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

J Zeravik, Tautgirdas Ruzgas, M Franek

Research output: Contribution to journalArticlepeer-review

31 Citations (SciVal)

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.
Original languageEnglish
Pages (from-to)1321-1327
JournalBiosensors & Bioelectronics
Volume18
Issue number11
DOIs
Publication statusPublished - 2003

Bibliographical note

The information about affiliations in this record was updated in December 2015.
The record was previously connected to the following departments: Analytical Chemistry (S/LTH) (011001004)

Subject classification (UKÄ)

  • Analytical Chemistry

Keywords

  • glucose oxidase
  • recombinant peroxidase
  • co-immobilization
  • antibody
  • enzyme-channeling
  • immunosensor

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