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Title

Fast electrochemical detection of anti-HIV antibodies: Coupling allosteric enzymes and disk microelectrode arrays

AuthorsLaczka, Olivier; Ferraz, Rosa M.; Ferrer-Miralles, Neus; Villaverde Corrales, Antonio Pedro; Muñoz Pascual, Francisco Javier; Campo García, Francisco Javier del
KeywordsAllosteric enzymes
Biosensors
Microelectrode arrays
Electrochemistry
Amperometry
Coulometry
HIV
AIDS
Issue Date17-Mar-2009
CitationAnalytica Chimica Acta 641(1-2): 1-6 (2009)
AbstractHere a novel electrochemical method for the rapid detection of anti-HIV antibodies in serum is presented. The novelty lies in the combination of allosteric enzymes and coulometry to yield a fast, simple and reliable HIV diagnostic method. We have used a previously developed β-galactosidase enzyme that is efficiently activated by anti-HIV antibodies directed against a major B-cell epitope of the gp41 glycoprotein. When these antibodies bind the enzyme, the 3D conformation changes positively affecting the performance of the active site and, consequently, the enzyme activity is stimulated. Using 4-aminophenyl β-d-galactopyranoside (PAPG) as substrate yields p-aminophenol (PAP), which is reversibly oxidised at a very mild potential, ca. 0.37 V vs. Ag/AgCl over a range of electrode materials within the working pH range of β-galactosidase. In the present case, photolithographically produced microelectrode arrays resulted in a detection limit of 4 μM for 4-aminophenol (PAP). The presence of anti-HIV antibodies results in enzyme activity increases above 50% which, combined with the sensitivity and response time afforded by the microelectrode arrays, allowed for the diagnosis of HIV in sera samples within an hour.
Description6 pages, 5 figures, 1 table.-- PMID: 19393360 [PubMed].-- Printed version published May 8, 2009.
Publisher version (URL)http://dx.doi.org/10.1016/j.aca.2009.03.008
URIhttp://hdl.handle.net/10261/12262
DOI10.1016/j.aca.2009.03.008
ISSN0003-2670
Appears in Collections:(IMB-CNM) Artículos
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