Amperometric microcells for alkaline phosphatase assay

R. Gyurcsányi, Andrea Bereczki, G. Nagy, Michael R. Neuman, Ernö Lindner

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

To develop simple electrochemical immunoassays, a screen printed amperometric microcell with graphite working and Ag/AgCl reference electrodes was tested for the determination of alkaline phosphatase enzyme (ALP) and anti-humanIgG conjugated ALP (α-hIgG-ALP) activity in 5-10 μl samples. To ensure reproducible, steady state conditions, the working electrode surface was coated with mass-transport controlling hydrogel layer. The kinetic response curves of the hydrogel coated electrodes were linear. In addition, the hydrogel layer reduced the nonspecific adsorption of the α-hIgG-ALP conjugate on the working electrode surface. The measurements were made in the range of 2 ÷ 4000 mU ml-1 enzyme activities using ascorbic acid 2-phosphate (AAP) as the enzyme substrate. AAP is commercially available, non-toxic and has excellent stability. The sensitivity of the determinations was about 71% of the sensitivity which could be achieved using p-aminophenylphosphate (PAPP), a not easily accessible and unstable enzyme substrate. The experimentally determined kinetic parameters of the ALP enzyme catalyzed reactions were the same with the bare and hydrogel layer coated electrodes.

Original languageEnglish
Pages (from-to)235-240
Number of pages6
JournalThe Analyst
Volume127
Issue number2
DOIs
Publication statusPublished - 2002

Fingerprint

Phosphatases
phosphatase
Alkaline Phosphatase
Assays
Enzymes
Hydrogel
assay
enzyme
electrode
Hydrogels
Electrodes
Ascorbic acid
Enzyme activity
ascorbic acid
enzyme activity
Phosphates
phosphate
substrate
kinetics
Graphite

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Amperometric microcells for alkaline phosphatase assay. / Gyurcsányi, R.; Bereczki, Andrea; Nagy, G.; Neuman, Michael R.; Lindner, Ernö.

In: The Analyst, Vol. 127, No. 2, 2002, p. 235-240.

Research output: Contribution to journalArticle

Gyurcsányi, R, Bereczki, A, Nagy, G, Neuman, MR & Lindner, E 2002, 'Amperometric microcells for alkaline phosphatase assay', The Analyst, vol. 127, no. 2, pp. 235-240. https://doi.org/10.1039/b105308f
Gyurcsányi, R. ; Bereczki, Andrea ; Nagy, G. ; Neuman, Michael R. ; Lindner, Ernö. / Amperometric microcells for alkaline phosphatase assay. In: The Analyst. 2002 ; Vol. 127, No. 2. pp. 235-240.
@article{6fc63032cc4b4371bb1819c4de5b0a6c,
title = "Amperometric microcells for alkaline phosphatase assay",
abstract = "To develop simple electrochemical immunoassays, a screen printed amperometric microcell with graphite working and Ag/AgCl reference electrodes was tested for the determination of alkaline phosphatase enzyme (ALP) and anti-humanIgG conjugated ALP (α-hIgG-ALP) activity in 5-10 μl samples. To ensure reproducible, steady state conditions, the working electrode surface was coated with mass-transport controlling hydrogel layer. The kinetic response curves of the hydrogel coated electrodes were linear. In addition, the hydrogel layer reduced the nonspecific adsorption of the α-hIgG-ALP conjugate on the working electrode surface. The measurements were made in the range of 2 ÷ 4000 mU ml-1 enzyme activities using ascorbic acid 2-phosphate (AAP) as the enzyme substrate. AAP is commercially available, non-toxic and has excellent stability. The sensitivity of the determinations was about 71{\%} of the sensitivity which could be achieved using p-aminophenylphosphate (PAPP), a not easily accessible and unstable enzyme substrate. The experimentally determined kinetic parameters of the ALP enzyme catalyzed reactions were the same with the bare and hydrogel layer coated electrodes.",
author = "R. Gyurcs{\'a}nyi and Andrea Bereczki and G. Nagy and Neuman, {Michael R.} and Ern{\"o} Lindner",
year = "2002",
doi = "10.1039/b105308f",
language = "English",
volume = "127",
pages = "235--240",
journal = "The Analyst",
issn = "0003-2654",
publisher = "Royal Society of Chemistry",
number = "2",

}

TY - JOUR

T1 - Amperometric microcells for alkaline phosphatase assay

AU - Gyurcsányi, R.

AU - Bereczki, Andrea

AU - Nagy, G.

AU - Neuman, Michael R.

AU - Lindner, Ernö

PY - 2002

Y1 - 2002

N2 - To develop simple electrochemical immunoassays, a screen printed amperometric microcell with graphite working and Ag/AgCl reference electrodes was tested for the determination of alkaline phosphatase enzyme (ALP) and anti-humanIgG conjugated ALP (α-hIgG-ALP) activity in 5-10 μl samples. To ensure reproducible, steady state conditions, the working electrode surface was coated with mass-transport controlling hydrogel layer. The kinetic response curves of the hydrogel coated electrodes were linear. In addition, the hydrogel layer reduced the nonspecific adsorption of the α-hIgG-ALP conjugate on the working electrode surface. The measurements were made in the range of 2 ÷ 4000 mU ml-1 enzyme activities using ascorbic acid 2-phosphate (AAP) as the enzyme substrate. AAP is commercially available, non-toxic and has excellent stability. The sensitivity of the determinations was about 71% of the sensitivity which could be achieved using p-aminophenylphosphate (PAPP), a not easily accessible and unstable enzyme substrate. The experimentally determined kinetic parameters of the ALP enzyme catalyzed reactions were the same with the bare and hydrogel layer coated electrodes.

AB - To develop simple electrochemical immunoassays, a screen printed amperometric microcell with graphite working and Ag/AgCl reference electrodes was tested for the determination of alkaline phosphatase enzyme (ALP) and anti-humanIgG conjugated ALP (α-hIgG-ALP) activity in 5-10 μl samples. To ensure reproducible, steady state conditions, the working electrode surface was coated with mass-transport controlling hydrogel layer. The kinetic response curves of the hydrogel coated electrodes were linear. In addition, the hydrogel layer reduced the nonspecific adsorption of the α-hIgG-ALP conjugate on the working electrode surface. The measurements were made in the range of 2 ÷ 4000 mU ml-1 enzyme activities using ascorbic acid 2-phosphate (AAP) as the enzyme substrate. AAP is commercially available, non-toxic and has excellent stability. The sensitivity of the determinations was about 71% of the sensitivity which could be achieved using p-aminophenylphosphate (PAPP), a not easily accessible and unstable enzyme substrate. The experimentally determined kinetic parameters of the ALP enzyme catalyzed reactions were the same with the bare and hydrogel layer coated electrodes.

UR - http://www.scopus.com/inward/record.url?scp=0036181692&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036181692&partnerID=8YFLogxK

U2 - 10.1039/b105308f

DO - 10.1039/b105308f

M3 - Article

VL - 127

SP - 235

EP - 240

JO - The Analyst

JF - The Analyst

SN - 0003-2654

IS - 2

ER -