Nanodroplet processing platform for deep and quantitative proteome profiling of 10-100 mammalian cells

作者：Zhu, Ying; Piehowski, Paul D.; Zhao, Rui; Chen, Jing; Shen, Yufeng; Moore, Ronald J.; Shukla, Anil K.; Petyuk, Vladislav A.; Campbell-Thompson, Martha; Mathews, Clayton E.; Smith, Richard D.; Qian, Wei-Jun; Kelly, Ryan T.^*

来源：Nature Communications, 2018, 9(1): 882.

DOI：10.1038/s41467-018-03367-w

• MASS-SPECTROMETRY
• COMPUTATIONAL PLATFORM
• SHOTGUN PROTEOMICS
• DROPLET ARRAY
• RNA-SEQ
• HETEROGENEITY
• DIGESTION
• PROTEINS
• TISSUES
• SAMPLE

Nanoscale or single-cell technologies are critical for biomedical applications. However, current mass spectrometry (MS)-based proteomic approaches require samples comprising a minimum of thousands of cells to provide in-depth profiling. Here, we report the development of a nanoPOTS (nanodroplet processing in one pot for trace samples) platform for small cell population proteomics analysis. NanoPOTS enhances the efficiency and recovery of sample processing by downscaling processing volumes to <200 nL to minimize surface losses. When combined with ultrasensitive liquid chromatography-MS, nanoPOTS allows identification of similar to 1500 to similar to 3000 proteins from similar to 10 to similar to 140 cells, respectively. By incorporating the Match Between Runs algorithm of MaxQuant, >3000 proteins are consistently identified from as few as 10 cells. Furthermore, we demonstrate quantification of similar to 2400 proteins from single human pancreatic islet thin sections from type 1 diabetic and control donors, illustrating the application of nanoPOTS for spatially resolved proteome measurements from clinical tissues.

• 出版日期2018-2-28