Chapter 4. Whole Blood Ex vivo Stimulation Assay Development, Optimization and Validation

  1. Uma Prabhakar Ph.D. and
  2. Marian Kelley Director of Compliance
  1. Manjula Reddy and
  2. Uma Prabhakar Ph.D.

Published Online: 8 APR 2008

DOI: 10.1002/9780470987810.ch4

Validation of Cell-Based Assays in the GLP Setting: A Practical Guide

Validation of Cell-Based Assays in the GLP Setting: A Practical Guide

How to Cite

Reddy, M. and Prabhakar, U. (2008) Whole Blood Ex vivo Stimulation Assay Development, Optimization and Validation, in Validation of Cell-Based Assays in the GLP Setting: A Practical Guide (eds U. Prabhakar and M. Kelley), John Wiley & Sons, Ltd, Chichester, UK. doi: 10.1002/9780470987810.ch4

Editor Information

  1. Department of Clinical Pharmacology & Experimental Medicine, Centocor Research and Development, Inc., 145 King of Prussia Road, Radnor, PA 19087, USA

Author Information

  1. Department of Clinical Pharmacology & Experimental Medicine, Centocor Research and Development, Inc., 145 King of Prussia Road, Radnor, PA 19087, USA

  1. To whom correspondence should be addressed.

Publication History

  1. Published Online: 8 APR 2008
  2. Published Print: 21 MAR 2008

ISBN Information

Print ISBN: 9780470028766

Online ISBN: 9780470987810

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Keywords:

  • whole blood;
  • ex vivo

Summary

Ex vivo cytokine secretion assays, where a clinical peripheral whole blood sample is exposed to a therapeutic agent and a resulting response measured, have proven to be a powerful tool for assessing cellular responses. Various sources contribute to the variability within an ex vivo stimulation assay. Therefore, optimization of conditions for blood collection, storage, and processing of an ex vivo cytokine stimulation assay are important. The optimized and validated conditions should result in an assay that shows operational feasibility, minimal imprecision, elimination of potential confounding factors and should be robust enough for application to clinical drug development. In this chapter, we will present guidelines for the development of validation plans for ex vivo immunoassays used to evaluate cellular immune responses using whole blood (WB) samples obtained from non-clinical and clinical studies.

In the cases where a resulting cytokine is typically measured, the use a commercial kit is often required. Kit qualification/validation has been discussed thoroughly in previous papers and will not be addressed in this chapter. The focus of this discussion will be on the validation of the cellular aspect of the assay method, leading up to the end response.