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

被引：145

作者：

Dou, Maowei ^{[1
]}

Clair, Geremy ^{[2
]}

Tsai, Chia-Feng ^{[2
]}

Xu, Kerui ^{[1
]}

Chrisler, William B. ^{[2
]}

Sontag, Ryan L. ^{[2
]}

Zhao, Rui ^{[1
]}

Moore, Ronald J. ^{[2
]}

Liu, Tao ^{[2
]}

Pasa-Tolic, Ljiljana ^{[1
]}

Smith, Richard D. ^{[2
]}

Shi, Tujin ^{[2
]}

Adkins, Joshua N. ^{[2
]}

Qian, Wei-Jun ^{[2
]}

Kelly, Ryan T. ^{[1
,3
]}

Ansong, Charles ^{[2
]}

Zhu, Ying ^{[1
]}

机构：

[1] Pacific Northwest Natl Lab, Environm Mol Sci Lab, Richland, WA 99354 USA

[2] Pacific Northwest Natl Lab, Biol Sci Div, Richland, WA 99354 USA

[3] Brigham Young Univ, Dept Chem & Biochem, Provo, UT 84604 USA

来源：

ANALYTICAL CHEMISTRY | 2019年 / 91卷 / 20期

关键词：

TANDEM MASS TAGS; LIQUID-CHROMATOGRAPHY; HETEROGENEITY; QUANTIFICATION; SPECTROMETRY; GENE; QUANTITATION; DISCOVERY; PACKAGE; SYSTEM;

D O I：

10.1021/acs.analchem.9b03349

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

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 similar to 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.

引用

页码：13119 / 13127

页数：9

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