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

Overview

Journal Cells

Publisher MDPI

Specialties Biochemistry
Biophysics
Cell Biology
Molecular Biology

Date 2018 Jan 6

PMID 29301355

Citations 2

Authors

Alexander Lehmann

Zoltan Megyesi

Anna Przybyla

Paul V Lehmann

Affiliations

Soon will be listed here.

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.

Citing Articles

Unbiased, High-Throughput Identification of T Cell Epitopes by ELISPOT.

Lehmann P, Roen D, Lehmann A Methods Mol Biol. 2023; 2673:69-88.

PMID: 37258907 DOI: 10.1007/978-1-0716-3239-0_5.

Advances of Single-Cell Protein Analysis.

Liu L, Chen D, Wang J, Chen J Cells. 2020; 9(5).

PMID: 32443882 PMC: 7290353. DOI: 10.3390/cells9051271.

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