Well Characterized Biologicals
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October 20 - 22, 2010 · Hyatt Regency Bethesda · Bethesda, MD
Shared Exhibit Hall with Process2Product: Translating Process Knowledge into Quality Biological Products
Document Title
Agenda
7:00
Registration and Coffee
8:00
Chairperson's Welcome and Opening Remarks
Nadine M. Ritter, Ph.D., Senior CMC Consultant, Biologics Consulting Group, Inc.
Nadine M. Ritter, Ph.D., Senior CMC Consultant, Biologics Consulting Group, Inc.
Characterizing Complex Protein Products
8:15
Designing an Effective Comparability Study: A Case Study for a Novel Protein Therapeutic
Products with an extended development history can face significant challenges when approaching registration, particularly for small pharmaceutical companies. For many products, cutting edge analytical technology is not always available during early phase development, so bridging the early to late phase process through parallel testing is often necessary. In some cases, the fear of what may be found can deter a company from providing an adequate comparability package, thus jeopardizing the quality of the submission and the likelihood of approval. However, as the truth can set you free, so can a well designed and executed comparability package. This case study will describe the design and execution of a successful comparability study for a recently approved, novel and complex protein therapeutic.
Asenath M. Rasmussen, Ph.D., Associate Research Fellow, Analytical Research & Development, Pfizer, Inc.
Products with an extended development history can face significant challenges when approaching registration, particularly for small pharmaceutical companies. For many products, cutting edge analytical technology is not always available during early phase development, so bridging the early to late phase process through parallel testing is often necessary. In some cases, the fear of what may be found can deter a company from providing an adequate comparability package, thus jeopardizing the quality of the submission and the likelihood of approval. However, as the truth can set you free, so can a well designed and executed comparability package. This case study will describe the design and execution of a successful comparability study for a recently approved, novel and complex protein therapeutic.
Asenath M. Rasmussen, Ph.D., Associate Research Fellow, Analytical Research & Development, Pfizer, Inc.
8:45
Strategies in the Design of Potency Assays for Therapeutic Enzymes
Potency assays for therapeutic enzymes should be highly relevant to the proposed mechanism-of-action. Ideally, good potency assays for therapeutic enzymes are indicators of enzyme structure, enzymatic activity and appropriate cellular targeting. This presentation will discuss strategies of potency measurements for therapeutic enzymes.
Richard Ledwidge, Ph.D., Drug Reviewer, Division of Therapeutic Proteins, CDER, US FDA
Potency assays for therapeutic enzymes should be highly relevant to the proposed mechanism-of-action. Ideally, good potency assays for therapeutic enzymes are indicators of enzyme structure, enzymatic activity and appropriate cellular targeting. This presentation will discuss strategies of potency measurements for therapeutic enzymes.
Richard Ledwidge, Ph.D., Drug Reviewer, Division of Therapeutic Proteins, CDER, US FDA
New, Unpublished Data
9:15
Characterization of Calcium-binding as a Useful Signature of Structure-function for a Factor IX Fc-fusion Protein
Calcium-binding to Factor IX imparts structural properties essential to its role in the blood clotting cascade. Factor IXFc is a heterodimeric fusion protein designed for a prolonged therapeutic half-life. Biophysical studies of structural changes that accompany calcium-binding will be presented and considerations for product characterization and comparability will be discussed.
Mark Brader, Ph.D., Principal Scientist, Protein Pharmaceutical Development, Biogen Idec
Calcium-binding to Factor IX imparts structural properties essential to its role in the blood clotting cascade. Factor IXFc is a heterodimeric fusion protein designed for a prolonged therapeutic half-life. Biophysical studies of structural changes that accompany calcium-binding will be presented and considerations for product characterization and comparability will be discussed.
