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Determination of binding constants between one protein and multiple carbohydrates by affinity chromatography on a microchip
Liu, Xiaojun1; Wang, Hui2; Liang, Aiye3; Li, Yuling1; Gai, Hongwei1; Lin, Bingcheng4
KeywordBinding Constants Carbohydrate-protein Interaction Drug Screening Affinity Chromatography Microchip Capillary Electrophoresis
Source PublicationJOURNAL OF CHROMATOGRAPHY A
2012-12-28
DOI10.1016/j.chroma.2012.10.042
Volume1270Pages:340-343
Indexed BySCI
SubtypeArticle
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS KeywordCOLONY-STIMULATING FACTOR ; SURFACE-PLASMON RESONANCE ; CAPILLARY-ELECTROPHORESIS ; MICROFLUIDIC CHIP ; HEPARIN ; SEPARATION ; OLIGOSACCHARIDES ; GLYCANS ; SULFATE ; DESIGN
AbstractDevelopment 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.
Language英语
WOS IDWOS:000312572900041
Citation statistics
Cited Times:6[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/142903
Collection中国科学院大连化学物理研究所
Affiliation1.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
GB/T 7714
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.
APA Liu, Xiaojun,Wang, Hui,Liang, Aiye,Li, Yuling,Gai, Hongwei,&Lin, Bingcheng.(2012).Determination of binding constants between one protein and multiple carbohydrates by affinity chromatography on a microchip.JOURNAL OF CHROMATOGRAPHY A,1270,340-343.
MLA Liu, Xiaojun,et al."Determination of binding constants between one protein and multiple carbohydrates by affinity chromatography on a microchip".JOURNAL OF CHROMATOGRAPHY A 1270(2012):340-343.
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