Biotechnology and Bioprocess Engineering

, Volume 21, Issue 4, pp 573–579 | Cite as

Immobilization of glucose oxidase on graphene oxide for highly sensitive biosensors

  • Sung-Gil Hong
  • Jae Hyun Kim
  • Ryang Eun Kim
  • Seok-Joon Kwon
  • Dae Woo Kim
  • Hee-Tae Jung
  • Jonathan S. Dordick
  • Jungbae Kim
Research Paper


Glucose oxidase (GOx) was immobilized onto graphene oxide (GRO) via three different preparation methods: enzyme adsorption (EA), enzyme adsorption and crosslinking (EAC), and enzyme adsorption, precipitation and crosslinking (EAPC). EAPC formulations, prepared via enzyme precipitation with 60% ammonium sulfate, showed 1,980 and 1,630 times higher activity per weight of GRO than those of EA and EAC formulations, respectively. After 59 days at room temperature, EAPC maintained 88% of initial activity, while EA and EAC retained 42 and 45% of their initial activities, respectively. These results indicate that the steps of precipitation and crosslinking in the EAPC formulation are critical to achieve high enzyme loading and stability of EAPC. EA, EAC and EAPC were used to prepare enzyme electrodes for use as glucose biosensors. Optimized EAPC electrode showed 93- and 25-fold higher sensitivity than EA and EAC, respectively. To further increase the sensitivity of EAPC electrode, multi-walled carbon nanotubes (MWCNTs) were mixed with EAPC for the preparation of enzyme electrode. Surprisingly, the EAPC electrode with additional 99.5 wt% MWCNTs showed 7,800-fold higher sensitivity than the EAPC electrode without MWCNT addition. Immobilization and stabilization of enzymes on GRO via the EAPC approach can be used for the development of highly sensitive biosensors as well as to achieve high enzyme loading and stability.


enzyme adsorption/precipitation/crosslinking glucose oxidase graphene oxide biosensors carbon nanotubes 


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Copyright information

© The Korean Society for Biotechnology and Bioengineering and Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Sung-Gil Hong
    • 1
    • 2
  • Jae Hyun Kim
    • 1
    • 2
  • Ryang Eun Kim
    • 1
    • 2
  • Seok-Joon Kwon
    • 3
  • Dae Woo Kim
    • 4
  • Hee-Tae Jung
    • 4
  • Jonathan S. Dordick
    • 3
  • Jungbae Kim
    • 1
    • 2
  1. 1.Department of Chemical and Biological EngineeringKorea UniversitySeoulKorea
  2. 2.Green SchoolKorea UniversitySeoulKorea
  3. 3.Department of Chemical and Biological Engineering, and Center for Biotechnology and Interdisciplinary StudiesRensselaer Polytechnic InstituteTroyUSA
  4. 4.Department of Chemical and Biomolecular Eng. (BK-21 plus) & KAIST Institute for Nano centuryKorea Advanced Institute of Science and TechnologyDaejeonKorea

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