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請使用永久網址來引用或連結此文件:
http://ir.lib.ncu.edu.tw/handle/987654321/28142
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題名: | Quantitative measurement of binding kinetics in sandwich assay using a fluorescence detection fiber-optic biosensor |
作者: | Lin,CH;Chen,HY;Yu,CJ;Lu,PL;Hsieh,CH;Hsieh,BY;Chang,YF;Chou,C |
貢獻者: | 光電科學研究所 |
關鍵詞: | POLY(METHYL METHACRYLATE);SURFACE;PROTEIN;SENSORS;IMMUNOSENSORS;ANTIGEN;PLASMA |
日期: | 2009 |
上傳時間: | 2010-06-29 19:42:24 (UTC+8) |
出版者: | 中央大學 |
摘要: | Fiber-optic biosensors have been Studied intensively because they are very useful and important tools for monitoring biomolecular interactions. Here we describe a fluorescence detection fiber-optic biosensor (FD-FOB) using a sandwich assay to detect antibody-antigen interaction. In addition, the quantitative measurement of binding kinetics, including the association and dissociation rate constants for immunoglobulin G (IgG)/anti-mouse IgG, is achieved, indicating 0.38 x 10(6), M-1 s(-1) for k(a) and 3.15 x 10(-3) s(-1) for k(d). These constants are calculated from the fluorescence signals detected oil fiber surface only where the excited evanescent wave can be generated. Thus, a confined fluorescence-detecting region is achieved to specifically determine the binding kinetics at the vicinity of the interface between sensing materials and uncladded fiber surface. With this FD-FOB, the mathematical deduction and experimental verification of the binding kinetics in a sandwich immunoassay provide a theoretical basis for measuring rate constants and equilibrium dissociation constants. A further measurement to study the interaction between human heart-type fatty acid-binding protein and its antibody gave the calculated kinetic constants k(a), k(d), and K-D as 8.48 x 10(5) M-1 s(-1), 1.7 x 10(-3) s(-1), and 2.0 nM, respectively. Our study is the first attempt to establish a theoretical basis for the florescence-sensitive immunoassay using a sandwich format. Moreover, we demonstrate that the FD-FOB as a high-throughput biosensor can provide an alternative to the chip-based biosensors to study real-time biomolecular interaction. (c) 2008 Elsevier Inc. All rights reserved. |
關聯: | ANALYTICAL BIOCHEMISTRY |
顯示於類別: | [光電科學研究所] 期刊論文
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