Potentiometric enzyme immunoassay using miniaturized anion-selective electrodes for detection

Júlia Szcs, Ernö Pretsch, R. Gyurcsányi

Research output: Contribution to journalArticle

25 Citations (Scopus)

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 L 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.

Original languageEnglish
Pages (from-to)1601-1607
Number of pages7
JournalThe Analyst
Volume134
Issue number8
DOIs
Publication statusPublished - 2009

Fingerprint

immunoassay
Prostate-Specific Antigen
Antigens
antigen
Immunoenzyme Techniques
Anions
anion
Electrodes
electrode
Negative ions
Enzymes
enzyme
Assays
Hymecromone
Galactosidases
Immunosorbents
assay
serum
Ion exchangers
Antibodies

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy
  • Electrochemistry
  • Biochemistry
  • Environmental Chemistry

Cite this

Potentiometric enzyme immunoassay using miniaturized anion-selective electrodes for detection. / Szcs, Júlia; Pretsch, Ernö; Gyurcsányi, R.

In: The Analyst, Vol. 134, No. 8, 2009, p. 1601-1607.

Research output: Contribution to journalArticle

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