Mark Brader, Ph.D., Principal Scientist, Protein Pharmaceutical Development, Biogen Idec
New, Unpublished Data
9:45
Analytical Resolution/Characterization of Conjugated Proteins
Using complementary methods can be an effective means toward the analytical characterization of proteins. Conjugated proteins add challenges, depending on the conjugation chemistry, site(s), and conjugating molecule(s). Conjugations at multiple sites add additional complexity, however site-specific conjugation through the introduction of novel amino acids as discrete conjugation sites can reduce this complexity. This approach can simplify analysis, allowing evaluation of site-specific effects on pharmacology, efficacy, and biophysical characteristics. Applications will be discussed.
Anna-Maria Hays Putnam, Ph.D., Group Leader, Analytical and Formulation Development, Ambrx, Inc.
Using complementary methods can be an effective means toward the analytical characterization of proteins. Conjugated proteins add challenges, depending on the conjugation chemistry, site(s), and conjugating molecule(s). Conjugations at multiple sites add additional complexity, however site-specific conjugation through the introduction of novel amino acids as discrete conjugation sites can reduce this complexity. This approach can simplify analysis, allowing evaluation of site-specific effects on pharmacology, efficacy, and biophysical characteristics. Applications will be discussed.
Anna-Maria Hays Putnam, Ph.D., Group Leader, Analytical and Formulation Development, Ambrx, Inc.
10:15
Networking Refreshment Break
10:45
Characterization of a Complex Protein Product
Abstract not available at time of print.
Jihong Wang, Ph.D., Senior Scientist, Analytical Biochemistry, MedImmune, Inc.
Abstract not available at time of print.
Jihong Wang, Ph.D., Senior Scientist, Analytical Biochemistry, MedImmune, Inc.
New, Unpublished Data
11:15
Characterization of a Novel Combination of Two Anti EGFR Antibodies Showing Superior Anticancer Efficacy
We have developed Sym004, a product consisting of two anti-EGFR monoclonal antibodies which inhibits cancer cells by blocking ligand binding, receptor activation/phosphorylation, and downstream signaling. In contrast to other mAbs targeting EGFR, Sym004 induces rapid and efficient removal of the EGFR from the cancer cell surface by inducing EGFR internalization and degradation. This presentation will show examples of the release and characterization assays used for analysis of the individual antibodies in Sym004. In addition, a manufacturing strategy for Sym004 will be presented together with examples of release and characterization assays for the combination of two antibodies.
Torben P. Frandsen, Ph.D., Director, Antibody Chemistry, Symphogen A/S, Denmark
We have developed Sym004, a product consisting of two anti-EGFR monoclonal antibodies which inhibits cancer cells by blocking ligand binding, receptor activation/phosphorylation, and downstream signaling. In contrast to other mAbs targeting EGFR, Sym004 induces rapid and efficient removal of the EGFR from the cancer cell surface by inducing EGFR internalization and degradation. This presentation will show examples of the release and characterization assays used for analysis of the individual antibodies in Sym004. In addition, a manufacturing strategy for Sym004 will be presented together with examples of release and characterization assays for the combination of two antibodies.
Torben P. Frandsen, Ph.D., Director, Antibody Chemistry, Symphogen A/S, Denmark
11:45
Combination Products: An Overview
What are Combination Products and how are they regulated by the FDA? What is the Office of Combination Products and what do they do? Which Center takes the review lead? What are some of the CMC challenges? This talk will attempt to answer all of these questions and provide an introduction to the complex world of Combination Products.
Kathy Lee, Associate Laboratory Chief, Laboratory of Biochemistry, Division of Therapeutic Proteins, CDER, US FDA
What are Combination Products and how are they regulated by the FDA? What is the Office of Combination Products and what do they do? Which Center takes the review lead? What are some of the CMC challenges? This talk will attempt to answer all of these questions and provide an introduction to the complex world of Combination Products.
