Enzymatic Reactor with Trypsin Immobilized on Graphene Oxide Modified Polymer Microspheres To Achieve Automated Proteome Quantification

doi:10.1021/acs.analchem.7b00682

	Enzymatic Reactor with Trypsin Immobilized on Graphene Oxide Modified Polymer Microspheres To Achieve Automated Proteome Quantification
	Yuan, Huiming 2; Zhang, Shen 1,2; Zhao, Baofeng 2; Weng, Yejing 1,2; Zhu, Xudong 1,2; Li, Senwu 1,2; Zhang, Lihua 2; Zhang, Yukui 2
刊名	ANALYTICAL CHEMISTRY
	2017-06-20
卷号	89 页码:6324-6329
DOI	10.1021/acs.analchem.7b00682
英文摘要	Protein digestion and isotope, labeling are two critical steps in proteome quantification. However, the conventional in-solution protocol unavoidably suffers from disadvantages such as time-consuming, low labeling efficiency, and tedious off-line manual operation, which might affect the quantification accuracy, reproducibility, and throughput. To address these problems, we developed a fully automated proteome quantification platform, in which an ultraperformance immobilized microreactor (upIMER) with graphene-oxide-modified polymer micro spheres as the matrix was developed, to achieve not only the simultaneous protein digestion and O-18 labeling, but also the online integration, with nano-high-pressure liquid chromatography electrospray ionization-tandem mass spectrometry (nanoHPLC ESI-MS/MS). Compared to the conventional off-line protocols, such a platform exhibits obviously improved digestion and O-18 labeling efficiency (only 8% peptides with missed cleavage sites, 99% labeling efficiency, and 2.5 min reaction time), leading to the increased quantification coverage, accuracy, precision and throughput. All the results demonstrated that our developed fully automated platform should provide new opportunities to improve the accuracy, reproducibility, and throughput for proteome quantification.
语种	英语
WOS记录号	WOS:000404087600009
内容类型	期刊论文
源URL	[http://cas-ir.dicp.ac.cn/handle/321008/152111]
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
作者单位	1.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 2.Chinese Acad Sci, CAS Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
推荐引用方式 GB/T 7714	Yuan, Huiming,Zhang, Shen,Zhao, Baofeng,et al. Enzymatic Reactor with Trypsin Immobilized on Graphene Oxide Modified Polymer Microspheres To Achieve Automated Proteome Quantification[J]. ANALYTICAL CHEMISTRY,2017,89:6324-6329.
APA	Yuan, Huiming.,Zhang, Shen.,Zhao, Baofeng.,Weng, Yejing.,Zhu, Xudong.,...&Zhang, Yukui.(2017).Enzymatic Reactor with Trypsin Immobilized on Graphene Oxide Modified Polymer Microspheres To Achieve Automated Proteome Quantification.ANALYTICAL CHEMISTRY,89,6324-6329.
MLA	Yuan, Huiming,et al."Enzymatic Reactor with Trypsin Immobilized on Graphene Oxide Modified Polymer Microspheres To Achieve Automated Proteome Quantification".ANALYTICAL CHEMISTRY 89(2017):6324-6329.