Mediatorless amperometric glucose biosensing using 3-aminopropyltriethoxysilane-functionalized graphene.

A mediatorless glucose biosensor was developed by the immobilization of glucose oxidase (GOx) to graphene-functionalized glassy carbon electrode (GCE). The surface of GCE was functionalized with graphene by incubating it with graphene dispersed in 3-aminopropyltriethoxysilane (APTES), which acted both as a dispersion agent for graphene and as an amine surface modification agent for GCE and graphene. This was followed by the covalent binding of GOx to graphene-functionalized GCE using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC) based crosslinking. Graphene provided signal enhancement by providing greater surface area for GOx binding, while APTES-functionalization led to a higher GOx immobilization density by providing free amino groups for crosslinking. The developed biosensor used a redox potential of -0.45 V (vs. Ag/AgCl) for detecting glucose in the diabetic pathophysiological range 0.5-32 mM. There was no interference from endogenous electroactive substances and drug metabolites. The developed biosensor was further validated for detecting blood glucose in commercial artificial blood glucose linearity standards in the range 1.4-27.9 mM. Therefore, it is ideal for diabetic blood glucose monitoring. The developed bioanalytical procedure for preparation of GOx-bound graphene-functionalized GCEs had high production reproducibility and high storage stability, which is appropriate for the commercial mass production of enzyme-bound electrodes.

[1]  Song Zhang,et al.  Bio-electrocatalysis of NADH and ethanol based on graphene sheets modified electrodes. , 2011, Talanta.

[2]  Xin Wang,et al.  Graphene−Metal Particle Nanocomposites , 2008 .

[3]  Jun Liu,et al.  Glucose biosensor based on immobilization of glucose oxidase in platinum nanoparticles/graphene/chitosan nanocomposite film. , 2009, Talanta.

[4]  Huafeng Yang,et al.  Direct electrochemistry of glucose oxidase and biosensing for glucose based on graphene. , 2009, Analytical chemistry.

[5]  Gregory P. Nordin,et al.  Single-sided inkjet functionalization of silicon photonic microcantilevers , 2012 .

[6]  Arben Merkoçi,et al.  Configurations used in the design of screen-printed enzymatic biosensors. A review , 2000 .

[7]  Anthony Turner,et al.  On the use of screen- and ink-jet printing to produce amperometric enzyme electrodes for lactate☆ , 1996 .

[8]  S. Stankovich,et al.  Graphene-based composite materials , 2006, Nature.

[9]  I. Karube,et al.  Direct electron transfer with glucose oxidase immobilized in an electropolymerized poly( N-methylpyrrole) film on a gold microelectrode , 1990 .

[10]  Chang Ming Li,et al.  Thin-walled graphitic nanocages as a unique platform for amperometric glucose biosensor. , 2010, ACS applied materials & interfaces.

[11]  S. K. Vashist,et al.  Effect of antibody immobilization strategies on the analytical performance of a surface plasmon resonance-based immunoassay. , 2011, The Analyst.

[12]  L. Setti,et al.  An amperometric glucose biosensor prototype fabricated by thermal inkjet printing. , 2005, Biosensors & bioelectronics.

[13]  Priscilla Kailian Ang,et al.  Solution-gated epitaxial graphene as pH sensor. , 2008, Journal of the American Chemical Society.

[14]  A. Govindaraj,et al.  Graphene-based electrochemical supercapacitors , 2008 .

[15]  G J Kost,et al.  Effects of drugs on glucose measurements with handheld glucose meters and a portable glucose analyzer. , 2000, American journal of clinical pathology.

[16]  T. Huang,et al.  Aminopropyltriethoxysilane (APTES)-functionalized nanoporous polymeric gratings: fabrication and application in biosensing , 2007 .

[17]  J. Luong,et al.  Carbocatalytic dehydration of xylose to furfural in water , 2012 .

[18]  Sandeep Kumar Vashist,et al.  Development of a high sensitivity rapid sandwich ELISA procedure and its comparison with the conventional approach. , 2010, Analytical chemistry.

[19]  Lutz Heinemann,et al.  Quality of glucose measurement with blood glucose meters at the point-of-care: relevance of interfering factors. , 2010, Diabetes technology & therapeutics.

