Biological Assay Development, Validation & Maintenance
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Workshop #1:
Advanced Tools for Assessing and Controlling Assay Variability and Spurious OOS Results
Workshop Leader:
Part 1: Managing Variability in the Bioassay
This workshop will combine concepts with hands-on exercises to reinforce the implementation of the design and analysis tools. Timothy Schofield has recently joined Arlenda, a statistical consulting and software company providing support to the pharmaceutical, devices, and analytical testing industries. Prior to joining Arlenda Tim was Director in the U.S. Regulatory Affairs department of GlaxoSmithKline where he provided regulatory support to vaccines, as well as head of the Nonclinical Statistics department in Merck Research Labs, supporting development and manufacture of Merck pharmaceuticals, biologics, and vaccines. In addition to his service to GSK and Merck, Tim is co-chair of the USP Statistics Expert Committee and a member of the USP Bioassay Validation ad hoc panel where he led efforts to write Chapter <1033> Bioassay Validation.
Part 2: Approaches to Determining and Minimizing OOS from Bioassays
David Lansky's educational background includes a year at the University of Michigan in Electrical and Computer Engineering, a B.S. in Botany from San Francisco State University, followed by three graduate degrees from Cornell: an M.S. in Entomology then an M.S. and Ph.D. in Biometry. David has worked on bioassay development, analysis and validation for over 20 years. Most of his experience is in industry, including 10 years at Searle/Monsanto/Pharmacia and nine years as the owner and lead consultant at Precision Bioassay, Inc., a consulting firm focused on statistical methods and software for bioassay. |
Workshop #2:
Bioassay Development Basics 101
Workshop Leaders: Biological assays can be precise and easy to perform. This bioassay development workshop will begin with an overview of the basic tools required for success: analyst training, critical reagent maintenance, laboratory/equipment set-up and regulatory expectations for Phase I/I/III clinical trials. Practical approaches to designing assay formats, system suitability, and preparation for bioassay transfer will be discussed and case studies will be presented.
Michael Merges is Director of Catalent's Biopharmaceutical Support Services (BPS). BPS's focus is the transfer, development, validation, and performance of bioassays. Michael has experience with many techniques including cell-based (primary cells and cell lines) bioassays, immunological/neutralization assays, ELISA, and Flow Cytometry. He joined Catalent from Lonza where he was Associate Director of Bioservices. Prior to that, he was at the University of Maryland's Institute of Human Virology where he served as the Institute's Research Supervisor. He has also conducted viral immunology research at the National Cancer Institute and Johns Hopkins University. He obtained his Bachelor's Degree in Microbiology from The Pennsylvania State University and his Master's Degree in Microbiology/Virology from Hood College. Michael Sadick received his B.A. in biology from Johns Hopkins, in Baltimore, and his MS and Ph.D. in immunology from University of Washington in Seattle. He worked as research faculty at UCSF Medical Center for five years. Following his work at UCSF Medical Center, Michael then worked for 10 years (1991 - 2001) at Genentech in South San Francisco as a Senior Scientist in the Bioassay Group supporting Pharmaceutical Science efforts (Phases I - III). Michael was recruited to Eli Lilly and Co. as a Research Advisor in 2001 to help lead biotechnology efforts, leading the Bioassay Groups, Molecular Biology and Virology in support of Phase I-III projects, as well as providing guidance for commercial bioassay testing, all on a global level, working with FDA and EMEA regulatory agencies. In 2007, Michael joined Aptuit, a CRO in Kansas City, MO, as a Senior Manager in the Large Molecule Analysis and Characterization (LMAC) division. |
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Schedule for Both Workshops
8:00
Registration and Coffee
9:00
Workshops Begin
10:30-11:00
Networking Refreshment Break with Poster and Exhibit Viewing
12:30-1:30
Luncheon in Poster and Exhibit Hall
1:30
Workshops Resume
3:30-4:00
Networking Refreshment Break with Poster and Exhibit Viewing
5:00
Close of Workshop
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ワークショップ: 月曜日 | メインカンファレンス: 火曜日 | 水曜日
Laureen E. Little, Ph.D., Principal Consultant, Quality Services; Editor and Publisher, BioQuality
Featured Presentation
The USP chapters on biological assay development, validation, and analysis have undergone final review prior to publication in USP NF. Comments received from industry and FDA have been addressed in the final chapters. This talk will give an overview of the chapters, highlighting both the practical and statistical recommendations. The chapters will be contrasted to other regulatory guidances on analytical methods.
Timothy Schofield, M.A., Managing Director, Arlenda USA
Dealing with Animal Potency Assays
The animal potency assay was designed and refined to balance the dynamic range, product specification, and number of animals used. In addition, the assay variability was identified and controlled.