Kathy Lee, Associate Laboratory Chief, Laboratory of Biochemistry, Division of Therapeutic Proteins, CDER, US FDA
12:15
Q&A Panel Discussion with Morning Speakers
12:45
Networking Luncheon
Structure/Function and Critical Quality Attributes
2:00
Characterization of Biotechnology Products: An FDA Perspective
As described in ICH Q6B, the characterization of biotechnology products should include a determination of physiochemical properties, biological activity, immunochemical properties, purity, and impurities. During product development, characterization studies help to determine those attributes which impact the safety and efficacy of the product. Information on these critical quality attributes, and how they are influenced by the manufacturing process, can then be used establish the product's control strategy and design space. In addition, data from extended characterization studies are typically used to support comparability after significant manufacturing changes. The development of well-developed characterization strategies, which utilize current technologies and assess all relevant, including emerging, quality attributes can, however, be challenging. Ms. Graham will discuss, through various case studies, the current regulatory expectations with regard to characterization strategies for monoclonal antibody products.
Laurie Graham, Product Quality/CMC Reviewer, Division of Monoclonal Antibodies, Office of Biotechnology Products, CDER, US FDA
As described in ICH Q6B, the characterization of biotechnology products should include a determination of physiochemical properties, biological activity, immunochemical properties, purity, and impurities. During product development, characterization studies help to determine those attributes which impact the safety and efficacy of the product. Information on these critical quality attributes, and how they are influenced by the manufacturing process, can then be used establish the product's control strategy and design space. In addition, data from extended characterization studies are typically used to support comparability after significant manufacturing changes. The development of well-developed characterization strategies, which utilize current technologies and assess all relevant, including emerging, quality attributes can, however, be challenging. Ms. Graham will discuss, through various case studies, the current regulatory expectations with regard to characterization strategies for monoclonal antibody products.
Laurie Graham, Product Quality/CMC Reviewer, Division of Monoclonal Antibodies, Office of Biotechnology Products, CDER, US FDA
New, Unpublished Data
2:30
Case
Study
In vitro Biological Characterization of Antibodies with N-linked High Mannose GlycansStudy
In the present study, we evaluated the effects of high mannose glycans on effector functions of an anti-CD20 antibody. Results of our study demonstrate that antibodies with high mannose glycans have enhanced potency in FcγRIIIa binding and ADCC activity but reduced potency in FcγRIIa/IIb binding and CDC activity.
Shan Chung, Ph.D., Scientist, Bioanalytical Research and Development, Genentech, Inc.
3:00
Assessing the Criticality of Antibody Charge-based Variants Using in vivo Monitoring
Abstract not available at time of print.
Gregory Flynn, Ph.D., Scientific Director, Process and Product Development, Amgen, Inc.
Abstract not available at time of print.
Gregory Flynn, Ph.D., Scientific Director, Process and Product Development, Amgen, Inc.
3:30
Networking Refreshment Break
4:00
Case
Study
The Role of Mass Spectrometry in Determining the Potential Critical Quality Attributes (CQA) of a Monoclonal AntibodyStudy
This presentation will discuss two case studies. We used mass spectrometry for later stage research/early development to characterize the mAb CDR related quality attributes and determine the potential CQA status. Later, we developed the lot release assays for those identified CQAs with the assistance of mass spectrometry.
Wenjun (David) Mo, Ph.D., Senior Scientist, Analytical Biochemistry, MedImmune, Inc.
4:30
A Cell-Based FcRn Binding Assay to Assess Fc Functionality of IgG-Based Therapeutics
Abstract not available at time of print.
Abhishek Mathur, Ph.D., Scientist, Bioassay and Biological Characterization, Amgen, Inc.
Abstract not available at time of print.
Abhishek Mathur, Ph.D., Scientist, Bioassay and Biological Characterization, Amgen, Inc.
5:00
Q&A Panel Discussion with Afternoon Speakers
5:30
Close of Day One
8:00
Coffee
8:30
Chairperson's Remarks
Ned Mozier, Ph.D., Senior Director, Analytical Sciences, Global Biologics Research and Development, Pfizer, Inc.
Ned Mozier, Ph.D., Senior Director, Analytical Sciences, Global Biologics Research and Development, Pfizer, Inc.
