Validation of Cell-Based Assays in the GLP Setting

The use of cell-based assays within pharmaceutical and biotechnology companies is driven in large part by the need to evaluate the plethora of drug targets derived from genomics and proteomics. In addition, the potential of biomarkers to facilitate the development of effective and safe drugs is being recognized as an integral part of all phases of drug development, and cell-based technologies are a critical part of biomarker discovery and development. Despite this critical role, cell-based assays have not been standardized and made compliant with Good Laboratory Practice guidelines.   In this book, the editors have collected assays for which validation procedures have been developed, making this a vital purchase for anyone using such assays in drug development. This book:  Describes the development, optimization and validation of cell-based assays, including procedural documentation required for Good Laboratory Practice Presents validations of cell-based assays for select targets, with step-by-step instructions, allowing the reader to reproduce the assay conditions and results Provides details of techniques used in the evaluation of immunodeficiency, autoimmune and oncological disorders, including assessment of cancer vaccines Offers a compendium of validation parameters that need to be considered when using these methods to develop a new drug Includes detailed protocols for the evaluation of cytokines and of neutralizing antibodies directed against protein therapeutics Validation of Cell-based Assays in the GLP Setting provides the professional with an invaluable reference source, featuring key guidelines. The book will prove extremely useful to all scientists working in the areas of drug development.

List of contributors ix Preface xiii Introduction xvii Uma Prabhakar 1 Considerations while setting up cell-based assays 1 Marian Kelley 2 Development, optimization and validation of cell-based assays – 1 11 Marielena Mata and Thomas Lohr 3 Development, optimization and validation of cell-based assays – 2 25 Manjula Reddy and Uma Prabhakar 4 Whole blood ex vivo stimulation assay development, optimization and validation 37 Manjula Reddy and Uma Prabhakar 5 Immunohistochemistry assays in Good Laboratory Practice studies 49 Frank Lynch, Steve Bernstein and Hector Battifora 6 Flow cytometric cell-based assays: an overview of general applications 73 Cuc Davis, Manjula Reddy, Thomas Williams and Uma Prabhakar 7 T-cell surface markers in human peripheral whole blood using flow cytometry 85 Manjula Reddy, Cuc Davis, Hugh Davis, Charles Pendley and Uma Prabhakar 8 Intracellular cytokine detection by flow cytometry 107 Julie G. Wilkinson, Carlos A. Aparicio and Wade E. Bolton 9 Validating reference samples for comparison in a regulated ELISPOT assay 127 Magdalena Tary-Lehmann, Christina D. Hamm and Paul V. Lehmann 10 IFN-γ ELISPOT assay validation 147 Manjula Reddy, Jackson Wong, Charles Pendley and Uma Prabhakar 11 IL-5 ELISPOT assay validation 173 Manjula Reddy, Jackson Wong, Hugh Davis, Charles Pendley and Uma Prabhakar 12 Validation of the Cylex technology to measure T and B cell activation capacity in clinical trials 193 Marielena Mata, Thomas Lohr and Jaymala Patel 13 Development of validated neutralization bioassays 209 Manoj Rajadhyaksha, Manjula Reddy, Jaime Bald, Amy Fraunfelter, Persymphonie Miller, Marian Kelley and Uma Prabhakar 14 Endpoint assays in HIV-1 vaccine trials: functioning in a Good Laboratory Practices environment 239 Patricia D’Souza, Josephine H. Cox, Guido Ferrari, Nina Thapa Kunwar, Victoria Polonis and Marcella Sarzotti-Kelsoe 15 The future direction of cell-based assays 277 Uma Prabhakar and Marian Kelley Index 283