The complex nature of the immune system requires deep interrogation at the single-cell level. Mass cytometry, which uses CyTOF® technology, utilizes antibodies tagged with monoisotopic metals, resulting in discrete signals that enable highly multiparametric characterization on a single-cell level. CyTOF can currently enable cellular phenotyping of over 50 single-cell parameters simultaneously, including phenotypic and functional markers, thereby significantly increasing the ability to comprehensively evaluate immune responses.
The Maxpar® Direct™Immune Profiling Assay™is a pre-titrated, dried-down, 30-marker antibody cocktail for immune profiling of human whole blood and peripheral blood mononuclear cells (PBMC) by CyTOF (Figure 1). Using Maxpar Pathsetter™software, stained samples are automatically resolved into 37 immune populations. This assay was designed with customization in mind, with 18-plus open channels available for panel expansion.
This study highlights the use of the Maxpar Direct T Cell Activation Expansion Panel and live-cell barcoding to add 15 parameters to the backbone Maxpar Direct panel. The array of key surface and intracellular functional parameters in this expanded panel enables comprehensive analysis of immune cell activation and antigen-specific recall responses. Such phenomena are hallmarks for research on infection, vaccine development, and immunotherapy.