DICP OpenIR
Bridged Hybrid Monolithic Column Coupled to High-Resolution Mass Spectrometry for Top-Down Proteomics
Liang, Yu1,2; Jin, Yutong3; Wu, Zhijie3; Tucholski, Trisha3; Brown, Kyle A.3; Zhang, Lihua2; Zhang, Yukui2; Ge, Ying1,3,4
Corresponding AuthorZhang, Lihua(lihuazhang@dicp.ac.cn) ; Ge, Ying(ying.ge@wisc.edu)
Source PublicationANALYTICAL CHEMISTRY
2019-02-05
ISSN0003-2700
DOI10.1021/acs.analchem.8b05817
Volume91Issue:3Pages:1743-1747
Funding Projecthigh-end instrument Grant[S10OD018475] ; NIH R01 Grants[GM117058] ; NIH R01 Grants[GM125085] ; NIH R01 Grants[HL109810] ; NIH R01 Grants[HL096971] ; China Scholarship Council ; National Natural Science Foundation of China[21575139]
Funding Organizationhigh-end instrument Grant ; high-end instrument Grant ; NIH R01 Grants ; NIH R01 Grants ; China Scholarship Council ; China Scholarship Council ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; high-end instrument Grant ; high-end instrument Grant ; NIH R01 Grants ; NIH R01 Grants ; China Scholarship Council ; China Scholarship Council ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; high-end instrument Grant ; high-end instrument Grant ; NIH R01 Grants ; NIH R01 Grants ; China Scholarship Council ; China Scholarship Council ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; high-end instrument Grant ; high-end instrument Grant ; NIH R01 Grants ; NIH R01 Grants ; China Scholarship Council ; China Scholarship Council ; National Natural Science Foundation of China ; National Natural Science Foundation of China
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordHYDROPHOBIC INTERACTION CHROMATOGRAPHY ; SIZE-EXCLUSION CHROMATOGRAPHY ; EFFECTIVE PROTEIN SEPARATION ; LIQUID-CHROMATOGRAPHY ; PHASE CHROMATOGRAPHY ; INTACT PROTEINS ; Z-DISC ; POLYMERIZATION ; TECHNOLOGY
AbstractTop-down mass spectrometry (MS)-based proteomics has become a powerful tool for comprehensive characterization of intact proteins. However, because of the high complexity of the proteome, highly effective separation of intact proteins from complex mixtures prior to MS analysis remains challenging. Monolithic columns have shown great promise for intact protein separation due to their high permeability, low backpressure, and fast mass transfer. Herein, for the first time, we developed bridged hybrid bis(triethoxysilyl)ethylene (BTSEY) monolith with C8 functional groups (C8@BTSEY) for highly effective protein separation and coupled it to high-resolution MS for identification of intact proteins from complex protein mixtures. We have optimized mobile phase conditions of our monolith -based reverse-phase chromatography (RPC) for online liquid chromatography (LC)-MS analysis and evaluated separation reproducibility of the C8pBTSEY column. We further assessed the chromatographic performance of this column by separating a complex protein mixture extracted from swine heart tissue. Using our monolithic column (i.d. 100 mu m X 35 cm), we separated over 300 proteoforms (up to 104 kDa) from 360 ng of protein mixture in an 80 min one-dimensional (1D) LC run. The highly effective separation and recovery of intact proteins from this monolithic column allowed unambiguous identification of similar to 100 proteoforms including a large protein, alpha actinin2 (103.77 kDa), by online ID LC-MS/MS-analysis for the first time. As demonstrated, this C8pBTSEY column is reproducible and effective in separation of intact proteins, which shows high promise for top-down proteomics.
Language英语
Funding Organizationhigh-end instrument Grant ; high-end instrument Grant ; NIH R01 Grants ; NIH R01 Grants ; China Scholarship Council ; China Scholarship Council ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; high-end instrument Grant ; high-end instrument Grant ; NIH R01 Grants ; NIH R01 Grants ; China Scholarship Council ; China Scholarship Council ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; high-end instrument Grant ; high-end instrument Grant ; NIH R01 Grants ; NIH R01 Grants ; China Scholarship Council ; China Scholarship Council ; National Natural Science Foundation of China ; National Natural Science Foundation of China ; high-end instrument Grant ; high-end instrument Grant ; NIH R01 Grants ; NIH R01 Grants ; China Scholarship Council ; China Scholarship Council ; National Natural Science Foundation of China ; National Natural Science Foundation of China
WOS IDWOS:000458220300015
PublisherAMER CHEMICAL SOC
Citation statistics
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/166109
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhang, Lihua; Ge, Ying
Affiliation1.Univ Wisconsin, Dept Cell & Regenerat Biol, Madison, WI 53705 USA
2.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Liaoning, Peoples R China
3.Univ Wisconsin, Dept Chem, Madison, WI 53706 USA
4.Univ Wisconsin, Sch Med & Publ Hlth, Human Prote Program, Madison, WI 53705 USA
Recommended Citation
GB/T 7714
Liang, Yu,Jin, Yutong,Wu, Zhijie,et al. Bridged Hybrid Monolithic Column Coupled to High-Resolution Mass Spectrometry for Top-Down Proteomics[J]. ANALYTICAL CHEMISTRY,2019,91(3):1743-1747.
APA Liang, Yu.,Jin, Yutong.,Wu, Zhijie.,Tucholski, Trisha.,Brown, Kyle A..,...&Ge, Ying.(2019).Bridged Hybrid Monolithic Column Coupled to High-Resolution Mass Spectrometry for Top-Down Proteomics.ANALYTICAL CHEMISTRY,91(3),1743-1747.
MLA Liang, Yu,et al."Bridged Hybrid Monolithic Column Coupled to High-Resolution Mass Spectrometry for Top-Down Proteomics".ANALYTICAL CHEMISTRY 91.3(2019):1743-1747.
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