Chem. J. Chinese Universities ›› 2021, Vol. 42 ›› Issue (10): 3062.doi: 10.7503/cjcu20210256

• Analytical Chemistry • Previous Articles     Next Articles

Glucose-6-phosphate Functionalized Hydrophilic Magnetic Probe: a Dual-purpose Affinity Material for Effective Separation and Enrichment of Glycopeptides/Phosphopeptides

WANG Baichun, YUAN Yuxin, YAN Yinghua(), DING Chuanfan(), TANG Keqi   

  1. Key Laboratory of Advanced Mass Spectrometry and Molecular Analysis of Zhejiang Province,Institute of Mass Spectrometry,School of Material Science and Chemical Engineering,Ningbo University,Ningbo 315211,China
  • Received:2021-04-16 Online:2021-10-10 Published:2021-10-10
  • Contact: YAN Yinghua,DING Chuanfan E-mail:yanyinghua@nbu.edu.cn;dingchuanfan@nbu.edu.cn

Abstract:

Glucose-6-phosphate(G6P) functionalized hydrophilic magnetic probe Fe3O4@PDA@TiO2@G6P was constructed by the layer-by-layer modification on the surface of the magnetic core Fe3O4. Polydopamine(PDA) not only enhanced the hydrophilicity of the probe, but also acted as the coupling linker for the further grafting of titanium dioxide(TiO2). In addition to serving as the anchor point of G6P, the grafted TiO2 could also effectively enrich phosphopeptides through the metal oxide affinity chromatography technology. The functionalization of G6P gave the nanospheres a highly hydrophilic surface and used the hydrophilic interaction liquid chromatography technology to achieve the goal of capturing glycopeptides. The results showed that this dual-purpose affinity probe had a low detection limit(0.1 fmol/μL), a high selectivity[m(HRP)∶m(BSA)=1∶1000], a good repeatability(ten cycles) and a high loading(300 mg/g) for glycopeptides. Besides, as for phosphopeptides, the obtained G6P functionalized hydrophilic magnetic probe exhibited a low detection limit(0.02 fmol/μL) and a high selectivity[nβ-casein)∶n(BSA)=1∶1000]. At the same time, this dual-purpose affinity material had the ability to simultaneously enrich glycopeptides and phosphopeptides, and 34 glycopeptides and 36 phosphopeptides were identified from human saliva, indicating that it will show great application prospects in proteomics analysis of a variety of post-translational modifications.

Key words: Dual-purpose hydrophilic magnetic probe, Glucose-6-phosphate, Glycopeptide/phosphopeptide, Mass spectrometry detection (Ed.: N, K)

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