Platinum-conjugated antibodies for application in mass cytometry.
Mass cytometry has overcome limitations of fluorescent single cell cytometry by allowing for the measurement of up to currently ∼40 different parameters on a single cell level. However, the cellular proteome comprises many more potential analytes, and current mass cytometry instrumentation allows for theoretically up to 121 different mass detection channels. The labeling of specific probes with appropriate metal ions is a significant hurdle for exploiting more of mass cytometry's analytical capacity. To this end, we here describe the labeling of antibody with natural abundance or isotopically purified platinum as formulated in cisplatin and circumventing the use of chelator-loaded polymers. We confirm the utility of cisplatin-antibody-conjugates for surface, intracellular, and phosphoepitope-specific immunophenotyping, as well as for application in cell surface CD45-based barcoding. Cisplatin-labeling of antibody increases the analytical capacity of the CyTOF(®) platform by two channels based on available reagents, and has the potential to add a total of six channels for detection of specific probes, thus helping to better extend the analytical mass range of mass cytometers.
The Human Immune Monitoring Center (HIMC), Institute for Immunity, Transplantation and Infection, Stanford University School of Medicine, Stanford, California, 94305.
Henrik E
HE
Mei
The Human Immune Monitoring Center (HIMC), Institute for Immunity, Transplantation and Infection, Stanford University School of Medicine, Stanford, California, 94305.
Michael D
MD
Leipold
The Human Immune Monitoring Center (HIMC), Institute for Immunity, Transplantation and Infection, Stanford University School of Medicine, Stanford, California, 94305.
Holden T
HT
Maecker
D000906:Antibodies / Q000737:chemistry*; D002945:Cisplatin / Q000737:chemistry*; D005434:Flow Cytometry / Q000379:methods*; D015870:Gene Expression; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry; D016130:Immunophenotyping / Q000379:methods*; D028581:Lanthanoid Series Elements / Q000032:analysis; D017493:Leukocyte Common Antigens / Q000235:genetics / Q000276:immunology; D008213:Lymphocyte Activation; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D009000:Monocytes / Q000166:cytology / Q000276:immunology; D059010:Single-Cell Analysis / Q000379:methods; D013194:Staining and Labeling / Q000379:methods; D013601:T-Lymphocytes / Q000145:classification / Q000166:cytology* / Q000276:immunology
D000906:Antibodies; D018796:Immunoconjugates; D028581:Lanthanoid Series Elements; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human; D002945:Cisplatin
CD45; CyTOF; antibody conjugation; barcoding; biomarker; cisplatin; immunomonitoring; immunophenotyping; mass cytometry; mass-tagged antibody; metal reporter; platinum