Flow Cytometry Method Validation Protocols

Abstract

AbstractAnalytical method validation provides a means to ensure that data are credible and reproducible. This article will provide a brief introduction to analytical method validation as applied to cellular analysis by flow cytometry, along with practical procedures for four different types of validation. The first, Basic Protocol 1 (the limited validation protocol), is recommended for research and non‐regulated laboratories. Next, Basic Protocol 2) presents a reasonable, fit‐for‐purpose validation approach appropriate for biopharma and research settings. Basic Protocol 3 addresses the type of validation performed in clinical laboratories for moderate‐risk tests developed in house. Finally, Basic Protocol 4 describes the process that should be applied whenever a method is being transferred from one facility to another. All four validation plans follow the fit‐for‐purpose validation approach, in which the validation parameters are selected based on the intended use of the assay. These validation protocols represent the minimal requirement and may not be applicable for every intended use such as high‐risk clinical assays or data to be used as a primary endpoint in a clinical trial. The recommendations presented here are consistent with the white papers published by the American Association of Pharmaceutical Scientists and the International Clinical Cytometry Society, as well as with Clinical Laboratory Standards Institute Guideline H62: Validation of Assays Performed by Flow Cytometry (CLSI, 2021). © 2023 Wiley Periodicals LLC.Basic Protocol 1: Limited validationBasic Protocol 2: Fit‐for‐purpose validation for biopharma and research settingsBasic Protocol 3: Validation for moderate clinical risk laboratory developed testsBasic Protocol 4: Transfer validationThis article was corrected on 25 January 2024. See the end of the full text for details.