Biological Assays for Product Characterization
8:45
A Regulatory Perspective on the Development and Validation of Biopharmaceutical Potency Assays
Biologics license applications must demonstrate that the product is safe, pure and potent according to the Public Health Service (PHS) Act 42, USC 262, and the Code of Federal Regulations (21 CFR), 601.2. It is emphasized in the PHS Act under Subpart 1-Biological Products, and 21 CFR 610.10-Potency that all biological drug products are required to be released using an assay that measures their potency. Development of a potency assay that is both reflective of the biopharmaceutical's mechanism of action and is easily implemented by quality control (i.e., must be a validated assay) can present major challenges for biopharmaceutical product development. Dr. Lankford will discuss regulatory considerations associated with the development and validation of an appropriate potency assay and potency assay expectations for different phases of clinical development and licensure.
Carla S.R. Lankford, M.D., Ph.D., Product Quality Reviewer, Division of Monoclonal Antibodies, Office of Biotechnology Products, CDER, US FDA
Biologics license applications must demonstrate that the product is safe, pure and potent according to the Public Health Service (PHS) Act 42, USC 262, and the Code of Federal Regulations (21 CFR), 601.2. It is emphasized in the PHS Act under Subpart 1-Biological Products, and 21 CFR 610.10-Potency that all biological drug products are required to be released using an assay that measures their potency. Development of a potency assay that is both reflective of the biopharmaceutical's mechanism of action and is easily implemented by quality control (i.e., must be a validated assay) can present major challenges for biopharmaceutical product development. Dr. Lankford will discuss regulatory considerations associated with the development and validation of an appropriate potency assay and potency assay expectations for different phases of clinical development and licensure.
Carla S.R. Lankford, M.D., Ph.D., Product Quality Reviewer, Division of Monoclonal Antibodies, Office of Biotechnology Products, CDER, US FDA
New, Unpublished Data
9:15
Case
Study
Potency Assay(s) for a Dual Action Fab Antibody in QCStudy
This case study will describe the strategy in the development and selection of a potency method(s) for a dual-action Fab (DAF), a monoclonal antibody able to recognize two different antigens. Validation studies will be presented.
Guoying Jiang, Ph.D., Associate Scientist, Analytical Development and Quality Control, Genentech, Inc.
9:45
New and Revised USP Bioassay Chapters
The US Pharmacopeia is replacing its current general chapter Design and Analysis of Biological Assays <111> with a suite of five chapters - Overview of USP Bioassay Chapters <1030>, Design and Development of Biological Assays <1032>, Validation of Biological Assays <1033>, Analysis of Biological Assays <1034>, and a substantially revised <111>. The purpose of this talk is to summarize the changes, their status, and impact of the changes on monographs and other chapters.
Walter W. Hauck, Ph.D., Senior Scientific Fellow, United States Pharmacopeia
The US Pharmacopeia is replacing its current general chapter Design and Analysis of Biological Assays <111> with a suite of five chapters - Overview of USP Bioassay Chapters <1030>, Design and Development of Biological Assays <1032>, Validation of Biological Assays <1033>, Analysis of Biological Assays <1034>, and a substantially revised <111>. The purpose of this talk is to summarize the changes, their status, and impact of the changes on monographs and other chapters.
Walter W. Hauck, Ph.D., Senior Scientific Fellow, United States Pharmacopeia
10:15
Networking Refreshment Break and Exhibit/Poster Viewing
New, Unpublished Data
10:45
Case
Study
Challenges in the Development of a Bioassay for a Therapeutic Antibody Targeting Cancer Stem CellsStudy
In this study, we describe the development of a reporter gene bioassay used to measure the potency of our cancer stem cell targeting antibody. To improve the variability, we developed creative methods to limit the number of steps and cell lines used in the bioassay. In our study, we describe the challenges encountered in developing a bioassay for this antibody that can be transferred and used in a QC environment.
