Development of immunofiltration assay by light addressable potentiometric sensor with genetically biotinylated recombinant antibody for rapid identification of Venezuelan equine encephalitis virus

Journal of Immunological Methods
Wei-Gang HuR Elaine Fulton

Abstract

A genetically biotinylated single chain fragment variable antibody (scFv) against Venezuelan equine encephalitis virus (VEE) was applied in a system consisting of an immunofiltration enzyme assay (IFA) with a light addressable potentiometric sensor (LAPS) for the rapid identification of VEE. The IFA involved formation of an immunocomplex sandwich consisting of VEE, biotinylated antibody, fluoresceinated antibody and streptavidin, capture of the sandwich by filtration on biotinylated membrane, and labeling of the sandwich by anti-fluorescein urease conjugate. The concentration ratio of biotinylated to fluoresceinated antibodies was investigated and optimized. By the IFA/LAPS assay, the limit of detection (LOD) of VEE was approximately 30 ng/ml, similar to that achieved when chemically biotinylated monoclonal antibody (mAb) was applied. Total assay variance of the IFA/LAPS assay for both intra- and inter-assay precision was less than 20%. Assay accuracy was measured by comparing VEE concentrations estimated by IFA/LAPS standard curve to those obtained by conventional protein assay. VEE concentrations were found to differ by no more than 10%. The IFA/LAPS assay sensitivity was approximately equal to that of a conventional enzyme-l...Continue Reading

References

Aug 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·J S HustonR Crea
Nov 5, 1993·Journal of Immunological Methods·W E LeeJ P Wong
Aug 6, 1997·JAMA : the Journal of the American Medical Association·G W ChristopherE M Eitzen
Nov 26, 1998·Journal of Biotechnology·D LuoR Madiyalakan
Aug 17, 2000·Biosensors & Bioelectronics·P A EmanuelA Dion-Schultz

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Citations

Dec 14, 2011·Analytical and Bioanalytical Chemistry·Xiangqun ZengRay Mernaugh
Apr 28, 2009·Journal of Microbiological Methods·Jason E DoverSang-Jin Suh
May 12, 2009·Seminars in Cell & Developmental Biology·Paul J ConroyRichard J O'Kennedy
Sep 28, 2006·The Analyst·Bobby PejcicGordon Parkinson
Jun 8, 2017·Proceedings. Mathematical, Physical, and Engineering Sciences·Fan WuSteffi Krause
Feb 6, 2020·Reproductive Sciences·Dorien OThomas D'Hooghe
Apr 4, 2013·Bioorganicheskaia khimiia·S I RomaniukS V Komisarenko
Apr 26, 2021·Biosensors & Bioelectronics·Hassan Karimi-MalehAmani Al-Othman

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