PMID: 9433463Jan 20, 1998Paper

A comparative study by the enzyme-linked immunofiltration assay of solid phases used in the development of flow immunosensors

Journal of Immunological Methods
S MoraisR Puchades

Abstract

The application of an inert membrane-based, enzyme-linked immunofiltration assay (ELIFA) to the characterization of immunosorbents suitable for flow immunosensor development is described. For direct assays, eight monoclonal antibodies (MAb) raised against the insecticide carbaryl were immobilized on three sorbents, namely, controlled pore glass (CPG), hydrazide derivatized agarose beads and a hydrophilic polymer with immobilized Protein A/G. The interaction between immobilized antibodies and antigen was directly detected using a carbaryl hapten conjugated to horseradish peroxidase. Immunosorbent characterization was based on both sensitivity and re-usability. Optimal immunosorbent regeneration was achieved using 0.1 M glycine/HCl, pH 2.0 as the desorbent solution. The best covalent immunosorbent was obtained by immobilizing LIB-CNA36 MAb on hydrazide derivatized agarose beads. The best immunosorbent obtained by reversible immobilization was LIB-CNH45 MAb on Protein A/G. Using this support the eventual irreversible denaturation of covalently immobilized MAbs was overcome. For indirect assays, N-hydroxisuccinimide derivatized agarose beads and glutaraldehyde-activated CPG were used as sorbents for hapten immobilization via the am...Continue Reading

References

Apr 1, 1983·Analytical Chemistry·J Sherma, G Zweig

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Citations

Mar 3, 2005·Biosensors & Bioelectronics·Pierre N FlorianoJohn T McDevitt
Oct 31, 2006·Analytical and Bioanalytical Chemistry·M A González-MartínezA Maquieira
Oct 20, 2009·The Analyst·Cristina García-AljaroEva Baldrich
Mar 6, 1999·Journal of Chromatography. B, Biomedical Sciences and Applications·J Turková
Apr 5, 2001·Biosensors & Bioelectronics·J PenalvaB D Hammock
Sep 18, 1999·Analytical Chemistry·J PenalvaA Maquieira

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