| Quantitative measurement of binding kinetics in sandwich assay using a fluorescence detection fiber-optic biosensor. | |
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MedLine Citation:
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PMID: 19041630 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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 on 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. |
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Authors:
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Chao-Hsiung Lin; Hsaing-Yin Chen; Chih-Jen Yu; Pen-Li Lu; Chu-Hsin Hsieh; Bao-Yu Hsieh; Ying-Feng Chang; Chien Chou |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2008-11-18 |
Journal Detail:
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Title: Analytical biochemistry Volume: 385 ISSN: 1096-0309 ISO Abbreviation: Anal. Biochem. Publication Date: 2009 Feb |
Date Detail:
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Created Date: 2009-01-19 Completed Date: 2009-03-10 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0370535 Medline TA: Anal Biochem Country: United States |
Other Details:
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Languages: eng Pagination: 224-8 Citation Subset: IM |
Affiliation:
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Department of Life Sciences and Institute of Genome Sciences, National Yang Ming University, Taipei, Taiwan. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Antibodies Antigen-Antibody Reactions* Biosensing Techniques / methods* Fatty Acid-Binding Proteins / immunology Fluorescence Heart Humans Immunoassay / methods* Kinetics Myocardium / chemistry Optical Fibers |
| Chemical | |
Reg. No./Substance:
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0/Antibodies; 0/Fatty Acid-Binding Proteins |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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