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Immunoassay Based on Electroge Nerated Chemiluminescence Amplified by Magnetic Nanobeads and Electrochemiluminescence Resonance Energy Transfer
Author: LiMingYue
Tutor: ZhangXiaoLi
School: Shandong University
Course: Analytical Chemistry
Keywords: Electrochemiluminescence Immunoassays Submicron magnetic ball Electrochemiluminescence resonance energy transfer DNA analysis Quantum dots Luminol
CLC: O657.1
Type: Master's thesis
Year: 2010
Downloads: 170
Quote: 1
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Abstract
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The first chapter, as ECL of Ru ( bpy ) 2 ( dcbpy ) NHS markers will be amplified electrochemical luminescence immunoassay analysis and magnetic ball technology, the development of an ultra- sensitive detection of antigen and antibody electrochemical luminescence detection method . The method submicron magnetic ball surface modification of a large number of light - emitting molecules play a role in signal amplification . Experiment we use the surface of the antibody amino coverslip substrate reaction of silylated epoxy group ( s ) of the antibody fixed in the surface of the substrate , and then by the immune response , biotin and streptavidin- biotin -specific reactions eventually formed streptavidin and biotinylated magnetic ball - biotinylated antibody - antigen - antibody double -antibody sandwich complexes, and dissociation . ( Dcbpy ) NHS ester group on Ru ( bpy ) with magnetic ball on streptavidin- biotin binding reaction between the ECL markers mark to the surface of the magnetic ball , the modified experiment ECL markers magnetic ball a magnet fixed to the surface of the Au electrode . ECL detection in the presence of tripropylamine , 1.35 V maximum the electrochemical luminescence intensity ( IM , ECL ) . With IM , ECL , we can achieve the detection of the antigen . The use of streptavidin- biotin - modified magnetic balls ( SA - SMBs ) can increase the quantity of Ru ( bpy ) 32 corresponding to a single target molecule molecules , obtained through the signal amplification effect , greatly improving the electrochemical oxidation of Ru ( bpy ) 32 molecules second ECL wave , thereby improving the detection sensitivity of the method . Using this method, we realize the detection of CEA , the linear range of 1.0 x 10-14 mol / L to 3.0 × 10 - 13 mol / L . In the second chapter , we introduce electrochemiluminescence as a donor light source of a new resonance energy transfer system - electrochemiluminescence resonance energy transfer ( ECRET ) . Electrochemiluminescence reagents , issued by the electrochemical reaction of a center wavelength of 460 nm blue light serves as an energy donor in the presence of hydrogen peroxide under conditions isoluminol ; addition, select a maximum emission peak at 655 nm of the red fluorescent quantum dots as energy acceptor . When applying a potential to the electrode , electrochemical light emitting donor energy transfer to a receptor of quantum dots , the efficient electrochemical luminescence resonance energy transfer. The ECRET techniques can be used to study the interactions between biological molecules , DNA analysis and immunological analysis of nucleic acids .
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CLC: > Mathematical sciences and chemical > Chemistry > Analytical Chemistry > Instrument analysis ( physics and physical chemistry ) > Electrochemical analysis
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