Hollow silica bubble based immobilized trypsin for highly efficient proteome digestion and buoyant separation

doi:10.1039/c6ra12599a

	Hollow silica bubble based immobilized trypsin for highly efficient proteome digestion and buoyant separation
	Huang, Junjie 3; Zhang, Yukui 1; Zhang, Yangjun 3; Qian, Xiaohong 3; Jiao, Fenglong 2,3; Zhai, Rui 3
刊名	RSC ADVANCES
	2016
卷号	6 期号:87 页码:84113-84118
ISSN号	2046-2069
DOI	10.1039/c6ra12599a
文献子类	Article
英文摘要	Tryptic digestion before identification and quantification by mass spectrometry is an indispensable process for most proteomics studies. Conventional in-solution digestion process always suffers from incomplete digestion and is time-consuming and expensive. Development of a novel proteolytic digestion method with fast, high efficiency and reusability of the enzyme is still an urgent task. In this study, new hollow silica bubbles-based immobilized trypsin (trypsin@SiB) was developed via brushes structured glycidyl methacrylate (GMA) grafted on the silica bubbles (SiB) by the reversible addition-fragmentation chain transfer polymerization (RAFT) technique followed by trypsin's immobilization on the microsphere. The new immobilized trypsin not only showed high efficiency and stability but also reusability. Due to the low density (0.6 g cm(-3)) of SiB, it can float over the solution for several minutes so that the immobilized trypsin can be separated and recovered from the system easily. Highly efficient digestion of BSA was achieved with this trypsin@SiB within 1 min and the obtained sequence coverage (92%) was better than that from conventional in-solution digestion for more than 12 h (76%). To further confirm the efficiency of trypsin@SiB for complex proteomic analysis, the protein extracted from human urine was analyzed as a real sample. Within 10 min digestion, 510 protein groups were identified with the LC-MS/MS analysis and database searching, whereas the number of identified proteins after 12 h in-solution digestion was 493 with the same identification conditions. The successful application of trypsin@SiB demonstrated its potential as a high efficient digestion method for future proteomics analysis.
WOS关键词	HYBRID MAGNETIC NANOPARTICLES ; ENZYME IMMOBILIZATION ; GRAPHENE OXIDE ; RADICAL POLYMERIZATION ; PROTEOLYTIC DIGESTION ; ENRICHMENT ; MS ; IDENTIFICATION ; GLYCOPEPTIDES ; NANOTUBES
WOS研究方向	Chemistry
语种	英语
出版者	ROYAL SOC CHEMISTRY
WOS记录号	WOS:000384129300070
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/169866]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
通讯作者	Jiao, Fenglong
作者单位	1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Dalian 116011, Peoples R China 2.Beijing Inst Technol, Sch Life Sci & Technol, Beijing 100081, Peoples R China 3.Beijing Inst Radiat Med, Natl Ctr Prot Sci Beijing, State Key Lab Prote, Beijing 102206, Peoples R China
推荐引用方式 GB/T 7714	Huang, Junjie,Zhang, Yukui,Zhang, Yangjun,et al. Hollow silica bubble based immobilized trypsin for highly efficient proteome digestion and buoyant separation[J]. RSC ADVANCES,2016,6(87):84113-84118.
APA	Huang, Junjie,Zhang, Yukui,Zhang, Yangjun,Qian, Xiaohong,Jiao, Fenglong,&Zhai, Rui.(2016).Hollow silica bubble based immobilized trypsin for highly efficient proteome digestion and buoyant separation.RSC ADVANCES,6(87),84113-84118.
MLA	Huang, Junjie,et al."Hollow silica bubble based immobilized trypsin for highly efficient proteome digestion and buoyant separation".RSC ADVANCES 6.87(2016):84113-84118.