Esohe Idusogie, Ph.D., Senior Scientist and Group Leader, Oncomed Pharmaceuticals
New, Unpublished Data
11:15
Case
Study
Correlation between Binding and Cell-based Functional Assays for the Potency of Monoclonal Antibody ProductsStudy
Biogen Idec has developed monoclonal antibody products against multiple target proteins. In order to examine the potency of these antibodies, both binding and cell-based functional assays have been developed for each product. It is important to examine correlation between these potency assays to determine the best assays to be used for lot release, stability studies and product characterization. This presentation will discuss strategies for carrying out correlation studies and present data from different products.
Wei Zhang, Ph.D., Senior Scientist, Analytical Development, Biogen Idec
11:45
Q&A Panel Discussion with Morning Speakers
12:15
Networking Luncheon and Exhibit/Poster Viewing
1:25
Chairperson's Remarks
Michael G. Mulkerrin, Ph.D., Vice President, Process Development and Manufacturing, OncoMed Pharmaceuticals
Michael G. Mulkerrin, Ph.D., Vice President, Process Development and Manufacturing, OncoMed Pharmaceuticals
Post-Translational Modifications
New, Unpublished Data
1:30
Case
Study
Deamidation of Asparagine Residues in Recombinant IgG mAb's Results in a Cluster of Multiple Reaction ProductsStudy
A recombinant IgG mAb was subjected to tryptic LC-MS peptide mapping studies and of 28 potential sites of deamidation, 6 were identified as having multiple deamidation products. Two of these sites have the most susceptible NG motif, but the others have less prone motifs and still demonstrate multiple reaction products. In-depth study of one of these product clusters was performed using synthetic standards to assign uneqivocal identity to each species as mass and fragmentation cannot discriminate stereoisomers. The existence of these deamidation product clusters has implications in the amount of deamidation reported for biologics.
Adam Lucka, Scientist II, Department of Protein Characterization, Alexion Pharmaceuticals, Inc.
New, Unpublished Data
2:00
Case
Study
Identification and Characterization of rhuMab A Variants with Buried Unpaired CysteinesStudy
Antibody disulfide linkage and free thiols are important quality attributes. Here we will present the characterization of variants with buried unpaired cysteines for rhuMab A. We observed several chromatographic heterogeneities for rhuMab A. Under denaturing conditions, the Ellman's assay revealed the presence free thiols. LC-MS showed the unpaired cysteines are cys 22 and cys 96 in the heavy chain variable domain. The enriched variants' activities will be discussed.
Taylor Zhang, Ph.D., Scientist, Protein Analytical Chemistry-1, Genentech, Inc.
New, Unpublished Data
2:30
On-Column Conversion of IgG1 Trisulfide Linkages to Native Disulfides in Tandem with Protein A Affinity Chromatography
Molecular heterogeneity was detected in a recombinant IgG1 mAb and attributed to the presence of a protein trisulfide moiety. The trisulfide was eliminated during routine purification of the IgG1 mAb via a REDOX wash step incorporated into Protein A affinity column chromatography. Similar, high efficiency trisulfide conversion was achieved for a second IgG1 mAb and a novel IgG1-based molecule using the column wash strategy. Therefore, trisulfide/disulfide heterogeneity may be eliminated from IgG1 molecules via a convenient, controllable and inexpensive procedure compatible with routine Protein A affinity capture.
David Evans, Ph.D., Scientist, Process Biochemistry, Biogen Idec
Molecular heterogeneity was detected in a recombinant IgG1 mAb and attributed to the presence of a protein trisulfide moiety. The trisulfide was eliminated during routine purification of the IgG1 mAb via a REDOX wash step incorporated into Protein A affinity column chromatography. Similar, high efficiency trisulfide conversion was achieved for a second IgG1 mAb and a novel IgG1-based molecule using the column wash strategy. Therefore, trisulfide/disulfide heterogeneity may be eliminated from IgG1 molecules via a convenient, controllable and inexpensive procedure compatible with routine Protein A affinity capture.
