One-Shot Single-Cell Proteome and Metabolome Analysis Strategy for the Same Single Cell

Characterizing the profiles of proteome and metabolome at the single-cell level is of great significance in single-cell multiomic studies. Herein, we proposed a novel strategy called one-shot single-cell proteome and metabolome analysis (scPMA) to acquire the proteome and metabolome information in a...

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Published in:Analytical chemistry (Washington) 2024-04, Vol.96 (14), p.5499-5508
Main Authors: Wu, Jie, Xu, Qin-Qin, Jiang, Yi-Rong, Chen, Jian-Bo, Ying, Wei-Xin, Fan, Qian-Xi, Wang, Hui-Feng, Wang, Yu, Shi, Shao-Wen, Pan, Jian-Zhang, Fang, Qun
Format: Article
Language:English
Subjects:
Anticancer properties
Antitumor activity
Antitumour agents
Chromatography, Liquid
Doxorubicin
HeLa Cells
Humans
Injection
Metabolites
Metabolome
Peptides
Proteome - metabolism
Proteomes
Tandem Mass Spectrometry
Tumor cells
Workflow
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Summary:Characterizing the profiles of proteome and metabolome at the single-cell level is of great significance in single-cell multiomic studies. Herein, we proposed a novel strategy called one-shot single-cell proteome and metabolome analysis (scPMA) to acquire the proteome and metabolome information in a single-cell individual in one injection of LC-MS/MS analysis. Based on the scPMA strategy, a total workflow was developed to achieve the single-cell capture, nanoliter-scale sample pretreatment, one-shot LC injection and separation of the enzyme-digested peptides and metabolites, and dual-zone MS/MS detection for proteome and metabolome profiling. Benefiting from the scPMA strategy, we realized dual-omic analysis of single tumor cells, including A549, HeLa, and HepG2 cells with 816, 578, and 293 protein groups and 72, 91, and 148 metabolites quantified on average. A single-cell perspective experiment for investigating the doxorubicin-induced antitumor effects in both the proteome and metabolome aspects was also performed.
ISSN:0003-2700
1520-6882
1520-6882
DOI:10.1021/acs.analchem.3c05659