American Chemical Society
ac9b03349_si_005.xlsx (6.51 MB)

High-Throughput Single Cell Proteomics Enabled by Multiplex Isobaric Labeling in a Nanodroplet Sample Preparation Platform

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posted on 2019-09-24, 20:13 authored by Maowei Dou, Geremy Clair, Chia-Feng Tsai, Kerui Xu, William B. Chrisler, Ryan L. Sontag, Rui Zhao, Ronald J. Moore, Tao Liu, Ljiljana Pasa-Tolic, Richard D. Smith, Tujin Shi, Joshua N. Adkins, Wei-Jun Qian, Ryan T. Kelly, Charles Ansong, Ying Zhu
Effective extension of mass spectrometry-based proteomics to single cells remains challenging. Herein we combined microfluidic nanodroplet technology with tandem mass tag (TMT) isobaric labeling to significantly improve analysis throughput and proteome coverage for single mammalian cells. Isobaric labeling facilitated multiplex analysis of single cell-sized protein quantities to a depth of ∼1 600 proteins with a median CV of 10.9% and correlation coefficient of 0.98. To demonstrate in-depth high throughput single cell analysis, the platform was applied to measure protein expression in 72 single cells from three murine cell populations (epithelial, immune, and endothelial cells) in <2 days instrument time with over 2 300 proteins identified. Principal component analysis grouped the single cells into three distinct populations based on protein expression with each population characterized by well-known cell-type specific markers. Our platform enables high throughput and unbiased characterization of single cell heterogeneity at the proteome level.