04/03/2009 11:36

CTL Writes Book Chapter on Validating Reference Samples for Use in Cell-Based Assays

In a new book on cell-based assays, CTL discusses key considerations in validating an ELISPOT assay.

Cellular Technology Limited has contributed a key chapter to the book, Validation of Cell-Based Assays in the GLP Setting: A Practical Guide. This text is one of the latest offerings by Research and Markets, a leading source for international market research and market data.

This book describes the development, optimization and validation of various cell-based assays, such as cytokine ELISPOT and ELISA assays, for which validation procedures have been developed. In addition, it discusses the validation of these assays for select targets using step-by-step instructions. Chapter 9 ("Validating reference samples for comparison in a regulated ELISPOT assay") was written by CTL personnel Magdalena Tary-Lehmann, Christina D. Hamm, and Paul V. Lehmann. In this chapter, they discuss the utility and performance of the ELISPOT method, as well as key considerations in validating such assays.

Validation of Cell-based Assays in the GLP Setting is an invaluable reference source for pharmaceutical companies and biotechnology firms that need to evaluate drug targets derived from genomics and proteomics. It is also a useful reference for companies that use cell-based technologies for biomarker discovery and development.

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.

 

For more information visit www.researchandmarkets.com/reports/c89231.


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CTL Serum-Free Media


Serum-free media allows users to standardize their cell culture conditions by avoiding the use of undefined and highly variable serum products derived from humans or animals, e.g. human AB serum or fetal calf serum (FCS).

The high variability in the biological properties of different serum batches makes it necessary to pre-screen many batches in order to obtain a single one which is well suited for a given application. For example, even a brief exposure of PBMC (peripheral blood mononuclear cells) to a mitogenic serum batch during washing or freezing of these cells will result in a high background in cytokine assays, and toxic/inhibitory serum batches will jeopardize the assay results.  The unique performance of each serum batch necessitates the purchase of large lots to assure consistency of assay conditions as long as possible.  This adds to the inconvenience and cost associated with the use of serum.  Switching to, or using a different batch of serum introduces each time a significant unpredictable variable to the test conditions, making test results difficult to compare. 

For all these reasons, there is considerable pressure from regulatory agencies and from the scientific community to avoid the use of serum and move to consistent, defined substitutes.  However, because serum is naturally rich in a multitude of growth factors and other essentials of cell growth and functionality, it has been challenging to develop a serum substitute for primary cells in general, and for PBMC in particular.  Most serum free media are “rich enough” to permit the growth of robust tumor cells (e.g. hybridomas), but freshly isolated human PBMC require more stringent conditions to maintain their viability and activity in serum-free media. 

Cellular Technology Ltd. (CTL) has been working for years with the US Government, industrial- and academic partners to standardize ELISPOT and other direct ex vivo cytokine assays performed on freshly isolated and cryopreserved PBMC.  These efforts have resulted in CTL’s serum-free media platform. 

CTL’s serum-free platform consists of three product lines that have been carefully designed to cover the three critical steps of working with human PBMC: washing, testing and freezing.  All three previously required the use of serum.

 

Click here for CTL's Serum-Free Media Scientific Resource section

Serum-Free Media Products


CTL Cryo™ Medium Kit

When frozen according to our protocols, PBMC maintain their full functionality in ELISPOT assays. CTL Cryo™ Medium Kit provides all of the necessary components for freezing.

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Wash Media Supplement

CTL’s new serum-free washing media offers a standardized, convenient and cost-efficient alternative.

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Test Media

CTL’s new serum-free testing media consistently provide lower noise and higher signal in such assays that even the “best” serum batches.

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