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  • Development of 3-hydroxybutyrate dehydrogenase enzyme biosensor based on carbon nanotube-modified screen-printed electrode.

Development of 3-hydroxybutyrate dehydrogenase enzyme biosensor based on carbon nanotube-modified screen-printed electrode.

IET nanobiotechnology (2013-05-28)
Fahimeh Khorsand, Siavash Riahi, Solmaz Eynollahi Fard, Soheila Kashanian, Ali Naeemy, Bagher Larijani, Kobra Omidfar
ABSTRACT

Precise detection of 3-hydroxybutyrate (HB) in biological samples is of great importance for management of diabetic patients. In this study, an HB biosensor based on single-walled carbon nanotubes (SWCNTs)-modified screen-printed electrode (SPE) was developed to determine the concentration of HB in serum. The specific detecting enzyme, HB dehydrogenase, was physically immobilised on SWCNTs deposited on the surface of SPEs. The electrochemical measurement of HB that involved cyclic voltammetry was based on the sAgnal produced by j3-nicotinamide adenine dinucleotide (NADH), one of the products of the enzymatic reaction. The application of SWCNT reduced the oxidation potential of NADH to about -0.05 V. Electrochemical measurements showed that the response of this biosensor had relevant good linearity in the range of 0.1-2 mM with a low detection limit of 0.009 mM. Investigation of biosensor response in the presence of interfering molecules verified its specificity. Furthermore, the study of long-term stability demonstrated the acceptable efficiency of this biosensor for about 100 days.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
β-Hydroxybutyrate Dehydrogenase from Rhodopseudomonas sphaeroides, Type V, lyophilized powder, 250-750 units/mg protein
Sigma-Aldrich
3-Hydroxybutyric acid, 95%
Roche
3-Hydroxybutyrate Dehydrogenase (3-HBDH), suspension, ~3 units/mg protein (At 25 °C with 3-hydroxybutyrate as the substrate; standardized with BSA.), grade II
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide, pkg of 10 mg (per vial)
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide, pkg of 50 mg (per vial)
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide, pkg of 20 mg (per vial)
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide hydrate, ≥95% (HPLC)
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide hydrate, ≥99%
Sigma-Aldrich
β-Hydroxybutyrate Dehydrogenase from Pseudomonas lemoignei, lyophilized powder, ≥200 units/mg protein
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide hydrate, suitable for cell culture, ≥96.5% (HPLC), ≥96.5% (spectrophotometric assay), from yeast
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide hydrate, ≥96.5% (HPLC), ≥96.5% (spectrophotometric assay), from yeast
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide hydrate, BioUltra, ≥98%, from yeast
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide hydrate, Grade AA-1
Sigma-Aldrich
β-Nicotinamide adenine dinucleotide hydrate, purified by column chromatography, ≥99%