Single cell proteome profiling using highly sensitive LC-MS system and In-capillary sample preparation method

摘要

LC/MS-based proteomics has become a fairly mature technology, and often is applied for practical evaluations in both basic and clinical sciences. An issue is that the obtained data are ensemble averages of many cells so that the average results loose information on cellular heterogeneity. Single cell analyses are becoming more common place with other measurement areas such as genomics and transcriptomics based on signal amplification.1-3 These results demonstrate that individual cells have unique characters and should be individually analyzed in some cases. It remains difficult to achieve single cell proteome analysis, especially as one cannot comprehensively amplify MS signals of whole digested peptides. Here we optimize each procedure from sample preparation to LC-MS analysis to minimize loses and optimize signals. Overall, we detect hundreds of proteins from minuscule amount of mammalian cell lysates or single Aplysia Californica neurons. Our ability to perform single cell proteome analysis appears achievable in the near future. References: [1] T. Kalisky and S. Quake, Nature methods (2011), [2]F. Tang et al., Nature methods (2009), [3]R. Zenobi, Science (2013)