TY - JOUR
T1 - Multi-site reproducibility of a human immunophenotyping assay in whole blood and peripheral blood mononuclear cells preparations using CyTOF technology coupled with Maxpar Pathsetter, an automated data analysis system
AU - Bagwell, Charles Bruce
AU - Hunsberger, Benjamin
AU - Hill, Beth
AU - Herbert, Donald
AU - Bray, Christopher
AU - Selvanantham, Thirumahal
AU - Li, Stephen
AU - Villasboas, Jose C.
AU - Pavelko, Kevin
AU - Strausbauch, Michael
AU - Rahman, Adeeb
AU - Kelly, Gregory
AU - Asgharzadeh, Shahab
AU - Gomez-Cabrero, Azucena
AU - Behbehani, Gregory
AU - Chang, Hsiaochi
AU - Lyberger, Justin
AU - Montgomery, Ruth
AU - Zhao, Yujiao
AU - Inokuma, Margaret
AU - Goldberger, Ofir
AU - Stelzer, Greg
N1 - Publisher Copyright:
© 2019 International Clinical Cytometry Society
PY - 2020/3/1
Y1 - 2020/3/1
N2 - High-dimensional mass cytometry data potentially enable a comprehensive characterization of immune cells. In order to positively affect clinical trials and translational clinical research, this advanced technology needs to demonstrate a high reproducibility of results across multiple sites for both peripheral blood mononuclear cells (PBMC) and whole blood preparations. A dry 30-marker broad immunophenotyping panel and customized automated analysis software were recently engineered and are commercially available as the Fluidigm® Maxpar® Direct™ Immune Profiling Assay™. In this study, seven sites received whole blood and six sites received PBMC samples from single donors over a 2-week interval. Each site labeled replicate samples and acquired data on Helios™ instruments using an assay-specific acquisition template. All acquired sample files were then automatically analyzed by Maxpar Pathsetter™ software. A cleanup step eliminated debris, dead cells, aggregates, and normalization beads. The second step automatically enumerated 37 immune cell populations and performed label intensity assessments on all 30 markers. The inter-site reproducibility of the 37 quantified cell populations had consistent population frequencies, with an average %CV of 14.4% for whole blood and 17.7% for PBMC. The dry reagent coupled with automated data analysis is not only convenient but also provides a high degree of reproducibility within and among multiple test sites resulting in a comprehensive yet practical solution for deep immune phenotyping.
AB - High-dimensional mass cytometry data potentially enable a comprehensive characterization of immune cells. In order to positively affect clinical trials and translational clinical research, this advanced technology needs to demonstrate a high reproducibility of results across multiple sites for both peripheral blood mononuclear cells (PBMC) and whole blood preparations. A dry 30-marker broad immunophenotyping panel and customized automated analysis software were recently engineered and are commercially available as the Fluidigm® Maxpar® Direct™ Immune Profiling Assay™. In this study, seven sites received whole blood and six sites received PBMC samples from single donors over a 2-week interval. Each site labeled replicate samples and acquired data on Helios™ instruments using an assay-specific acquisition template. All acquired sample files were then automatically analyzed by Maxpar Pathsetter™ software. A cleanup step eliminated debris, dead cells, aggregates, and normalization beads. The second step automatically enumerated 37 immune cell populations and performed label intensity assessments on all 30 markers. The inter-site reproducibility of the 37 quantified cell populations had consistent population frequencies, with an average %CV of 14.4% for whole blood and 17.7% for PBMC. The dry reagent coupled with automated data analysis is not only convenient but also provides a high degree of reproducibility within and among multiple test sites resulting in a comprehensive yet practical solution for deep immune phenotyping.
KW - cytometry automation
KW - cytometry standardization
KW - kits
KW - percentage precision
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U2 - 10.1002/cyto.b.21858
DO - 10.1002/cyto.b.21858
M3 - Article
C2 - 31758746
AN - SCOPUS:85075422474
SN - 1552-4949
VL - 98
SP - 146
EP - 160
JO - Cytometry Part B - Clinical Cytometry
JF - Cytometry Part B - Clinical Cytometry
IS - 2
ER -