David Evans, Ph.D., Scientist, Process Biochemistry, Biogen Idec
3:00
Networking Refreshment Break and Exhibit/Poster Viewing
New, Unpublished Data
3:30
Case
Study
A Tale of Three Formulations: Challenges and Strategies in the Development of a Late Stage Monoclonal AntibodyStudy
The development of a fully human, affinity-matured IgG1 monoclonal antibody for late stage clinical trials and final commercial market presented numerous formulation challenges. An increase in aggregates was linked to excipient crystallization in the frozen state and was alleviated by replacing the cryoprotectant. To diminish potential oxidation risks in the final marketed product, two distinct formulations, a solid and a liquid, were developed. Strategies to decrease reconstitution time in the lyophilized drug product, formulate a challenging IgG1 as a liquid formulation, and introduce new formulation excipients during manufacturing will be discussed.
Yvonne Lentz, Ph.D., Scientist, Late Stage Pharmaceutical and Processing Development, Genentech, Inc.
4:00
Q&A Panel Discussion with Afternoon Speakers
Special Joint Session
Regulatory Perspectives and "Pet Peeves"
4:15
Pet Peeves and Roadblocks in CMC: Improving your Quality Submissions through the Product Lifecycle
Avoiding common mistakes in submissions to the FDA leads to a smoother developmental pathway for your therapeutic protein. Roadblocks to development during the product lifecycle and case studies will be presented. A collection of Office of Biotechnology reviewer pet peeves will be shared.
Chana Fuchs, Ph.D., Team Leader, Division of Monoclonal Antibodies, CDER, US FDA
Marjorie Shapiro, Ph.D., Chief, Laboratory of Molecular & Developmental Immunology, CDER, US FDA
Avoiding common mistakes in submissions to the FDA leads to a smoother developmental pathway for your therapeutic protein. Roadblocks to development during the product lifecycle and case studies will be presented. A collection of Office of Biotechnology reviewer pet peeves will be shared.
Chana Fuchs, Ph.D., Team Leader, Division of Monoclonal Antibodies, CDER, US FDA
Marjorie Shapiro, Ph.D., Chief, Laboratory of Molecular & Developmental Immunology, CDER, US FDA
5:15
Q&A Panel Discussion with FDA Speakers
5:30
Close of Day Two
8:15
Coffee
8:15
Chairperson's Remarks
Brian B. Lentrichia, Ph.D., Associate Director, Pharmaceutical & Analytical Development, Shire Human Genetic Therapies
Brian B. Lentrichia, Ph.D., Associate Director, Pharmaceutical & Analytical Development, Shire Human Genetic Therapies
Vaccine Characterization
8:30
Analytical Characterization for Gene Therapy Products
Michael Havert, Ph.D., Biologist, Division of Cell and Gene Therapies, CBER US FDA
Michael Havert, Ph.D., Biologist, Division of Cell and Gene Therapies, CBER US FDA
New, Unpublished Data
9:00
Case
Study
Biophysical and Biochemical Characterization of Insect Cell Produced Virus-Like Particle (VLP)VaccinesStudy
A unique feature of our VLPs (unlike existing VLP vaccines) is that they are membrane containing structures in which are inserted three influenza proteins HA, NA and Matrix protein. I will describe how Novavax produces and purifies VLPs and has been characterizing these particles in terms of membrane fatty acids and lipids, host proteins, and carbohydrate and data demonstrating a lack of aggregation toward our eventual goal of having a well characterized biological. I will also describe some ways that these particles are different from the baculovirus vectors that are used to infect the cell and initiate their production.
Steven Pincus, Ph.D., Executive Director, Analytical Operations, Novavax, Inc.
