CIMAC-CIDC CyTOF harmonization
| Autor: | Salah Eddine Bentebibel, Nicholas F. Fernandez, Sean C. Bendall, Beatriz Sanchez Espiridion, Emily M. Thrash, Karen A. Millerchip, Natalia Sigal, Bita Sahaf, Adeeb Rahman, Chantale Bernatchez, Sacha Gnjatic, Ignacio I. Wistuba, Mina Pichavant, Emily M. McWilliams, Cara Haymaker, Diane Marie Del Valle, Caroline Duault, Holden T. Maecker, Melanie Davila |
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| Rok vydání: | 2020 |
| Předmět: | |
| Zdroj: | Journal of Clinical Oncology. 38:e15242-e15242 |
| ISSN: | 1527-7755 0732-183X |
| Popis: | e15242 Background: The Cancer Immune Monitoring and Analysis Centers – Cancer Immunology Data Commons (CIMAC-CIDC) network is a National Cancer Institute-funded initiative to identify biomarkers of mechanisms and response to cancer immunotherapy clinical trials, using state-of-the-art assay technologies. A primary platform for CIMAC-CIDC biomarker studies is CyTOF mass cytometry, which is performed at all four CIMAC laboratories. Methods: To test the ability to generate comparable data across labs, a cross-site harmonization effort was undertaken. We first harmonized SOPs between centers. Because of a new acquisition protocol introduced by the vendor (Fluidigm), we also tested this protocol across sites before finalizing the harmonized SOP. We then performed a cross-site assay harmonization experiment, using 5 shared cryopreserved PBMC samples and one lyophilized control cell preparation, along with a shared lyophilized antibody cocktail consisting of 14 markers, as validated in the HIPC consortium, plus CD45. These reagents and samples were distributed to the four sites, and FCS files were centrally analyzed by both manual gating and automated methods (Astrolabe). Results: Average CVs across sites for each cell population were reported and compared to a previous multisite CyTOF study. Once a cell recovery issue at two sites was resolved, this experiment resulted in inter-site reproducibility of under 20% CV for most cell subsets, very similar to the previous study. Conclusions: These results emphasize the ability to reproduce CyTOF across sites, and also highlights procedures, such as use of spike-in control samples, useful for tracking variability in this assay. |
| Databáze: | OpenAIRE |
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