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Updated: Jan 30, 2026

A Versatile Automated Platform for Micro-scale Cell Stimulation Experiments
Published on: August 6, 2013
A versatile platform for sequential glyco-, phospho-, and proteomics with multi-PTMs integration
Xuefang Dong1,2, Fangfang Xiong1,2, Guangzhu Du1,3
1State Key Laboratory of Phytochemistry and Natural Medicines, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, P. R. China.
Abstract:
Serial multi-omic analysis of proteome, phosphoproteome, and glycoproteome is pivotal for elucidating drug mechanisms, discovering biomarkers, and identifying therapeutic targets. However, simultaneous multi-level post-translational modifications (PTMs) analysis via parallel processing is hampered by laborious, time-consuming procedures and inconsistent reproducibility. We present an integrated Multi-level PTMs-Proteomic Enrichment platform (MuPPE), enabling sequential glycoproteome, phosphoproteome, and proteome analysis from single biological samples. It combines protein aggregation capture with on-bead digestion and tandem enrichment, achieving superior reproducibility (CV 12.3% vs 17.6% conventional methods) while reducing processing time by 87.5% (4 hours vs 32 hours). MuPPE also enhances coverage, identifying more serum glycopeptides and brain phosphopeptides than other platforms. Applied to aging mouse cohorts, the platform uncovers tissue-specific PTMs remodeling and brain barrier dysfunction. For arsenic mechanisms of action, MuPPE reveals drug-induced PTMs crosstalk between glycosylation and phosphorylation-driven pathway regulation. MuPPE offers a transformative tool for advancing multi-omics insights across precision medicine and disease research.
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