E. coli genomic DNA. Another new chapter contains best practices fordevelopment, validation, and use of residual HCP testing methods. Detailsof the chapters are presented.Maura C. Kibbey, Ph.D., Senior Scientific Liaison, Biologics &Biotechnology, US Pharmacopeia9:15 Technology WorkshopThe Devil in the Detail –HCP Assay Performance by DesignAlthough immunoassays for HCP determination are well established in QCapplications, there are some inevitable risk factors (antigen, animals etc.)associated with this approach. This presentation will highlight challenging casestudies from more than twenty years of experience in HCP assay development.Critical parameters and quality attributes along the assay development processwill be examined along with newly emerging technologies.Michael Hantman, Ph.D., Associate Director Methods Development,Charles River9:45 Impact of Host Cell Proteins on GMP Testing and ProcessCharacterization/Process ValidationAs the pace quickens to move molecules through the pipeline to market, thequality and integrity of drug products must be maintained. The accuratequantification and identification of residual host cell proteins in GMP andPC/PV materials provide multiple challenges to today’s testing paradigm.Lori O’Connell, Manager, Analytical Operations, Genentech, Inc.,A member of the Roche Group10:15 Technology WorkshopUse of QPCR and DNA Sequencing Toolsto Ensure Product Quality and SafetyQuantitative PCR (Q-PCR) and DNA sequencing are tools that enable rapid,sensitive and precise quantitation, detection and identification of critical cellularand process impurities in cell culture manufacturing and product purification.Additionally, these tools can be utilized in development and characterizationof production cell lines and in routine monitoring of cell line stability. Inthis presentation, the applications of these technologies and present datademonstrating the performance of assays for impurity assessment, cell linecharacterization, contaminant detection and identification are reviewed.Wesley Straub, Ph.D., Senior Product Manager, Bioproduction,Life Technologies10:45 Networking Refreshment BreakCharacterization of Degradation Pathways11:15 Utilization of Forced Degradation Data in ProductRegulatory FilingsNadine M. Ritter, Ph.D., Senior CMC Consultant, Biologics Consulting Group11:45 CASE STUDY Predict Protein Thermal Aggregation Kinetics byDifferential Scanning FluorimetryDifferential scanning fluorimetry (DSF) is an emerging technique to studyprotein thermal unfolding and has a great potential to be employed as botha protein characterization tool and a first-pass high-throughput screening(HTS) assay in formulation screening and development. In a case study,we demonstrate how DSF together with analytical ultracentrifugation(AUC) can predict protein concentration dependent thermal aggregationkinetics. In another case study, we show the importance of establishingan appropriate correlation upfront in understanding different aggregationbehaviors at high concentration for different proteins.Shuai “Sunny” Shi, Ph.D., Senior Scientist, Sterile Product Development, Merck12:15 CASE STUDY/UNPUBLISHED DATA Assessment of Potential ProteinDegradation Sites in the CDR of Recombinant AntibodiesDegradation of proteins by asparagine (Asn) deamidation and methionine(Met) oxidation can impact in vivo biological functions and in vitro stability oftherapeutic antibodies (mAbs). In the present study, an approach employingstress conditions (elevated temperatures, pH, and oxidizing agents) andproteolytic peptide mapping combined with quantitative LC-MS for theinduction, identification and quantification of Asn deamidation and Metoxidation was developed. This test system allowed us to identify a light chainAsn and heavy chain Met, both located in the CDR 3, as potential chemicaldegradation sites in a recombinant IgG1.Markus Haberger, Manager Development Characterization,Roche Diagnostics GmbH, Germany12:45 Lunch on your ownBiophysical and Structural Characterization Strategies2:00 FDA Perspective Particulate Testing and Specifications forSubvisible and Visible ParticulatesParticulate matter in protein therapeutics may originate from various sourcesand it may affect the safety and efficacy of the products. Limiting the amount ofparticulates and ensuring product consistency through adequate testing reducesthe risks related to their presence. Recent advances in understanding the safetyof particulate matter with focus on protein aggregates, testing methodologiesand trends in setting standards will be discussed.Ewa Marszal, Ph.D., Chemist, Laboratory of Plasma Derivatives, Division ofHematology, CBER, US FDA2:30 Development of Compendial Analytical Procedures forN-Glycan AnalysisThe United States Pharmacopeia is developing a new General Chapter to providequalitative analysis of glycosylation through profiling of released N-linkedoligosaccharides (or N-glycans). This chapter consists of validated analyticalprocedures and performance criteria. Furthermore, four reference standards havebeen proposed to assess the system suitability for the analytical procedures.Edith Chang, Ph.D., Reference Standards Scientist, Biologics andBiotechnology, US Pharmacopeia3:00 NMAP: A Novel Approach to Understanding MolecularStructure Using Native Peptide MappingSpectroscopic techniques such as circular dichroism and intrinsic fluorescence arecommonly used to assess the higher order structure analysis of proteins. However,detailed molecular understanding is challenging as these techniques provide structuralensemble data. The use of orthogonal methods is essential to understand complexbiological molecules. An orthogonal method, Native Peptide Mapping (NMap),was developed on the basis of a traditional peptide mapping in order to addresslocal structural understanding. The evaluation of the potential of NMap to generatetertiary structure related data, as well the correlation of the results with traditionalspectroscopic techniques using a drug substance monoclonal antibody is discussedin this presentation. The proposed use of NMap would include characterizationunderstanding and application for comparability data and submissions.John O’Hara, Ph.D., Director, Characterization and Method Development,Analytical Sciences, Biologicals, UCB, United Kingdom3:30 Panel Discussion:Sensitivity of Methods to Detect Variants and Impurities• Why is accurate validation and system suitability of LOQ critical for variantand impurity assays?• How do you bridge old methods to new methods for variants and impurities?• What should you do if more sensitive methods cause increased detection ofsmall amounts of impurities/variants?• How does a biosimilar product assess comparability to the variants andimpurities of the innovator product?Moderator:Nadine M. Ritter, Ph.D., Senior CMC Consultant, Biologics Consulting GroupPanelists:Ewa Marszal, Ph.D., Chemist, Laboratory of Plasma Derivatives,Division of Hematology, CBER, US FDAMikhail V. Ovanesov, Ph.D., Visiting Scientist, Principal Investigator, Laboratory ofHemostasis / Division of Hematology / Office of Blood Research and Review,CBER, US FDAAnton V. Manuilov, Ph.D., Sr. Scientist II, Protein Analytics,AbbVie Bioresearch CenterJudy Shimoni, Ph.D., Senior Scientist, PTDU - Protein Analytical Chemistry,Genentech, Inc.Kirk J. Leister, Ph.D., Director of New Technology, Bristol Myers Squibb Company4:00 Close of Conference10 www.IBCLifeSciences.com/WCB www.IBCLifeSciences.com/Variants