title: Guidelines for the automated evaluation of Elispot assays.
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.
authors.affiliation: ZellNet Consulting, Inc., Fort Lee, New Jersey, USA.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Janetzki
authors.affiliation: LBPrice Statistical Consulting Ltd., Karmiel, Israel.
authors.forename: Leah
authors.initials: L
authors.lastname: Price
authors.affiliation: BioNTech Diagnostics GmbH, Mainz, Germany.
authors.forename: Helene
authors.initials: H
authors.lastname: Schroeder
authors.affiliation: Association for Cancer Immunotherapy (CIMT), Mainz, Germany.
authors.forename: Cedrik M
authors.initials: CM
authors.lastname: Britten
authors.affiliation: Department of Clinical Oncology, Leiden University Medical Center, Leiden, The Netherlands.
authors.forename: Marij J P
authors.initials: MJ
authors.lastname: Welters
authors.affiliation: Department of ImmunoOncology, GlaxoSmith Kline, Collegeville, Pennsylvania, USA.
authors.forename: Axel
authors.initials: A
authors.lastname: Hoos
mesh_terms: D057205:Automation, Laboratory; D058501:Enzyme-Linked Immunospot Assay / Q000379:methods*; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis; D007963:Leukocytes, Mononuclear / Q000276:immunology; D015203:Reproducibility of Results; D059010:Single-Cell Analysis / Q000379:methods
chemical_list: D007371:Interferon-gamma