Hongran Stone, Ph.D., Senior Director, Revance Therapeutics, Inc.
An assessment of vaccine potency is necessary to predict clinical effectiveness from lot to lot and insure safety and efficacy. Assays designed to access vaccine potency are by necessity indirect and often rely on in vivo animal models or in vitro bioassays. The challenge is establishing a correlate between the in vitro and in vivo models that effectively addresses the mechanism of action and is acceptable to the regulatory agencies. This is compounded by the complexity of the vaccine that may contain multiple antigens in addition to an excipient or adjuvant. Animal models that respond to the vaccine are ideal; however in vitro assessment is necessary to routinely evaluate manufacturing consistency, stability, and strength. An approach to an in vitro potency assay is discussed in relation to the challenges of establishing an in vivo correlate, stability indicating properties, and vaccine complexity.
Leonard Blackwell, Ph.D., Principal Scientist, BioTherapeutics Pharmaceutical Sciences, Pfizer
Abstract unavailable at press date.
Jamie Bird, M.Sc., Supervisor, Emergent BioSolutions
Hear a case study of validation of a multiplex serology assay (DTP-6 IgG) designed to simultaneously measure IgG antibody levels to six DTaP vaccine antigens serum samples. The DTaP antigens included in the assay are PTx, FHA, PRN, and FIM 2/3 of Bordetella pertussis, DTd of Corynebacterium diphtheriae, and TTd of Clostridium tetani. Validation parameters that were assessed included sensitivity, limits of quantitation, ruggedness, precision, dilutability, specificity, selectivity and accuracy.
Katie Matys, M.S., Research Scientist, PPD Vaccines and Biologics Lab
The Evolving Art of Measuring Antibodies in Serum
Bioassay or blocking assays are commonly used to determine the ability of Anti-Drug Antibody (ADA) to neutralize biological effect of the drug. To determine whether ADA blocked binding of drug to the receptor or ligand expressed on cell surface, Flow ctyometry and ECL based methods were employed during clinical development of two biotherapeutics. This presentation will cover the challenges during implementation of such bioassays in routine clinical testing.
Jaya Goyal, Ph.D., Principal Investigator, Biogen Idec
Immunogenicity concerns for biologics necessitate the need for functional assays to detect neutralizing antibodies in patient matrix that can impact safety and efficacy. While cell-based assays are the preferred format for neutralizing antibody (NAb) assays, they present unique technical challenges due to the effect of matrix components. A case study that highlights some of the challenges faced in the development of a NAb assay will be presented.
Renuka C. Pillutla, Ph.D., Associate Director, Bristol-Myers Squibb
Potency Assays for Multi-Functional Antibody Products
Xu-Rong Jiang, M.D., Ph.D., Associate Director, MedImmune
The presentation will provide an overview of the white paper on Fc effector function of therapeutic antibodies published in February 2011 (Nature Reviews Drug Discovery 10, 101-111). A summary of the current knowledge of antibody Fc functionality, a strategy for assessing the effector functions of different classes of therapeutic antibodies (including Fc fusion proteins) and a proposed path for routine testing and controls for manufacturers of antibody products will be covered.
Svetlana Bergelson, Ph.D., Associate Director, Biogen-Idec
Case studies for the set up and optimization of functional assays such as Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) as well as a Compliment-Dependent Cytotoxicity (CDC) for therapeutic antibodies will be presented. In addition to these MOA assays, the set up and validation of a flow cytometric binding assay according to ICH Q2(R1) guidelines will be presented using Rituximab as an example.
Ulrike Herbrand, Ph.D., Scientific Officer, Bioassays, Charles River Biopharmaceutical Services GmbH, Germany
Abstract unavailable at press date.
Xu-Rong Jiang, M.D., Ph.D., Associate Director, MedImmune
Abstract unavailable at press date.
Sonia Connaughton, Ph.D., Senior Scientist, ImmunoGen
Abstract unavailable at press date.
Jun Kim, Associate Scientist, MedImmune
ワークショップ: 月曜日 | メインカンファレンス: 火曜日 | 水曜日
Sally Seaver, Ph.D., President, Seaver Associates LLC
Featured Presentations
A bioassay (or an in vitro potency assay) is a critical component of the control systems that monitor consistency and stability of a manufactured biological product. Potency assays used at early stages of product development may not be the same as those used during late stage product development or for licensure - often there is a need to modify or change the assay to meet additional FDA requirements. This presentation is aimed at providing an overview of some of the Agency observations, experiences and expectations regarding bioassay development over the last decade.
Gerald Feldman, Ph.D., Research Biologist, CDER, US FDA
Bioassay designs are constrained by practical considerations, statistical principles, and variation in biological materials. Good design, lab technique, and analyses combine to yield high performance bioassays. Recent simulations illustrate the impact of design and analysis on bioassay performance over useful ranges of potency and variation (both within and between assays).
