Issue 6, 2012

Magnetic microbead-based enzyme immunoassay for ovalbumin using hydrodynamic voltammetry and fluorometric detection

Abstract

A paramagnetic microbead-based enzyme immunoassay was demonstrated for detecting ovalbumin (OVA). The immunoassay sandwich was made by attaching a biotinylated antibody to the streptavidin coated beads as a mobile solid phase, capturing antigen, and then exposing the antigen to an antibody conjugated with β-galactosidase. β-Galactosidase converts p-aminophenyl galactopyranoside (PAPG) and fluorescein di-β-D-galactopyranoside (FDG) to p-aminophenol (PAP) and fluorescein, respectively. The current response of PAP generated by the enzymatic reaction of β-galactosidase was detected with hydrodynamic voltammetry in a droplet using a rotating disk electrode (RDE) system. The performance of this electrochemical assay was compared with fluorometric detection of fluorescein produced by the same assay system. The limits of detection for OVA determined by hydrodynamic amperometry and fluorometry were 2.1 nM (43 fmol) and 2.9 nM (58 fmol), respectively. Furthermore, the effects of the conditions commonly found in drinking water supply systems on the OVA assay were also evaluated.

Graphical abstract: Magnetic microbead-based enzyme immunoassay for ovalbumin using hydrodynamic voltammetry and fluorometric detection

Article information

Article type
Paper
Submitted
29 Oct 2011
Accepted
28 Mar 2012
First published
04 May 2012

Anal. Methods, 2012,4, 1783-1789

Magnetic microbead-based enzyme immunoassay for ovalbumin using hydrodynamic voltammetry and fluorometric detection

H. Kuramitz, H. B. Halsall and W. R. Heineman, Anal. Methods, 2012, 4, 1783 DOI: 10.1039/C2AY05738G

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