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Nat Protoc. 2015 Jul;10(7):1098-115. doi: 10.1038/nprot.2015.068. Epub 2015 Jun 25.

Guidelines for the automated evaluation of Elispot assays.

Author information

1
ZellNet Consulting, Inc., Fort Lee, New Jersey, USA.
2
LBPrice Statistical Consulting Ltd., Karmiel, Israel.
3
BioNTech Diagnostics GmbH, Mainz, Germany.
4
Association for Cancer Immunotherapy (CIMT), Mainz, Germany.
5
Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands.
6
Department of ImmunoOncology, GlaxoSmith Kline, Collegeville, Pennsylvania, USA.

Abstract

The presented protocol for Elispot plate evaluation summarizes how to implement the recommendations developed following the establishment of a large-scale international Elispot plate-reading panel and subsequent multistep consensus-finding process. The panel involved >100 scientists from various immunological backgrounds. The protocol includes the description and justification of steps for setting reading parameters to obtain accurate, reliable and precise automated analysis results of Elispot plates. Further, necessary adjustments for out-of-specification situations are described and examples are provided. The plate analysis, including parameter adjustments, auditing of results and necessary annotations, should be achievable within a time range of 10-30 min per plate. Adoption of these guidelines should enable a further reduction in assay variability and an increase in the reliability and comparability of results obtained by Elispot. These guidelines conclude the ongoing harmonization efforts for the enzymatic Elispot assay.

PMID:
26110715
DOI:
10.1038/nprot.2015.068
[Indexed for MEDLINE]

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