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| Measuring Binding Kinetics Of Surface-Bound Molecules Using The Surface Plasmon Resonance Technique | |
Li BX(李宝霞) ; Chen J(陈娟); Long M(龙勉); Long, M (reprint author), Chinese Acad Sci, Inst Mech, Natl Micrograv Lab, Beijing 100190, Peoples R China.
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| Source Publication | Analytical Biochemistry
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| 2008 | |
| Pages | 195-201 |
| ISSN | 0003-2697 |
| Abstract | Surface plasmon resonance (SPR) technology and the Biacore biosensor have been widely used to measure the kinetics of biomolecular interactions in the fluid phase. In the past decade, the assay was further extended to measure reaction kinetics when two counterpart molecules are anchored on apposed surfaces. However, the cell binding kinetics has not been well quantified. Here we report development of a cellular kinetic model, combined with experimental procedures for cell binding kinetic measurements, to predict kinetic rates per cell. Human red blood cells coated with bovine serum albumin and anti-BSA monoclonal antibodies (mAbs) immobilized on the chip were used to conduct the measurements. Sensor-grams for BSA-coated RBC binding onto and debinding from the anti-BSA mAb-immobilized chip were obtained using a commercial Biacore 3000 biosensor, and analyzed with the cellular kinetic model developed. Not only did the model fit the data well, but it also predicted cellular on and off-rates as well as binding affinities from curve fitting. The dependence of flow duration, flow rate, and site density of BSA on binding kinetics was tested systematically, which further validated the feasibility and reliability of the new approach. Crown copyright (c) 2008 Published by Elsevier Inc. All rights reserved. |
| Keyword | Cellular Off-rate Cellular On-rate Response Unit Kinetic Model Optical Biosensor Literature Selectin Antibody Biacore Antigen Constants Transport Protein Cells Shear |
| DOI | 10.1016/j.ab.2008.03.030 |
| Indexed By | SCI |
| Language | 英语 |
| WOS ID | WOS:000255918000013 |
| WOS Keyword | OPTICAL BIOSENSOR LITERATURE ; SELECTIN ; ANTIBODY ; BIACORE ; ANTIGEN ; CONSTANTS ; TRANSPORT ; PROTEIN ; CELLS ; SHEAR |
| WOS Research Area | Biochemistry & Molecular Biology ; Chemistry |
| WOS Subject | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Analytical |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://dspace.imech.ac.cn/handle/311007/26038 |
| Collection | 力学所知识产出(1956-2008) |
| Corresponding Author | Long, M (reprint author), Chinese Acad Sci, Inst Mech, Natl Micrograv Lab, Beijing 100190, Peoples R China. |
| Recommended Citation GB/T 7714 | Li BX,Chen J,Long M,et al. Measuring Binding Kinetics Of Surface-Bound Molecules Using The Surface Plasmon Resonance Technique[J]. Analytical Biochemistry,2008:195-201. |
| APA | 李宝霞,陈娟,龙勉,&Long, M .(2008).Measuring Binding Kinetics Of Surface-Bound Molecules Using The Surface Plasmon Resonance Technique.Analytical Biochemistry,195-201. |
| MLA | 李宝霞,et al."Measuring Binding Kinetics Of Surface-Bound Molecules Using The Surface Plasmon Resonance Technique".Analytical Biochemistry (2008):195-201. |
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