Citation: FAN Chao, SONG Zifeng, QIN Weijie, CAI Yun, QIAN Xiaohong. A novel immobilized trypsin on hydrophilic polymer modified silica beads for proteome characterization[J]. Chinese Journal of Chromatography, ;2013, 31(5): 423-428. doi: 10.3724/SP.J.1123.2013.01007 shu

A novel immobilized trypsin on hydrophilic polymer modified silica beads for proteome characterization

FAN Chao ^1,2 ,
SONG Zifeng ² ,
QIN Weijie ^2,, ,
CAI Yun ^2,, ,
QIAN Xiaohong ²

1.
1. Graduate Department of Anhui Medical University, Hefei 230032, China;

Corresponding author: QIN Weijie, CAI Yun,
Received Date: 5 January 2013
Available Online: 7 February 2013
Fund Project: 国家重大科学计划项目(2013CB911204) (2013CB911204)国家重大科学仪器设备开发专项项目(2011YQ09000504) (2011YQ09000504)国家自然科学基金项目(21275005,21235001). (21275005,21235001)

A new type of immobilized trypsin was prepared using hydrophilic polymer modified silica microparticles (HPMSM) synthesized by atom transfer radical polymerization (ATRP) as the substrate material. ATRP modification led to densely packed hydrophilic polymer chains grafted on the microparticles surface which resulted in largely increased trypsin loading amount and minimized the nonspecific adsorption of proteins and peptides. Therefore, ultra-fast and highly efficient protein digestion was achieved with minimized sample loss. For standard protein bovine serum albumin (BSA), 1 min digestion led to the identification of 93 peptides, which covered 91% amino acid sequence of the protein. This immobilized trypsin was further successfully applied to the digestion of complex protein samples from yeast and 666 proteins were identified after 3 min digestion, which exceeded the number of identified proteins after 12 h solution digestion.
- immobilized trypsin,
- proteomics,
- atom transfer radical polymerization (ATRP)
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