Enzyme-Gelatin electrochemical biosensors: Scaling down

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Abstract

In this article we investigate the possibility of scaling down enzyme-gelatin modified electrodes by spin coating the enzyme-gelatin layer. Special attention is given to the electrochemical behavior of the selected enzymes inside the gelatin matrix. A glassy carbon electrode was used as a substrate to immobilize, in the first instance, horse heart cytochrome c (HHC) in a gelatin matrix. Both a drop dried and a spin coated layer was prepared. On scaling down, a transition from diffusion controlled reactions towards adsorption controlled reactions is observed. Compared to a drop dried electrode, a spin coated electrode showed a more stable electrochemical behavior. Next to HHC, we also incorporated catalase in a spin coated gelatin matrix immobilized on a glassy carbon electrode. By spincoating, highly uniform sub micrometer layers of biocompatible matrices can be constructed. A full electrochemical study and characterization of the modified surfaces has been carried out. It was clear that in the case of catalase, gluteraldehyde addition was needed to prevent leaking of the catalase from the gelatin matrix. © 2012 by the authors.

Figures

  • Figure 1. The current potential behaviour of a GelB|C (1), a drop dried HHC|GelB|C (2) and a spin coated HHC|GelB|C (3) electrode in a 10 mmol·L−1 HEPES pH 7 buffer solution with a scan rate of 50 mV·s−1.
  • Figure 2. Logarithmic plot between the peak current Ip and the scan rate for a spin coated HHC|GelB|GC electrode.
  • Figure 3. Five successive cyclic voltammograms obtained at a spin coated HHC|GelB|GC electrode in a 10 mmol·L−1 HEPES pH 7 buffer solution with a scan rate of 50 mV·s−1.
  • Figure 4. ATR-IR spectra of a spin coated GelB|C (black, 1), Cat|GelB|C (green, 2) and HHC|GelB|C (red, 3) electrode after background correction.
  • Figure 5. The current potential behavior of a spin coated GelB|C (1–2) and a spin coated Cat|GelB|C (3) electrode in the absence (1) and presence of 10.3 mmol·L−1 H2O2 (2–3) in a 10 mmol·L−1 HEPES pH 7 buffer solution with a scan rate of 50 mV·s−1. Curve 4 is the current potential behavior obtained after curve 3 and polishing of the electrode.
  • Figure 6. Optical profile measurement showing a gelatin layer thickness of 7 µm. The gelatin was removed locally using an Excimer laser.
  • Figure 7. (a,b) Scanning Electron Microscope (SEM) image of a Gel|C electrode; (c,d) SEM image of a Cat|Gel|C electrode; (e,f) SEM image of a Cat|Gel|C electrode which was soaked in a buffer solution for 24 h.

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CITATION STYLE

APA

De Wael, K., De Belder, S., Pilehvar, S., Van Steenberge, G., Herrebout, W., & Heering, H. A. (2012). Enzyme-Gelatin electrochemical biosensors: Scaling down. Biosensors, 2(1), 101–113. https://doi.org/10.3390/bios2010101

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