[20]  Ping Wu,et al.  Detection of glucose based on direct electron transfer reaction of glucose oxidase immobilized on highly ordered polyaniline nanotubes. , 2009, Analytical chemistry.

[21]  S. K. Vashist,et al.  Evaluation of apparent non-specific protein loss due to adsorption on sample tube surfaces and/or altered immunogenicity. , 2011, The Analyst.

[22]  P. Desmeules,et al.  Disinfectant wipes containing hydrogen peroxide induce overestimation of glucose results obtained with Lifescan SureStep Flexx® glucose meter. , 2010, Clinical biochemistry.

[23]  W. Ryan,et al.  Whole Blood Glucose Standard is Key to Accurate Insulin Dosages , 2007, Journal of diabetes science and technology.

[24]  M. Lyon,et al.  Interference studies with two hospital-grade and two home-grade glucose meters. , 2009, Diabetes technology & therapeutics.

[25]  K. Novoselov,et al.  Detection of individual gas molecules adsorbed on graphene. , 2006, Nature materials.

[26]  F. M. Peeters,et al.  Adsorption of H 2 O , N H 3 , CO, N O 2 , and NO on graphene: A first-principles study , 2007, 0710.1757.

[27]  Jun Liu,et al.  Glucose oxidase-graphene-chitosan modified electrode for direct electrochemistry and glucose sensing. , 2009, Biosensors & bioelectronics.

[28]  Hui Zhang,et al.  Direct electrochemistry of glucose oxidase assembled on graphene and application to glucose detection , 2010 .

[29]  S. Dong,et al.  Electrochemical sensing and biosensing platform based on chemically reduced graphene oxide. , 2009, Analytical chemistry.

[30]  K. Müllen,et al.  Transparent, conductive graphene electrodes for dye-sensitized solar cells. , 2008, Nano letters.

[31]  Kun Wang,et al.  Enhanced direct electrochemistry of glucose oxidase and biosensing for glucose via synergy effect of graphene and CdS nanocrystals. , 2011, Biosensors & bioelectronics.

[32]  Chang Ming Li,et al.  Direct electron transfer of glucose oxidase and biosensing of glucose on hollow sphere-nanostructured conducting polymer/metal oxide composite. , 2010, Physical chemistry chemical physics : PCCP.

[33]  Chang Ming Li,et al.  Highly Sensitive Nitric Oxide Sensing Using Three‐Dimensional Graphene/Ionic Liquid Nanocomposite , 2011 .

[34]  Ying Wang,et al.  Application of graphene-modified electrode for selective detection of dopamine , 2009 .

[35]  Andre K. Geim,et al.  The rise of graphene. , 2007, Nature materials.

[36]  B. Pejcic,et al.  Functionalized graphene as an aqueous phase chemiresistor sensing material , 2011 .

[37]  Andre K. Geim,et al.  Electric Field Effect in Atomically Thin Carbon Films , 2004, Science.

[38]  Jun‐Jie Zhu,et al.  Fabrication of gold nanoparticles on bilayer graphene for glucose electrochemical biosensing , 2011 .

[39]  Xuping Sun,et al.  Synthesis of functional SiO₂-coated graphene oxide nanosheets decorated with Ag nanoparticles for H₂O₂ and glucose detection. , 2011, Biosensors & bioelectronics.

[40]  Freddy Yin Chiang Boey,et al.  Direct Electrochemical Reduction of Single-Layer Graphene Oxide and Subsequent Functionalization with Glucose Oxidase , 2009 .

[41]  Sandeep Kumar Vashist,et al.  Technology behind commercial devices for blood glucose monitoring in diabetes management: a review. , 2011, Analytica chimica acta.

[42]  Sandeep Kumar Vashist,et al.  Multisubstrate-compatible ELISA procedures for rapid and high-sensitivity immunoassays , 2011, Nature Protocols.

[43]  S. Stankovich,et al.  Synthesis and exfoliation of isocyanate-treated graphene oxide nanoplatelets , 2006 .

[44]  Jian-hui Jiang,et al.  Palladium nanoparticle/chitosan-grafted graphene nanocomposites for construction of a glucose biosensor. , 2011, Biosensors & bioelectronics.