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A highly sensitive flow-through amperometric immunosensor based on the peroxidase chip and enzyme-channeling principle

Zeravik, J; Ruzgas, Tautgirdas LU and Franek, M (2003) In Biosensors & Bioelectronics 18(11). p.1321-1327
Abstract
A concept based on the Peroxidase-chip (P-chip), antibody co-immobilization, competitive and enzyme-channeling principle was exploited to develop an integrated flow-through amperometric biosensor for detection of environmental pollutants such as s-triazine herbicides. In this concept, recombinant peroxidase is immobilized on the gold electrode (P-chip) in such a way that direct electron transfer is achieved. The recognition and quantitation the target analyte is realized through the competition between the simazine-glucose oxidase (GOD) conjugate and free simazine for the binding sites of the monoclonal antibody co-immobilized with peroxidase on the gold electrode. The arrangement allows to generate a specific signal in the presence of... (More)
A concept based on the Peroxidase-chip (P-chip), antibody co-immobilization, competitive and enzyme-channeling principle was exploited to develop an integrated flow-through amperometric biosensor for detection of environmental pollutants such as s-triazine herbicides. In this concept, recombinant peroxidase is immobilized on the gold electrode (P-chip) in such a way that direct electron transfer is achieved. The recognition and quantitation the target analyte is realized through the competition between the simazine-glucose oxidase (GOD) conjugate and free simazine for the binding sites of the monoclonal antibody co-immobilized with peroxidase on the gold electrode. The arrangement allows to generate a specific signal in the presence of glucose through the channeling of H2O2 produced by GOD conjugate bound to the antibody. The immunosensor exhibited 50% signal decrease (IC50 value) at aproximately 0.02 mug l(-1). A concentration of 0.1 ng l(-1) gave a signal clearly distinguishable from the blank whereas the ELISA using the same antibody had a typical detection limit of about 1 mug l(-1), which is four orders of magnitude higher compared to the presented biosensor system. The results demonstrated that gene engineering biomolecules, in this case recombinant peroxidase, might be attractive reagents for the development of electrochemical immunosensors. (C) 2003 Elsevier Science B.V. All rights reserved. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
glucose oxidase, recombinant peroxidase, co-immobilization, antibody, enzyme-channeling, immunosensor
in
Biosensors & Bioelectronics
volume
18
issue
11
pages
1321 - 1327
publisher
Elsevier
external identifiers
  • pmid:12896832
  • wos:000185193500002
  • scopus:0042706515
ISSN
1873-4235
DOI
10.1016/S0956-5663(03)00076-9
language
English
LU publication?
yes
id
7c09dab4-0679-4a10-8266-a96428930ca4 (old id 301781)
date added to LUP
2007-09-24 15:28:27
date last changed
2018-01-07 09:37:27
@article{7c09dab4-0679-4a10-8266-a96428930ca4,
  abstract     = {A concept based on the Peroxidase-chip (P-chip), antibody co-immobilization, competitive and enzyme-channeling principle was exploited to develop an integrated flow-through amperometric biosensor for detection of environmental pollutants such as s-triazine herbicides. In this concept, recombinant peroxidase is immobilized on the gold electrode (P-chip) in such a way that direct electron transfer is achieved. The recognition and quantitation the target analyte is realized through the competition between the simazine-glucose oxidase (GOD) conjugate and free simazine for the binding sites of the monoclonal antibody co-immobilized with peroxidase on the gold electrode. The arrangement allows to generate a specific signal in the presence of glucose through the channeling of H2O2 produced by GOD conjugate bound to the antibody. The immunosensor exhibited 50% signal decrease (IC50 value) at aproximately 0.02 mug l(-1). A concentration of 0.1 ng l(-1) gave a signal clearly distinguishable from the blank whereas the ELISA using the same antibody had a typical detection limit of about 1 mug l(-1), which is four orders of magnitude higher compared to the presented biosensor system. The results demonstrated that gene engineering biomolecules, in this case recombinant peroxidase, might be attractive reagents for the development of electrochemical immunosensors. (C) 2003 Elsevier Science B.V. All rights reserved.},
  author       = {Zeravik, J and Ruzgas, Tautgirdas and Franek, M},
  issn         = {1873-4235},
  keyword      = {glucose oxidase,recombinant peroxidase,co-immobilization,antibody,enzyme-channeling,immunosensor},
  language     = {eng},
  number       = {11},
  pages        = {1321--1327},
  publisher    = {Elsevier},
  series       = {Biosensors & Bioelectronics},
  title        = {A highly sensitive flow-through amperometric immunosensor based on the peroxidase chip and enzyme-channeling principle},
  url          = {http://dx.doi.org/10.1016/S0956-5663(03)00076-9},
  volume       = {18},
  year         = {2003},
}