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Enzymatic Reactor with Trypsin Immobilized on Graphene Oxide Modified Polymer Microspheres To Achieve Automated Proteome Quantification
Yuan, Huiming1; Zhang, Shen1,2; Zhao, Baofeng1; Weng, Yejing1,2; Zhu, Xudong1,2; Li, Senwu1,2; Zhang, Lihua1; Zhang, Yukui1
刊名ANALYTICAL CHEMISTRY
2017-06-20
DOI10.1021/acs.analchem.7b00682
89页:6324-6329
收录类别SCI
文章类型Article
WOS标题词Science & Technology ; Physical Sciences
类目[WOS]Chemistry, Analytical
研究领域[WOS]Chemistry
关键词[WOS]HIGHLY EFFICIENT ; QUANTITATIVE PROTEOMICS ; MASS-SPECTROMETRY ; DIGESTION ; METHODOLOGY ; COMBINATION ; STRATEGIES ; PROTEINS ; MIXTURES ; CANCER
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
引用统计
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/152111
专题中国科学院大连化学物理研究所
作者单位1.Chinese Acad Sci, CAS Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
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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.
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