U.S. Department of Energy

Pacific Northwest National Laboratory

Facile carrier-assisted targeted mass spectrometric approach for proteomic analysis of low numbers of mammalian cells

TitleFacile carrier-assisted targeted mass spectrometric approach for proteomic analysis of low numbers of mammalian cells
Publication TypeJournal Article
Year of Publication2018
AuthorsShi T., Gaffrey M.J, Fillmore T.L, Nicora C.D, Yi L., Zhang P., Shukla A.K, Wiley H.S, Rodland K.D, Liu T., Smith R.D, Qian W.J
JournalCommun Biol
Abstract

There is an unmet technical challenge for mass spectrometry (MS)-based proteomic analysis of single mammalian cells. Quantitative proteomic analysis of single cells has been previously achieved by antibody-based immunoassays but is limited by the availability of high-quality antibodies. Herein we report a facile targeted MS-based proteomics method, termed cPRISM-SRM (carrier-assisted high-pressure, high-resolution separations with intelligent selection and multiplexing coupled to selected reaction monitoring), for reliable analysis of low numbers of mammalian cells. The method capitalizes on using "carrier protein" to assist processing of low numbers of cells with minimal loss, high-resolution PRISM separation for target peptide enrichment, and sensitive SRM for protein quantification. We have demonstrated that cPRISM-SRM has sufficient sensitivity to quantify proteins expressed at ≥200,000 copies per cell at the single-cell level and ≥3000 copies per cell in 100 mammalian cells. We envision that with further improvement cPRISM-SRM has the potential to move toward targeted MS-based single-cell proteomics.

| Pacific Northwest National Laboratory