GENETICALLY BIOTINYLATED RECOMBINANT ANTIBODY IN IMMUNOFILTRATION ASSAY BY LIGHT ADDRESSABLE POTENTIOMETRIC SENSOR FOR IDENTIFICATION OF VENEZUELAN EQUINE ENCEPHALITIS VIRUS

摘要

A genetically biotinylated single chain fragment variable (scFv) antibodyagainstVenezuelan equine encephalitis virus (VEE) being applied in a systemconsisting of animmunofiltration-enzyme assay (IFA) with a light addressable potentiometricsensor(LAPS) for the rapid identification of VEE is disclosed. The IFA entailsformation of animmunocomplex sandwich consisting of VEE, biotinylated antibody,fluoresceinatedantibody and streptavidin, capturing the sandwich by filtration onbiotinylated membrane,and detecting the sandwich by anti-fluorescein urease conjugate. Theconcentration ratioof biotinylated to fluoresceinated antibodies is investigated and optimized.TheIFA/LAPS assay sensitivity was approximately equal to that of a conventionalenzyme-linked immunosorbant assay utilizing polystyrene plates and a chromogenicsubstrate,however, less time and effort were required for performance of the IFA/LAPSassay.