文献详情
Development of dual-color total internal reflection fluorescence biosensor for simultaneous quantitation of two small molecules and their affinity constants with antibodies
文献类型：期刊
期刊名称：Biosensors & bioelectronics影响因子和分区
年：2019
卷：126
页码：824-830
ISSN：1873-4235
关键词：Affinity constant,Biosensor,Dual-color fluorescence,Small molecule,Total internal reflection
摘要：A novel dual-color total internal reflection fluorescence biosensor (DTB) was successfully developed for the simultaneous detection of two small molecules based on a simple optical structure and the time resolved effect of fiber optic switch. The DTB employed a single-multi mode fiber optic coupler instead of a sophisticated confocal optical system for the transmission of two excitation lights and dual-color fluorescence, and a photodiode detector instead of photomultiplier for the simultaneous ...More
A novel dual-color total internal reflection fluorescence biosensor (DTB) was successfully developed for the simultaneous detection of two small molecules based on a simple optical structure and the time resolved effect of fiber optic switch. The DTB employed a single-multi mode fiber optic coupler instead of a sophisticated confocal optical system for the transmission of two excitation lights and dual-color fluorescence, and a photodiode detector instead of photomultiplier for the simultaneous detection of dual-color fluorescence. The compact optical design of DTB improved its optical transmission efficiency and detection sensitivity because of no requirement of numerous optical separation elements and rigorous optical alignment. The DTB was applied for the simultaneous detection of 2,4-Bisphenol-A (BPA) and 2,4-Dichlorophenoxyacetic acid (2,4-D) using one bifunctional fiber optic bio-probe modified by two hapten-protein conjugates. When the mixture of Cy5.5 labeled anti-2,4-D antibody and Pacific Blue dye labeled anti-BPA antibody was introduced over the surface of the bio-probe, they bound with their respective hapten-protein conjugate immobilized onto the bio-probe. Based on the time-resolved effect of fiber optic switch, two fluorescence dyes were alternatively excited by 635?nm and 405?nm laser lights and simultaneously detected by one photodiode detector. Taking indirect competitive immunoassay principle, BPA and 2,4-D were simultaneously detected using the DTB with high sensitivity, accuracy, and rapidity. The quantitation of affinity constants between small molecules and their antibodies was also achieved based on the proposed theory. The DTB provides a flexible and powerful platform for simultaneously sensitive quantitation of multiple targets and the affinity constants of biomolecular interactions.Copyright ? 2018 Elsevier B.V. All rights reserved. ...Hide

DOI：10.1016/j.bios.2018.12.010
百度学术：Development of dual-color total internal reflection fluorescence biosensor for simultaneous quantitation of two small molecules and their affinity constants with antibodies
语言：外文
人气指数：3
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