DICP OpenIR
学科主题物理化学
High sensitive protein detection by hollow fiber membrane interface based protein enrichment and in situ fluorescence derivatization
Qiao, Xiaoqiang1,2; Sun, Liangliang1,2; Wang, Li3; Liang, Yu1,2; Zhang, Lihua1; Shan, Yichu1; Peng, Xiaojun3; Liang, Zhen1; Zhang, Yukui1; Zhang LH(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
2011-05-15
ISSN待补充
DOI10.1016/j.jchromb.2010.11.002
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
研究领域[WOS]Biochemistry & Molecular Biology ; Chemistry
关键词[WOS]SODIUM DODECYL-SULFATE ; PERFORMANCE LIQUID-CHROMATOGRAPHY ; CAPILLARY GEL-ELECTROPHORESIS ; TANDEM MASS-SPECTROMETRY ; NATIVE PROTEINS ; AMINO-ACIDS ; PRECONCENTRATION ; SEPARATION ; IDENTIFICATION ; PEPTIDES
英文摘要High sensitive protein detection by hollow fiber membrane interface based protein enrichment and in situ fluorescence derivatization; 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.
语种英语
WOS记录号WOS:000291131900027
引用统计
被引频次:5[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/115307
专题中国科学院大连化学物理研究所
通讯作者Zhang LH(张丽华)
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
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GB/T 7714
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.
APA Qiao, Xiaoqiang.,Sun, Liangliang.,Wang, Li.,Liang, Yu.,Zhang, Lihua.,...&张丽华.(2011).High sensitive protein detection by hollow fiber membrane interface based protein enrichment and in situ fluorescence derivatization.JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,879(17-18),1439-1443.
MLA Qiao, Xiaoqiang,et al."High sensitive protein detection by hollow fiber membrane interface based protein enrichment and in situ fluorescence derivatization".JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 879.17-18(2011):1439-1443.
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