The 15-min Subcellular Proteome

This project integrated data-independent acquisition (DIA) with capillary electrophoresis electrospray ionization mass spectrometry (CE-ESI-MS) to enable fast single-cell proteomics. With <15 min of effective separation time in a bottom-up proteomics setting, this technology returned ~1,200 proteins from a single HeLa-cell-equivalent proteome amount. Furthermore, using capillary microsampling to collect the proteome from identified cells in a cleavage-stage embryo of the vertebrate Xenopus laevis (frog) embryo, ~1,400 proteins were identified by measuring ~5 ng of the subcellular proteome content aspirated from live Xenopus embryos. CE-ESI-MS with DIA enhanced proteome detection sensitivity and improved analytical throughput.