Chapter 9. Validating Reference Samples for Comparison in a Regulated ELISPOT Assay

  1. Uma Prabhakar Ph.D. and
  2. Marian Kelley Director of Compliance
  1. Magdalena Tary-Lehmann MD, Ph.D.*,
  2. Christina D. Hamm and
  3. Paul V. Lehmann

Published Online: 8 APR 2008

DOI: 10.1002/9780470987810.ch9

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

Tary-Lehmann, M., Hamm, C. D. and Lehmann, P. V. (2008) Validating Reference Samples for Comparison in a Regulated ELISPOT Assay, 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.ch9

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. Cellular Technology Limited and Department of Pathology, Case Western Reserve University, Cleveland, OH 44106, USA

*Cellular Technology Ltd, 10515 Carnegie Ave., Cleveland OH 44106

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:

  • ELISPOT;
  • PBMC;
  • T cells

Summary

ELISPOT assays have recently emerged as a primary tool for monitoring antigen-specific low frequency measurements of cellular immunity, recording two cardinal features of cell-mediated immunity: the clonal sizes and the cytokine effector class. Obtaining information on these parameters is crucial for the comprehensive assessment, among other things, of any immunotherapy. This is particularly useful in the field of pre-clinical/clinical studies and vaccine trials, where high sample numbers from various sites have to be collected and stored before they can be processed in a timely fashion. With relatively little cell material and labor involved, ELISPOT assays would ideally lend themselves for high throughput analysis. While long employed in the academic research setting, the ability to obtain valid ELISPOT data on study samples is beginning to greatly enhance ex vivo immune monitoring capabilities for the pharmaceutical companies. To be used in a regulated environment, however, the assay needs to be validated. This chapter will focus on the standardization and validation of an ELISPOT assay using reference samples.