Fast and easy enzyme immobilization by photoinitiated polymerization for efficient bioelectrochemical devices

Anal Chem. 2011 Apr 1;83(7):2824-8. doi: 10.1021/ac200297r. Epub 2011 Mar 15.


Immobilization and electrical wiring of enzymes is of particular importance for the elaboration of efficient biosensors and can be cumbersome. Here, we report a fast and easy protocol for enzyme immobilization, and as a proof of concept, we applied it to the immobilization of bilirubin oxidase, a labile enzyme. In the first step, bilirubin oxidase is mixed with a redox hydrogel "wiring" the enzyme reaction centers to electrodes. Then, this adduct is covered by an outer layer of PEGDA made by photoinitiated polymerization of poly(ethylene-glycol) diacrylate (PEGDA) and a photoclivable precursor, DAROCUR. This two-step protocol is 18 times faster than the current state-of-the-art protocol and leads to currents 25% higher. In addition, the outer layer of PEGDA acts as a protective layer increasing the lifetime of the electrode by 100% when operating continuously for 2000 s and by 60% when kept in dry state for 24 h. This new protocol is particularly appropriate for labile enzymes that quickly denaturate. In addition, by tuning the ratio PEGDA/DAROCUR, it is possible to make the enzyme electrodes even more active or more stable.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Electrochemistry / instrumentation*
  • Electrodes
  • Enzymes, Immobilized / chemistry*
  • Kinetics
  • Oxidoreductases Acting on CH-CH Group Donors / chemistry
  • Photochemical Processes*
  • Polyethylene Glycols / chemistry
  • Polymerization*
  • Polyporales / enzymology
  • Time Factors


  • Enzymes, Immobilized
  • poly(ethylene glycol)diacrylate
  • Polyethylene Glycols
  • Oxidoreductases Acting on CH-CH Group Donors
  • bilirubin oxidase