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Immobilization of glucose oxidase on graphene oxide for highly sensitive biosensors

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Abstract

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.

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Authors and Affiliations

  1. Department of Chemical and Biological Engineering, Korea University, Seoul, 02841, Korea

    Sung-Gil Hong, Jae Hyun Kim, Ryang Eun Kim & Jungbae Kim

  2. Green School, Korea University, Seoul, 08241, Korea

    Sung-Gil Hong, Jae Hyun Kim, Ryang Eun Kim & Jungbae Kim

  3. Department of Chemical and Biological Engineering, and Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, USA

    Seok-Joon Kwon & Jonathan S. Dordick

  4. Department of Chemical and Biomolecular Eng. (BK-21 plus) & KAIST Institute for Nano century, Korea Advanced Institute of Science and Technology, Daejeon, 34141, Korea

    Dae Woo Kim & Hee-Tae Jung

Authors
  1. Sung-Gil Hong
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  2. Jae Hyun Kim
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  3. Ryang Eun Kim
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  4. Seok-Joon Kwon
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  5. Dae Woo Kim
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  7. Jonathan S. Dordick
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Correspondence to Jonathan S. Dordick or Jungbae Kim.

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These authors contributed equally to this work.

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Hong, SG., Kim, J.H., Kim, R.E. et al. Immobilization of glucose oxidase on graphene oxide for highly sensitive biosensors. Biotechnol Bioproc E 21, 573–579 (2016). https://doi.org/10.1007/s12257-016-0373-4

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  • DOI: https://doi.org/10.1007/s12257-016-0373-4

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