Prabhakar U., Kelley M. (Eds.) Validation of Cell-Based Assays in the GLP Setting: A Practical Guide

Chichester: John Wiley & Sons. – 2008. – 301 p. This book contains procedural documents we have developed to describe factors that should be taken into general consideration while setting up cellbased assays, and for the development, optimization and validation of cell assays. A number of actual validations are presented including ELISPOT, flow metric analysis, proliferation and neutralization of immune responses. We also have valuable contributions from several experts in the field who have provided their viewpoints for developing ELISPOT assays, intracellular cytokine assays, immunohistochemistry analysis, and endpoint assays for HIV-1 vaccine trials. Our incentive to publish this book is solely to provide the professional in the field examples of specific validations for complex cell-based assay platforms and their use in supporting clinical analysis of samples in a GLP setting. The editors do not claim that the methods and procedural documents presented here represent approved regulatory documents. Rather, they reflect best practices that should be followed to ensure consistent and reliable results along with good documentation practices. Our hope is that this book will serve as a living document and as new technology platforms become available, the procedures and practices are updated periodically. Continued efforts to reflect best laboratory practices will ensure the quality data at all times which in the long run will result in quality treatment for patients.Considerations while setting up cell-based assays
Development, optimization and validation of cell-based assays – 1
Development, optimization and validation of cell-based assays – 2
Whole blood ex vivo stimulation assay development, optimization and validation
Immunohistochemistry assays in Good Laboratory Practice studies
Flow cytometric cell-based assays: an overview of general applications
T-cell surface markers in human peripheral whole blood using flow cytometry
Intracellular cytokine detection by flow cytometry
Validating reference samples for comparison in a regulated ELISPOT assay
IFN-_ ELISPOT assay validation
IL-5 ELISPOT assay validation
Validation of the Cylex technology to measure T and B cell activation capacity in clinical trials
Development of validated neutralization bioassays
Endpoint assays in HIV-1 vaccine trials: functioning in a Good Laboratory Practices environment
The future direction of cell-based assays