Issue 8, 2009
From the journal:

Analyst

Potentiometric enzyme immunoassay using miniaturized anion-selective electrodes for detection

Júlia Szűcs,a   Ernö Pretschb  and  Róbert E. Gyurcsányi*ac  

* Corresponding authors

a Department of Inorganic and Analytical Chemistry, Budapest University of Technology and Economics, Szt. Gellért tér 4, H-1111 Budapest, Hungary
E-mail: robertgy@mail.bme.hu

b Institute of Biogeochemistry and Pollutant Dynamics, ETH Zürich, CH-8092 Zürich, Switzerland

c Research Group of Technical Analytical Chemistry, Hungarian Academy of Sciences, Szt. Gellért tér 4, H-1111 Budapest, Hungary

Abstract

An enzyme-linked immunosorbent assay (ELISA) for prostate specific antigen (PSA) detection in human serum was developed based on the potentiometric detection of 6,8-difluoro-4-methylumbelliferone (DiFMU). The assays were carried out in anti-human PSA capture antibody modified microtiter plates (150 µL volume). After incubation in the PSA-containing serum samples, β-galactosidase-labeled PSA tracer antibody was added. The β-galactosidase label catalyzed the hydrolysis of 6,8-difluoro-4-methylumbelliferyl-β-D-galactopyranoside (DiFMUG) and the resulting DiFMU− anion was detected by potentiometric microelectrodes with anion-exchanger membrane. The selectivity of the anion-exchanger electrode is governed by the lipophilicity of the anions in the sample. Since DiFMU− is much more lipophilic (log P = 1.83) than any of the inorganic anions normally present in the working buffers and occurs in its anionic form at the physiological pH (pKa = 4.19), it was chosen as the species to be detected. The potentiometric ELISA-based method detects PSA in serum with a linear concentration range of 0.1–50 ng/mL. These results confirm the applicability of potentiometric detection in diagnostic PSA assays. Owing to simple methodology and low cost, potentiometric immunoassays seem to offer a feasible alternative to the development of in vitro diagnostic platforms.

Graphical abstract: Potentiometric enzyme immunoassay using miniaturized anion-selective electrodes for detection

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Article information

DOI
https://doi.org/10.1039/B904321G
Article type
Paper
Submitted
03 Mar 2009
Accepted
26 May 2009
First published
12 Jun 2009

Analyst, 2009,134, 1601-1607

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Potentiometric enzyme immunoassay using miniaturized anion-selective electrodes for detection

J. Szűcs, E. Pretsch and R. E. Gyurcsányi, Analyst, 2009, 134, 1601 DOI: 10.1039/B904321G

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