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

Charles Bruce Bagwell, Benjamin Hunsberger, Beth Hill, Donald Herbert, Christopher Bray, Thirumahal Selvanantham, Stephen Li, Jose C. Villasboas, Kevin Pavelko, Michael Strausbauch, Adeeb Rahman, Gregory Kelly, Shahab Asgharzadeh, Azucena Gomez-Cabrero, Gregory Behbehani, Hsiaochi Chang, Justin Lyberger, Ruth Montgomery, Yujiao Zhao, Margaret InokumaOfir Goldberger, Greg Stelzer

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)146-160
Number of pages15
JournalCytometry Part B - Clinical Cytometry
Volume98
Issue number2
DOIs
StatePublished - Mar 1 2020

Keywords

  • cytometry automation
  • cytometry standardization
  • kits
  • percentage precision

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

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    Bagwell, C. B., Hunsberger, B., Hill, B., Herbert, D., Bray, C., Selvanantham, T., Li, S., Villasboas, J. C., Pavelko, K., Strausbauch, M., Rahman, A., Kelly, G., Asgharzadeh, S., Gomez-Cabrero, A., Behbehani, G., Chang, H., Lyberger, J., Montgomery, R., Zhao, Y., ... Stelzer, G. (2020). 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. Cytometry Part B - Clinical Cytometry, 98(2), 146-160. https://doi.org/10.1002/cyto.b.21858