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题名: Determination of binding constants between one protein and multiple carbohydrates by affinity chromatography on a microchip
作者: Liu, Xiaojun1;  Wang, Hui2;  Liang, Aiye3;  Li, Yuling1;  Gai, Hongwei1;  Lin, Bingcheng4
关键词: Binding constants ;  Carbohydrate-protein interaction ;  Drug screening ;  Affinity chromatography ;  Microchip capillary electrophoresis
刊名: JOURNAL OF CHROMATOGRAPHY A
发表日期: 2012-12-28
DOI: 10.1016/j.chroma.2012.10.042
卷: 1270, 页:340-343
收录类别: SCI
文章类型: Article
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: Development of rapid, reliable and high throughput methods for evaluating the interactions between different carbohydrates and a same protein is critical to carbohydrate drug development. In this study, we develop a novel strategy based on an affinity chromatography for quickly determining the binding constants of different carbohydrates to a same protein. The core of our method is the inversely proportional relationship between the binding constant and a new termed parameter, critical elution concentration (CMC). CMC is defined as the lowest concentration of displacing reagent, a series of carbohydrates herein, at which the protein specifically bond to the affinity column can be eluted off as an intact peak by the carbohydrate solution in a certain time. The interactions between a series of sulfate polysaccharides and granulocyte colony-stimulating factor (G-CSF) are selected as model. Through a 200 mu m long heparin affinity column microfabricated inside a channel of 50 mu m width and 20 mu m height, the binding constant of each G-CSF-polysaccharide binding pair can be obtained within 1 h, around one sixth of time needed by traditional capillary electrophoresis based method. (C) 2012 Elsevier B.V. All rights reserved.
关键词[WOS]: COLONY-STIMULATING FACTOR ;  SURFACE-PLASMON RESONANCE ;  CAPILLARY-ELECTROPHORESIS ;  MICROFLUIDIC CHIP ;  HEPARIN ;  SEPARATION ;  OLIGOSACCHARIDES ;  GLYCANS ;  SULFATE ;  DESIGN
语种: 英语
WOS记录号: WOS:000312572900041
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/142903
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.Jiangsu Normal Univ, Sch Chem & Engn, Xuzhou 221116, Peoples R China
2.China Pharmaceut Univ Pharmaceut Co Ltd, Nanjing 210009, Jiangsu, Peoples R China
3.Charleston So Univ, Dept Phys Sci, N Charleston, SC 29406 USA
4.Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China

Recommended Citation:
Liu, Xiaojun,Wang, Hui,Liang, Aiye,et al. Determination of binding constants between one protein and multiple carbohydrates by affinity chromatography on a microchip[J]. JOURNAL OF CHROMATOGRAPHY A,2012,1270:340-343.
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