Fabrication of polymeric electron-transfer mediator/enzyme hydrogel multilayer on an Au electrode in a layer-by-layer process

Ziyin Li, Tomohiro Konno, Madoka Takai, Kazuhiko Ishihara

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30 Citations (Scopus)


The layer-by-layer (LBL) construction of an enzyme electrode covered with a multilayer structure alternately composed of a polymeric electron transfer mediator and a polymer-modified enzyme was examined. Poly(2-methacryloyloxyethyl phosphorylcholine- co- p-vinylphenylboronic acid- co-vinylferrocene) (PMVF) was synthesized and used as a polymeric electron transfer mediator. Glucose oxidase (GOx) was selected as a model enzyme and poly(vinyl alcohol) (PVA) chains were bound to the GOx (GOx-PVA) under mild conditions. The PMVF and PVA formed a gel spontaneously through a selective reaction between phenylboronic acid units and hydroxyl groups in both polymers. Using the spin coating technique, a repeating PMVF/GOx-PVA multilayer was fabricated on the surface of an Au electrode. The thickness of each PMVF/GOx-PVA layer was around 5.8. nm, corresponding to the dimensions of GOx. The electrochemical performance of the electrode was evaluated in glucose concentration measurement. The oxidation current of glucose by GOx was measured at 0.38. V (vs. Ag/AgCl), verifying that ferrocene units in the PMVF of the hydrogel electrically wired the immobilized GOx. Moreover, the current increased with the number of PMVF/GOx-PVA layers. That is, both intermolecular electron transfer between each individual layer and the presence of a freely diffusing substrate in the hydrogel were achieved. We conclude that a LBL structure constructed from PMVF and a PVA-modified enzyme is effective for use in developing bioelectronic devices that employ enzyme molecules.

Original languageEnglish
Pages (from-to)191-196
Number of pages6
JournalBiosensors and Bioelectronics
Issue number1
Publication statusPublished - 2012 Apr 15


  • 2-Methacryloyloxyethyl phosphorylcholine polymer
  • Electron transfer mediator
  • Enzymatic electrode
  • Hydrogel
  • Layer-by-layer


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