Bridged hybrid monolithic column coupled to high-resolution mass spectrometry for top-down proteomics
文献类型：期刊论文
| 作者 | Liang, Yu1,2; Jin, Yutong3; Wu, Zhijie3; Tucholski, Trisha3; Brown, Kyle A.3; Zhang, Lihua2; Zhang, Yukui2; Ge, Ying1,3,4 |
| 刊名 | Analytical chemistry
 |
| 出版日期 | 2019-02-05 |
| 卷号 | 91期号:3页码:1743-1747 |
| ISSN号 | 0003-2700 |
| DOI | 10.1021/acs.analchem.8b05817 |
| 通讯作者 | Zhang, lihua(lihuazhang@dicp.ac.cn) ; Ge, ying(ying.ge@wisc.edu) |
| 英文摘要 | Top-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. |
| WOS关键词 | HYDROPHOBIC INTERACTION CHROMATOGRAPHY ; SIZE-EXCLUSION CHROMATOGRAPHY ; EFFECTIVE PROTEIN SEPARATION ; LIQUID-CHROMATOGRAPHY ; PHASE CHROMATOGRAPHY ; INTACT PROTEINS ; Z-DISC ; POLYMERIZATION ; TECHNOLOGY |
| WOS研究方向 | Chemistry |
| WOS类目 | Chemistry, Analytical |
| 语种 | 英语 |
| 出版者 | AMER CHEMICAL SOC |
| WOS记录号 | WOS:000458220300015 |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2372821 |
| 专题 | 大连化学物理研究所 |
| 通讯作者 | Zhang, Lihua; Ge, Ying |
| 作者单位 | 1.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 |
| 推荐引用方式 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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来源：大连化学物理研究所
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