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Genetically biotinylated recombinant antibody in immunofiltration assay by light addressable potentiometric sensor for identification of Venezuelan equine encephalitis virus
Genetically biotinylated recombinant antibody in immunofiltration assay by light addressable potentiometric sensor for identification of Venezuelan equine encephalitis virus
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机译：光寻址电位传感器在免疫过滤法中生物遗传化重组抗体鉴定委内瑞拉马脑炎病毒
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摘要
A genetically biotinylated single chain fragment variable (scFv) antibody against Venezuelan equine encephalitis virus (VEE) being applied in a system consisting of an immunofiltration-enzyme assay (IFA) with a light addressable potentiometric sensor (LAPS) for the rapid identification of VEE is disclosed. The IFA entails formation of an immunocomplex sandwich consisting of VEE, biotinylated antibody, fluoresceinated antibody and streptavidin, capturing the sandwich by filtration on biotinylated membrane, and detecting the sandwich by anti-fluorescein urease conjugate. The concentration ratio of biotinylated to fluoresceinated antibodies is investigated and optimized. The IFA/LAPS assay sensitivity was approximately equal to that of a conventional enzyme-linked immunosorbant assay utilizing polystyrene plates and a chromogenic substrate, however, less time and effort were required for performance of the IFA/LAPS assay.
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机译：一种抗委内瑞拉马脑炎病毒（VEE）的遗传生物素化单链片段可变（scFv）抗体，已在由免疫过滤酶测定（IFA）和光寻址电位传感器（LAPS）组成的系统中使用，用于快速识别VEE。披露。 IFA需要形成由VEE，生物素化抗体，荧光素抗体和链霉亲和素组成的免疫复合物三明治，通过在生物素化膜上过滤来捕获三明治，并通过抗荧光素脲酶偶联物检测三明治。研究并优化了生物素化抗体与荧光素抗体的浓度比。 IFA / LAPS测定的灵敏度大约与使用聚苯乙烯板和发色底物的常规酶联免疫吸附测定的灵敏度相同，但是，执行IFA / LAPS测定所需的时间和精力更少。
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