Single Cell Proteomics Using Multiplexed Isobaric Labeling for Mass Spectrometric Analysis.

Single cell proteomics is an emerging field of bioanalysis allowing one to capture proteome profiles of isolated single cells, which is expected to yield additional biological information in comparison with bulk cell analysis. Mass spectrometry-based methods provide unbiased analysis of detectable proteins limited only by technical parameters, such as sensitivity, which necessitates the development of best-practice workflows. Here, we describe the entire experimental design of single cell proteome analysis, exemplified by cultured A549 lung adenocarcinoma cells treated with an anti-cancer drug (methotrexate) and utilizing tandem mass tag (TMTpro™) labeling strategy for mass spectrometric data acquisition.