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题名: On-line combination of capillay isoelectric focusing and capillary non-gel sieving electrophoresis using a hollow-fiber membrane interface: a novel two-dimensional separation system for proteins
作者: Liu, HC;  Yang, C;  Yang, Q;  Zhang, WB;  Zhang, YK
关键词: capillary isoelectric focusing ;  capillary non-gel sieving electrophoresis ;  hollow-fiber membrane interface ;  proteins
刊名: JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
发表日期: 2005-03-05
DOI: 10.1016/j.jchromb.2004.04.038
卷: 817, 期:1, 页:119-126
收录类别: SCI
文章类型: Article
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
英文摘要: A novel two-dimensional (2D) separation system for proteins was reported. In the system, a piece of dialysis hollow-fiber membrane was employed as the interface for on-line combination of capillary isoelectric focusing (CIEF) and capillary non-gel sieving electrophoresis (CNGSE). The system is similar equivalent to two-dimensional polyacrylamide gel electrophoresis (2D PAGE), by transferring the principal of 2D PAGE separation to the capillary format. Proteins were focused and separated in first dimension CIEF based on their differences in isoelectric points (pIs). Focused protein zones was transferred to the dialysis hollow-fiber interface, where proteins hydrophobically complexed with sodium dodecyl sulfate (SDS). The negatively charged proteins were electromigrated and further resolved by their differences in size in the second dimension CNGSE, in which dextran solution, a replaceable sieving matrix instead of cross-linked polyacrylamide gel was employed for size-dependent separation of proteins. The combination of the two techniques was attributed to high efficiency of the dialysis membrane interface. The feasibility and the orthogonality of the combined CIEF-CNGSE separation technique, an important factor for maximizing peak capacity or resolution elements, were demonstrated by examining each technique independently for the separation of hemoglobin and protein mixtures excreting from lung cancer cells of rat. The 2D separation strategy was found to greatly increase the resolving power and overall peak capacity over those obtained for either dimension alone. (C) 2004 Elsevier B.V. All rights reserved.
关键词[WOS]: HIGH-PERFORMANCE ELECTROPHORESIS ;  PHASE LIQUID-CHROMATOGRAPHY ;  ZONE-ELECTROPHORESIS ;  MASS-SPECTROMETRY ;  POLYACRYLAMIDE-GEL ;  PEPTIDES ;  PROTEOMICS ;  DEVICE
语种: 英语
WOS记录号: WOS:000226919100013
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/139933
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, Natl Chromatograph R&A Ctr, Dalian Inst Chem Phys, Dalian 116011, Peoples R China

Recommended Citation:
Liu, HC,Yang, C,Yang, Q,et al. On-line combination of capillay isoelectric focusing and capillary non-gel sieving electrophoresis using a hollow-fiber membrane interface: a novel two-dimensional separation system for proteins[J]. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,2005,817(1):119-126.
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