A bilirubin biosensor based on a multilayer network enzyme electrode

Benjamin Shoham, Yoelit Migron, Azalia Riklin, Itamar Willner*, Boris Tartakovsky

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

95 Scopus citations

Abstract

This paper describes the construction of a bilirubin oxidase multilayer electrode that responds amperometrically to bilirubin concentration. The enzyme electrode was constructed by the covalent attachment of bilirubin oxidase layers to a self-assembled monolayer of 3-mercaptopropionate ester, which was associated with a gold surface. Electron transfer from the enzyme redox site to the electrode was mediated by ferrocene carboxylic acid. The multilayer electrode responded linearly to bilirubin concentration in solution. In contrast to the native enzyme, which is unstable, the bilirubin oxidase multilayer electrode was stable, and its shelf-life was more than three months at 4°C. The amperometric response of the enzyme electrode towards bilirubin was dependent on the albumin concentration in the system. By adjustment of this concentration to the value present in newborns' serum, a calibration curve for the amperometric response as a function of bilirubin was deduced. The concentration of bilirubin in serum samples from newborns was determined.

Original languageEnglish
Pages (from-to)341-352
Number of pages12
JournalBiosensors and Bioelectronics
Volume10
Issue number3-4
DOIs
StatePublished - 1995

Keywords

  • Amperometric biosensor
  • bilirubin
  • bilirubine oxidase
  • biosensor
  • electron transfer in proteins
  • enzyme electrode
  • enzyme monolayer
  • enzyme multilayers

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