Integrated Protein Precipitation–Solid-Phase Extraction Workflow for Deep Top-Down Proteomic Analysis of Human Serum

Journal of Proteome Research 2026

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Authors: Ning Liu, Meng Zhang, Huiwen Qin, Hongyun Zhao, Y C Zhu, Y D Xie, Jiaxiang Liu, Fangjun Wang, Zheyi Liu, Cheng Chang
DOI: 10.1021/acs.jproteome.6c00159
Cited by: 1
References: 61

Abstract

Top-down proteomics of human serum is hindered by extreme dynamic range and cumbersome prefractionation. Here, we establish a fully solution-based protein precipitation–solid-phase extraction (PP–SPE) workflow for deep, low-molecular-weight-biased proteoform profiling. Using systematically optimized combinations of five precipitants and three sorbents, the workflow enabled identification of 4470 proteoforms from 480 proteins with excellent reproducibility ( R > 0.96). Compared with the gel-based PEPPI method, PP–SPE provided substantially greater depth, including >10-fold more

Fields of Study & Subjects

Advanced Proteomics Techniques and Applications Analytical chemistry methods development Advanced Biosensing Techniques and Applications

Publication details

Journal / source
Journal of Proteome Research
Publisher
American Chemical Society (ACS)
Volume
25
Issue
7
Pages
3622–3633
Publication date
3 June 2026
Language
EN