David Lansky, Ph.D., President, Precision BioAssay
How Consistent is your Assay?
Enzyme-linked immunosorbent assays (ELISAs) are commonly used in the Quality Control of biopharmaceuticals. Their application includes the control of the depletion of process related impurities as well as the identification, quantitation or potency determination of biological test substances. This talk provides assistance for defining key parameters in assay development, evaluation of assay performance and method validation by considering specifically the intended use of the assay to fulfill scientific and regulatory requirements.
Thomas Waerner, Ph.D., Head Cell & Molecular Biology, Boehringer Ingelheim RCV GmbH & Co KG, Germany
Abstract unavailable at press date.
Kevin Brooks, Ph.D., Biostatistician, Emergent BioSolutions
During the life of a product the potency assay goes through progressive phases from development to the validation of its use for a release assay. These phases usually occur in different laboratories, with a transfer of the assay from the development laboratory to the manufacturing laboratory. To ensure a successful transfer, it is important that both laboratories obtain comparable assay results. We used a blinded panel of specimens and common critical reagents to perform assay testing in three laboratories involved in assay transfer. Concordance, Altman-Bland Agreement and Precision analyses were performed on quantifiable assay results.
Janet L. Lathey, Ph.D., Director, Emergent BioSolutions
Assay Components: The Devil is in the Details
Clinical laboratories utilize various reference standards during testing for potency assays. Two types of references are used: International reference standards or in-house standards. The international reference standards are bought from WHO, CBER, NIBSC, etc. Very often clinical laboratories prepare new reference lots to be used for testing because the lot is depleted or has reached the expiration date. In order to be used, the new reference needs to be re-characterized, i.e., the potency of the new lot shall be within specifications when compared to the old lot, the reference standard, to determine whether it is appropriate to switch the bioassay standard lot to the new lot. This paper describes reference recharacterization study design for potency assay using four-parameter logistic model.
Eloi Kpamegan, Ph.D., MSF, Director, Novavax, Inc.
Our BioAnalytical Assays department is currently evaluating various assay maintenance methods, activities and practices in an effort to better harmonize initial assay validation with long-term assay maintenance, and to better characterize assay performance over time. It is therefore important that analytical methods function both accurately and consistently. The following is a test case which demonstrates an alternative approach to experimental set-up, data analysis, and result interpretation when performing the following assay maintenance activities: 1) Reagent incorporation. 2) Long term reagent stability determination. 3) Monitoring long term assay performance. 4) Anticipating possible future performance issues based upon past data.
Bartek Bossak, Research Associate, Genentech, Inc.
The quality of reagents dictates the performance of cell-based assays and control of these reagents is important to ensure the reproducibility and consistency of the assay. The qualification of reagents used in serological neutralization assays, including viruses, cells, fetal bovine serum, cell culture medium, primary antibodies, secondary antibodies, and assay quality controls are described. The methods discussed here have been successfully applied to a variety of serological neutralization assays using viruses such as paramyxoviruses, orthomyxoviruses and flaviviruses.
Tatyana Timiryasova, Ph.D., Scientist, Sanofi Pasteur, Inc.
Technology Workshop
We have developed an alternative bioassay to quantify potency of Fc effector function in ADCC of monoclonal antibodies bound to target cells. The bioluminescent reporter assay has good precision and accuracy, and is stability -indicating. It differentiates well between biological activities of preparations of antibody drug with small differences in glycosylation that affect Fc effector activity in ADCC.
Terry Surowy, Ph.D., Research Manager, Promega Corporation
Developing MultiProduct Host Cell Protein Assays
Host cell proteins (HCPs) are among the major biomanufacturing process-related impurities and are considered as CQA (critical quality attributes) by regulatory agencies. HCPs must be closely monitored for downstream purification steps and final drug product to ensure process consistency and product safety. Although ELISA has been the most common method for HCP quantification, one of the major drawbacks of HCP ELISA is assay accuracy since no single available ELISA assay kit has demonstrated to contain antibodies capable of detecting all the HCPs in CHO based biologics production process. In this presentation, orthogonal methods such as 2D-silver/western, 2D-DIGE and LCMSMS were used to achieve comparative study of identity of HCP standard used in Cygnus HPC ELISA (Cygnus 3G) with HCPs from three in-house null CHO cell lines.
Rong-Rong Zhu, Senior Scientist, EMD Millipore
At Biogen Idec, Host Cell Proteins (HCPs) are measured with in-house multi-product HCP assays using the ECL MSD platform. Presented here is our assay development strategy and a case study of an issue encountered during qualification of several products produced in Chinese Hamster Ovary (CHO) cells using the in-house assay.
Catherine Ramsey, Ph.D., Scientist, Biogen Idec