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学科主题: 物理化学
题名: High sensitive protein detection by hollow fiber membrane interface based protein enrichment and in situ fluorescence derivatization
作者: Qiao, Xiaoqiang1, 2;  Sun, Liangliang1, 2;  Wang, Li3;  Liang, Yu1, 2;  Zhang, Lihua1;  Shan, Yichu1;  Peng, Xiaojun3;  Liang, Zhen1;  Zhang, Yukui1
通讯作者: 张丽华
关键词: In situ fluorescence derivatization ;  Hollow fiber membrane interface ;  Preconcentration ;  Low abundance proteins
刊名: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
发表日期: 2011-05-15
DOI: 10.1016/j.jchromb.2010.11.002
卷: 879, 期:17-18, 页:1439-1443
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1810
产权排名: 1,1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
摘要: High sensitive protein detection by hollow fiber membrane interface based protein enrichment and in situ fluorescence derivatization
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: A novel device, composed of a syringe pump for sample loading, a hydrophilic hollow fiber membrane interface for protein concentration and small molecules removal, and a centrifugation tube for buffer exchange, was designed for protein preconcentration and in situ fluorescence derivatization. With the outlet of the interface blocked, denatured proteins were continually introduced. Restricted by the membrane with the molecular weight cutoff (MWCO) of 3000 Da, proteins were concentrated within the membrane. However, denaturant and other small molecules, which might affect the further fluorescence derivatization, were driven out of the membrane. Then, the membrane with proteins restricted inside was directly put into the fluorescence derivatization buffer. Here, the water-soluble sulfo-3H-indocyanine dye, the active N-hydroxysuccinimide of 3H-indolium,1-(5-carboxypentyl)-2[3-[1-(5-carboxypentyl)-1,3-dihydro-3,3-dimethyl-5-sulfo-2H-indol-2-ylidene]-1-propenyl]-3,3-dimethy1-5-sulfo-,monopotassium salt (sw-cy3-NHS). synthesized in our lab. was used for protein labeling. By such a method, the detection sensitivity of bovine serum albumin (BSA) was improved by nearly 200 folds, compared to that obtained by direct in-solution derivatization. Through the derivatization of a fraction of E. colt protein separated by reversed phase HPLC, proteins with low concentration were efficiently labeled, which indicated the potential merit of the developed method for the high sensitive detection of low abundance proteins. (C) 2010 Elsevier B.V. All rights reserved.
关键词[WOS]: SODIUM DODECYL-SULFATE ;  PERFORMANCE LIQUID-CHROMATOGRAPHY ;  CAPILLARY GEL-ELECTROPHORESIS ;  TANDEM MASS-SPECTROMETRY ;  NATIVE PROTEINS ;  AMINO-ACIDS ;  PRECONCENTRATION ;  SEPARATION ;  IDENTIFICATION ;  PEPTIDES
语种: 英语
WOS记录号: WOS:000291131900027
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/115307
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
3.Dalian Univ Technol, State Key Lab Fine Chem, Dalian 116012, Peoples R China

Recommended Citation:
Qiao, Xiaoqiang,Sun, Liangliang,Wang, Li,et al. High sensitive protein detection by hollow fiber membrane interface based protein enrichment and in situ fluorescence derivatization[J]. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,2011,879(17-18):1439-1443.
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