Online Proteolysis and Glycopeptide Enrichment with Thermoresponsive Porous Polymer Membrane Reactors for Nanoflow Liquid Chromatography-Tandem Mass Spectrometry
文献类型：期刊论文
| 作者 | Yang, Joon Seon; Qiao, Juan; Kim, Jin Yong; Zhao, Liping; Qi, Li; Moon, Myeong Hee |
| 刊名 | ANALYTICAL CHEMISTRY
 |
| 出版日期 | MAR 6 2018 |
| 卷号 | 90期号:5页码:3124-3131 |
| 英文摘要 | In this Article, we have reported a fully automated online method to carry out proteolysis and glycopeptide enrichment in sequence for nanoflow liquid chromatography-tandem mass spectrometry (nLC-ESI-MS/MS) analysis. By implementing two serial thermoresponsive porous polymer membrane reactors (TPPMRs), in which the TPPM could be immobilized either with trypsin for proteolysis or with lectins for glycopeptide enrichment, the entire pretreatment procedure can be performed online in about an hour. The TPPM was fabricated by coating polystyrene-maleic anhydride-N-isopropylacrylamide (PS-MAn-PNIPAm), which was synthesized by reversible addition fragmentation chain transfer polymerization, on a Nylon sheet. Because of the thermoresponsive nature of PNIPAm, it formed micelle cavities and changed its morphology at elevated temperatures, resulting in enhanced interactions between the enzyme or lectins and the proteins/peptides flowing through the membrane. The performances of the TPPMs were evaluated by varying the temperature conditions and the amount of standard proteins, showing that both proteolysis and glycopeptide enrichment with online deglycosylation were highly efficient at 37 degrees C. The developed online serial TPPMRs-nLC-ESI-MS/MS method was applied to the human plasma sample (1.5 mu L) and a total of 262 N-glycopeptides could be identified from 155 glycoproteins. Thus, the present work demonstrates a fully automated high speed analytical protocol for online proteolysis and glycopeptide enrichment, which is extremely useful for analyzing small amounts of the proteome samples. |
| WOS记录号 | WOS:000427095700024 |
| 源URL | [http://ir.iccas.ac.cn/handle/121111/38723]  |
| 专题 | 化学研究所_其它 |
| 推荐引用方式 GB/T 7714 | Yang, Joon Seon,Qiao, Juan,Kim, Jin Yong,et al. Online Proteolysis and Glycopeptide Enrichment with Thermoresponsive Porous Polymer Membrane Reactors for Nanoflow Liquid Chromatography-Tandem Mass Spectrometry[J]. ANALYTICAL CHEMISTRY,MAR ,90(5):3124-3131. |
| APA | Yang, Joon Seon,Qiao, Juan,Kim, Jin Yong,Zhao, Liping,Qi, Li,&Moon, Myeong Hee.(MAR ).Online Proteolysis and Glycopeptide Enrichment with Thermoresponsive Porous Polymer Membrane Reactors for Nanoflow Liquid Chromatography-Tandem Mass Spectrometry.ANALYTICAL CHEMISTRY,90(5),3124-3131. |
| MLA | Yang, Joon Seon,et al."Online Proteolysis and Glycopeptide Enrichment with Thermoresponsive Porous Polymer Membrane Reactors for Nanoflow Liquid Chromatography-Tandem Mass Spectrometry".ANALYTICAL CHEMISTRY 90.5(MAR ):3124-3131. |
入库方式： OAI收割
来源：化学研究所
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