To whom correspondence should be addressed.
Chapter 10. IFN-γ ELISPOT Assay Validation
- Uma Prabhakar Ph.D. and
- Marian Kelley Director of Compliance
Published Online: 8 APR 2008
Copyright © 2008 John Wiley & Sons, Ltd
Validation of Cell-Based Assays in the GLP Setting: A Practical Guide
How to Cite
Reddy, M., Wong, J., Pendley, C. and Prabhakar, U. (2008) IFN-γ ELISPOT Assay 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.ch10
Department of Clinical Pharmacology & Experimental Medicine, Centocor Research and Development, Inc., 145 King of Prussia Road, Radnor, PA 19087, USA
- Published Online: 8 APR 2008
- Published Print: 21 MAR 2008
Print ISBN: 9780470028766
Online ISBN: 9780470987810
- peripheral blood mononuclear cells;
The utility of the IFN-γ ELISPOT assay for detecting the frequency of antigen-specific T lymphocytes in autoimmune disease models and for immune responses in patients with infectious, autoimmune neoplastic diseases undergoing immunotherapeutic treatments has been reported in several studies. Development, optimization, validation of an IFN-γ ELISPOT assay for use in the analysis of clinical study samples will be described in this chapter. Several assay conditions such as cell number, length of culture, positive and negative controls, and antigen concentration will be addressed. Procedures validating the stability of responses in shipped samples, stability assessment of long term cryopreserved samples, assay precision including intra-assay (variability in replicates), inter-assay (plate-to-plate variability and within day variability), inter-day and inter-operator variability's will also be described. In addition, data analysis and interpretation including lower and upper limits of quantification will be discussed.