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- W2462236718 abstract "Background Validated assays are essential to generate data with defined specificity, consistency, and reliability. Although the process of validation is required for applying immunoassays in the context of clinical studies, reports on systematic validation of in vitro T cell assays are scarce so far. We recently validated our HLA‐peptide multimer staining assay in a systematic manner so as to qualify the method for monitoring antigen‐specific T cell responses after immunotherapy. Methods Parameters of the assay, specificity, precision, linearity, sensitivity, and robustness were assessed systematically. Experiments were designed to address specifically each parameter and are detailed. Results Nonspecific multimer staining was below the acceptance limit of 0.02% multimer (+) CD8 (+) cells. The assay showed acceptable precision in all dimensions it was repeated (CV < 10%) and also demonstrated a linear detection ( R 2 > 0.99) of antigen specific cells. Conclusions We succeeded in validating the HLA‐multimer staining assay in a systematic manner. Additionally, we propose a technical framework and recommendations that can be applied for validating other T cell assessment methods. © 2016 International Clinical Cytometry Society" @default.
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- W2462236718 date "2016-07-22" @default.
- W2462236718 modified "2023-09-27" @default.
- W2462236718 title "Validation of Immunomonitoring Methods for Application in Clinical Studies: The HLA-Peptide Multimer Staining Assay" @default.
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- W2462236718 doi "https://doi.org/10.1002/cyto.b.21397" @default.
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