Streamlined single-cell proteomics by an integrated microfluidic chip and data-independent acquisition mass spectrometry

被引：144

作者：

Gebreyesus, Sofani Tafesse ^{[1
,2
,3
]}

Siyal, Asad Ali ^{[1
,4
,5
]}

Kitata, Reta Birhanu ^{[1
]}

Chen, Eric Sheng-Wen ^{[1
]}

Enkhbayar, Bayarmaa ^{[4
,6
]}

Angata, Takashi ^{[6
]}

Lin, Kuo-I ^{[7
]}

Chen, Yu-Ju ^{[1
,3
,4
,8
,9
]}

Tu, Hsiung-Lin ^{[1
,2
,4
,8
,9
]}

机构：

[1] Acad Sinica, Inst Chem, Taipei 11529, Taiwan

[2] Acad Sinica, Nano Sci & Technol Program, Taiwan Int Grad Program, Taipei 11529, Taiwan

[3] Natl Taiwan Univ, Dept Chem, Taipei 10617, Taiwan

[4] Acad Sinica, Taiwan Int Grad Program, Chem Biol & Mol Biophys Program, Taipei 11529, Taiwan

[5] Natl Tsing Hua Univ, Dept Chem, Hsinchu 30013, Taiwan

[6] Acad Sinica, Inst Biol Chem, Taipei 11529, Taiwan

[7] Acad Sinica, Genom Res Ctr, Taipei 11529, Taiwan

[8] Acad Sinica, Genome & Syst Biol Degree Program, Taipei 10617, Taiwan

[9] Natl Taiwan Univ, Taipei 10617, Taiwan

来源：

NATURE COMMUNICATIONS | 2022年 / 13卷 / 01期

关键词：

LABEL-FREE; FUTURE;

D O I：

10.1038/s41467-021-27778-4

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Single-cell proteomics can reveal cellular phenotypic heterogeneity and cell-specific functional networks underlying biological processes. Here, we present a streamlined workflow combining microfluidic chips for all-in-one proteomic sample preparation and data-independent acquisition (DIA) mass spectrometry (MS) for proteomic analysis down to the single-cell level. The proteomics chips enable multiplexed and automated cell isolation/counting/imaging and sample processing in a single device. Combining chip-based sample handling with DIA-MS using project-specific mass spectral libraries, we profile on average similar to 1,500 protein groups across 20 single mammalian cells. Applying the chip-DIA workflow to profile the proteomes of adherent and non-adherent malignant cells, we cover a dynamic range of 5 orders of magnitude with good reproducibility and <16% missing values between runs. Taken together, the chip-DIA workflow offers all-in-one cell characterization, analytical sensitivity and robustness, and the option to add additional functionalities in the future, thus providing a basis for advanced single-cell proteomics applications.

引用

页数：13

共 45 条

[1] Mass-spectrometric exploration of proteome structure and function [J].