9:30
Conjugation Impurities and Carrier Protein Characterization of a Therapeutic Vaccine
Therapeutic vaccines are complex bioconjugates having many components, including a carrier protein, linkers, and haptens. A common approach for hapten conjugation is the use of heterobifunctional linkers, which results in multiple sites of attachment onto the carrier protein. The carrier protein also contains many inherent characterization challenges, including uncertainty in the theoretical amino acid sequence, glycosylation complexity, and unique thioether modifications. This presentation describes a combination of analytical approaches to characterize the conjugation impurities and carrier protein of a therapeutic vaccine.
Justin B. Sperry, Ph.D., Senior Scientist, BioTherapeutics Research and Development, Pfizer, Inc.
Therapeutic vaccines are complex bioconjugates having many components, including a carrier protein, linkers, and haptens. A common approach for hapten conjugation is the use of heterobifunctional linkers, which results in multiple sites of attachment onto the carrier protein. The carrier protein also contains many inherent characterization challenges, including uncertainty in the theoretical amino acid sequence, glycosylation complexity, and unique thioether modifications. This presentation describes a combination of analytical approaches to characterize the conjugation impurities and carrier protein of a therapeutic vaccine.
Justin B. Sperry, Ph.D., Senior Scientist, BioTherapeutics Research and Development, Pfizer, Inc.
10:00
Networking Refreshment Break and Exhibit/Poster Viewing
Testing and Characterization of Host Cell Impurities
New, Unpublished Data
10:30
Case
Study
Detection and Quantitation of Host Cell Proteins in Biopharmaceutical ProductsStudy
Host cell proteins (HCP) are the major protein contaminants in biopharmaceutical products and they can affect therapeutic efficacy and safety. HCPs are typically measured using enzyme-linked immunosorbent assays (ELISAs) with polyclonal antibodies that are capable of detecting a broad range of HCPs. At ImClone, we generate polyclonal antibodies targeting several CHO host cell proteins and monitor their presence in the monoclonal antibody products using Chemiluminescence ELISA, one and two-dimensional SDS-PAGE followed by protein identification using Western blotting.
Shanmuuga Sundaram, Ph.D., Senior Scientist, BioAnalytical Science, ImClone Systems
New, Unpublished Data
11:00
Is There a Current Technology that Stands Out When It Comes to Host Cell Protein Analysis?
Assay performance, sample analysis and assay complexity was monitored on the following technologies: plate based colorimetric ELISA, plate based electro-chemiluminescence (ECL) ELISA, a bead based luminescence assay, and a homogeneous time resolved fluorescence (FRET) assay, to determine how each of these technologies improved HCP analysis and time to result.
Martha Jackson, Senior Research Scientist I, Pfizer, Inc.
Assay performance, sample analysis and assay complexity was monitored on the following technologies: plate based colorimetric ELISA, plate based electro-chemiluminescence (ECL) ELISA, a bead based luminescence assay, and a homogeneous time resolved fluorescence (FRET) assay, to determine how each of these technologies improved HCP analysis and time to result.
Martha Jackson, Senior Research Scientist I, Pfizer, Inc.
11:30
Analytical Challenges with Determination of HCP Clearance during Purification of the Product Expressed in a Human Host Cell System
Over expression of recombinant lysosomal enzymes from a human host cell offers the benefit of a well-tolerated human glycosylation pattern. However, monitoring clearance of human proteins from an endogenous human target product using a generic HCP ELISA presents a dual challenge; meeting system suitability when quantifying residual impurities in the presence of a high concentration of target protein and coping with possible anti-product reactivity. Purification changes the relative level of HCP through the process and can reveal the limitations of an assay best suited to a complex mixture of species. The amelioration of the assay requires the identification of highly immunoreactive species in highly pure samples by HCP ELISA and a sensitive HCP Western blot. We present a strategy of re-formatting a generic human HCP ELISA to overcome the limitations of the assay for application to a human host cell system.
