Abstract
A novel application of ICP-MS to cell biology is presented. This work details a method of multiplex cellular antigen determination using ICP-MS-linked metal-tagged immunophenotyping. Expression of intracellular oncogenic kinase BCR/Abl, myeloid cell surface antigen CD33, human stem cell factor receptor c -Kit and integrin receptor VLA-4 were investigated using human leukemic cell lines. Antigens to which specific antibodies are available and are distinguishably tagged can be determined simultaneously, or multiplexed. Four commercially available tags (Au, Sm, Eu, and Tb) conjugated to secondary antibodies enable a 4-plex assay assuming that the primary antibodies are not cross-reactive. Depending on the abundance of antigens it was possible to detect as little as 1000 cells in a mixed cell population. Results obtained by ICP-MS were compared with data from conventional flow cytometry. ICP-MS as an analytical detector possesses several advantages that enhance the performance of immunoassays, which are discussed in detail. Although multiplexing using metal-conjugated reagents is in a very early stage of research and feasibility studies, it is already apparent that more than four could be accurately detected simultaneously using the ICP-MS instrument.
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