Protocol | Published:

Guidelines for the automated evaluation of Elispot assays

Nature Protocols volume 10, pages 10981115 (2015) | Download Citation

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.

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Acknowledgements

C.M.B. and H.S. were supported by a grant from the Wallace Coulter Foundation (Florida, USA). We thank all participants in the Elispot plate reading panel and consensus process. The full list of participants, including several authors of this paper, is given in the Supplementary Note. We also thank A. Szterenfeld for helpful comments on the manuscript.

Author information

Author notes

    • Cedrik M Britten

    Present address: Immuno-Oncology and Combinations DPU, Oncology R&D, GlaxoSmithKline, Stevenage, UK.

Affiliations

  1. ZellNet Consulting, Inc., Fort Lee, New Jersey, USA.

    • Sylvia Janetzki
  2. LBPrice Statistical Consulting Ltd., Karmiel, Israel.

    • Leah Price
  3. BioNTech Diagnostics GmbH, Mainz, Germany.

    • Helene Schroeder
  4. Association for Cancer Immunotherapy (CIMT), Mainz, Germany.

    • Cedrik M Britten
  5. Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands.

    • Marij J P Welters
  6. Department of ImmunoOncology, GlaxoSmith Kline, Collegeville, Pennsylvania, USA.

    • Axel Hoos

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Contributions

S.J., L.P., C.M.B. and M.J.P.W. designed the study. S.J., L.P., H.S., C.M.B., M.J.P.W. and A.H. organized and led the panel. All panelists listed under acknowledgement provided data and feedback leading to the final protocol. S.J. and L.P. compiled and analyzed the data. S.J. wrote the manuscript.

Competing interests

S.J. is founder and president of ZellNet Consulting, Inc. L.P. is the founder and director of LBPrice Statistical Consulting, Ltd. All other authors declare no competing financial interests.

Corresponding author

Correspondence to Sylvia Janetzki.

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    Supplementary Text and Figures

    Supplementary Data, Supplementary Table 1 and Supplementary Note

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DOI

https://doi.org/10.1038/nprot.2015.068

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