Brian B. Lentrichia, Ph.D., Associate Director, Pharmaceutical & Analytical Development, Shire Human Genetic Therapies
Over expression of recombinant lysosomal enzymes from a human host cell offers the benefit of a well-tolerated human glycosylation pattern. However, monitoring clearance of human proteins from an endogenous human target product using a generic HCP ELISA presents a dual challenge; meeting system suitability when quantifying residual impurities in the presence of a high concentration of target protein and coping with possible anti-product reactivity. Purification changes the relative level of HCP through the process and can reveal the limitations of an assay best suited to a complex mixture of species. The amelioration of the assay requires the identification of highly immunoreactive species in highly pure samples by HCP ELISA and a sensitive HCP Western blot. We present a strategy of re-formatting a generic human HCP ELISA to overcome the limitations of the assay for application to a human host cell system.
Brian B. Lentrichia, Ph.D., Associate Director, Pharmaceutical & Analytical Development, Shire Human Genetic Therapies
12:00
Q&A Panel Discussion with Morning Speakers
12:30
Lunch on your own
1:30
Chairperson's Remarks
Flaviu Gruia, Ph.D., Scientist I, Analytical Biochemistry, MedImmune, Inc.
Flaviu Gruia, Ph.D., Scientist I, Analytical Biochemistry, MedImmune, Inc.
Monitoring Protein Aggregation and Particulate Formation
New, Unpublished Data
1:45
Case
Study
Nanosight - A Novel Approach for Sub-micron Particle Detection and AnalysisStudy
Nanosight is a promising emerging technique that combines elements of light scattering, microscopy and particle tracking software in order to accurately describe sub-micron particle distributions in liquid solutions. Advantages, limitations and recent applications of Nanosight will be presented, with extra focus on submicron particle distributions as an early predictor for the later appearance of visible and sub-visible particles. Newer developments in this area and how they could affect future capabilities of this technique will also be discussed.
Flaviu Gruia, Ph.D., Scientist I, Analytical Biochemistry, MedImmune, Inc.
2:15
Challenges in Measuring Comparability of Aggregation and Conformation for Biosimilars and Innovative Biological Products
Biosimilar developers do not have access to the innovator's bulk drug substance (BDS), and this often creates problems for key biophysical tools such as AUC, light scattering, and CD due to interference from excipients. We show that trying to re-purify BDS from the innovator's final product can lead to invalid comparisons. Some specifics about excipient interference with AUC, light scattering, CD, and/or SEC will be discussed, with examples.
John Philo, Ph.D., Vice President and Director of Biophysical Chemistry, Alliance Protein Laboratories
Biosimilar developers do not have access to the innovator's bulk drug substance (BDS), and this often creates problems for key biophysical tools such as AUC, light scattering, and CD due to interference from excipients. We show that trying to re-purify BDS from the innovator's final product can lead to invalid comparisons. Some specifics about excipient interference with AUC, light scattering, CD, and/or SEC will be discussed, with examples.
John Philo, Ph.D., Vice President and Director of Biophysical Chemistry, Alliance Protein Laboratories
2:45
Networking Refreshment Break and Exhibit/Poster Viewing
Comparability Case Studies
3:15
Case
Study
Comparability of Biotechnology Products: Regulatory Considerations and Case StudiesStudy
Abstract not available at time of print.
Rashmi Rawat, Ph.D., Product Quality Reviewer, Division of Monoclonal Antibodies, CDER, US FDA
3:45
Case
Study
Comparison of Multiple Lots of a Chimeric Monoclonal Antibody Manufactured over a Ten Year PeriodStudy
Multiple lots of a chimeric monoclonal antibody for the treatment of pediatric neuroblastoma have been produced over ten years for an NCI-Sponsored non-pivotal Phase III trial. The product is produced in an SP2/0 cell line using hollow-fiber technology. Physicochemical and biological assays were utilized to demonstrate comparability between lots. Despite changes in the distribution of the glycan structure there was no detectable change in in vitro functionality.
Steven Giardina, Ph.D., Director, Process Analytics, Biopharmaceutical Development Program, SAIC-Frederick, Inc.
4:15
Q&A Panel Discussion with Afternoon Speakers
4:45
Close of Conference
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