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Cells 2018, 7(1), 3; doi:10.3390/cells7010003

Reagent Tracker Dyes Permit Quality Control for Verifying Plating Accuracy in ELISPOT Tests

1
Research and Development Department, CTL, Shaker Heights, OH 44122, USA
2
Department of Cancer Immunology, Chair of Medical Biotechnology, Poznan University of Medical Sciences, 61-701 Poznan, Poland
*
Author to whom correspondence should be addressed.
Received: 4 December 2017 / Revised: 25 December 2017 / Accepted: 29 December 2017 / Published: 3 January 2018
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Abstract

ELISPOT assays enable the detection of the frequency of antigen-specific T cells in the blood by measuring the secretion of cytokines, or combinations of cytokines, in response to antigenic challenges of a defined population of PBMC. As such, these assays are suited to establish the magnitude and quality of T cell immunity in infectious, allergic, autoimmune and transplant settings, as well as for measurements of anti-tumor immunity. The simplicity, robustness, cost-effectiveness and scalability of ELISPOT renders it suitable for regulated immune monitoring. In response to the regulatory requirements of clinical and pre-clinical immune monitoring trials, tamper-proof audit trails have been introduced to all steps of ELISPOT analysis: from capturing the raw images of assay wells and counting of spots, to all subsequent quality control steps involved in count verification. A major shortcoming of ELISPOT and other related cellular assays is presently the lack of audit trails for the wet laboratory part of the assay, in particular, the assurance that no pipetting errors have occurred during the plating of antigens and cells. Here, we introduce a dye-based reagent tracking platform that fills this gap, thereby increasing the transparency and documentation of ELISPOT test results. View Full-Text
Keywords: ImmunoSpot®; antigen screening; determinant mapping; CD4 cells; CD8 cells; T cells; RT dyes; audit trails for ELISPOT; regulated ELISPOT ImmunoSpot®; antigen screening; determinant mapping; CD4 cells; CD8 cells; T cells; RT dyes; audit trails for ELISPOT; regulated ELISPOT
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Lehmann, A.; Megyesi, Z.; Przybyla, A.; Lehmann, P.V. Reagent Tracker Dyes Permit Quality Control for Verifying Plating Accuracy in ELISPOT Tests. Cells 2018, 7, 3.

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