Fully automated sample treatment method for high throughput proteome analysis

• 作者: Huiming Yuan Zhongpeng Dai Xiaodan Zhang Baofeng Zhao Hongwei Chu Lihua Zhang Yukui Zhang
• 来源期刊: 中国科学：化学（英文版）
• 年，卷（期）: 2021,（2）
• 所属期刊栏目: ARTICLES
• 页码范围: 313-321
• 页数: 9页
• 语种: 英文

摘要

The bottom-up strategy for proteome analysis typically employs a multistep sample preparation workflow that suffers from being time-consuming and sample loss or contamination caused by the off-line manual operation.Herein,we developed a hollow fibre membrane(HFM)-aided fully automated sample treatment(FAST)method.Due to the confinement effects of HFMs and the immobilized enzymatic reactor,the proteome samples could be denatured,reduced,desalted and digested within 8-20 min via the one-stop service.This method also showed superiority in trace sample analysis.In one and half hours,we could identify about 1,600 protein groups for 500 HeLa cells as the starting materials,1.5-8 times more than those obtained by previously reported methods.Through the on-line combination of FAST with nano-liquid chromatography-electrospray ionization tandem mass spectrometry(nanoLC-ESI-MS/MS),we further established a fully integrated platform for label-free quantification of proteome with high reproducibility and precision.Collectively,FAST presented here represents a major advance in the high throughput sample treatment and quantitative analysis of proteomes.