title: Mass cytometry panel optimization through the designed distribution of signal interference.
abstract: Mass cytometry is capable of measuring more than 40 distinct proteins on individual cells making it a promising technology for innovating biomarker discovery. However, in order for this potential to be fully realized, best practices in panel design need to be further defined in order to achieve consistency and reproducibility in data analysis. Of particular importance are controls that reveal, and panel design principles that mitigate the effects of signal interference or overlap. We observed a disparity between the staining profiles of two noncompeting anti- integrin β7 mAbs and hypothesized that signal interference was responsible. A mass-minus-one (MMO) control was applied and demonstrated that signal overlap caused the perceived interclonal discrepancy in β7 expression. Panel redesign in consideration of mass-cytometry specific interference dynamics dramatically improved concordance between both mAbs by redistributing background signals caused by overlap. These studies visualize how signal overlap can complicate mass cytometry data interpretation and demonstrate how the rational distribution of interference can greatly improve panel design and data quality. © 2016 International Society for Advancement of Cytometry.
authors.affiliation: OMNI Biomarkers Department, Development Sciences, Genentech, South San Francisco, California, 94080.
authors.forename: Chikara
authors.initials: C
authors.lastname: Takahashi
authors.affiliation: OMNI Biomarkers Department, Development Sciences, Genentech, South San Francisco, California, 94080.
authors.forename: Amelia
authors.initials: A
authors.lastname: Au-Yeung
authors.affiliation: OMNI Biomarkers Department, Development Sciences, Genentech, South San Francisco, California, 94080.
authors.forename: Franklin
authors.initials: F
authors.lastname: Fuh
authors.affiliation: OMNI Biomarkers Department, Development Sciences, Genentech, South San Francisco, California, 94080.
authors.forename: Teresa
authors.initials: T
authors.lastname: Ramirez-Montagut
authors.affiliation: Bioinformatics Department, Genentech, South San Francisco, California, 94080.
authors.forename: Chris
authors.initials: C
authors.lastname: Bolen
authors.affiliation: OMNI Biomarkers Department, Development Sciences, Genentech, South San Francisco, California, 94080.
authors.forename: William
authors.initials: W
authors.lastname: Mathews
authors.affiliation: OMNI Biomarkers Department, Development Sciences, Genentech, South San Francisco, California, 94080.
authors.forename: William E
authors.initials: WE
authors.lastname: O'Gorman
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods*; D005786:Gene Expression Regulation; D006801:Humans; D039641:Integrin beta Chains / Q000096:biosynthesis* / Q000276:immunology; D007963:Leukocytes, Mononuclear / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D000911:Antibodies, Monoclonal; D039641:Integrin beta Chains; C091368:integrin beta7
keywords: CyTOF; mass cytometry; panel optimization; signal interference