Novel looped enzyme-polyamidoamine dendrimer nanohybrids used as biosensor matrix.

We report the synthesis of a novel looped enzyme-polyamidoamine nanocomposite with high enzyme loading density and long-term retention of bioactivity. The horseradish peroxidase (HRP) is first immobilized on fourth-grade (G4) poly(amidoamine) (PAMAM) dendrimer to form relatively a small enzyme-PAMAM composite, which is allowed to grow up into a larger one. The looped horseradish peroxidase-polyamidoamine (HRP-PAMAM) nanohybrid was characterized by TEM. The material obtained shows promising features as applied to the fabrication of high sensitive and long lifetime biosensors. In the presence of the hydroquinone mediator in the solution, the immobilized HRP exhibited excellent electro-catalytical response to H2O2. Under the optimal conditions, the resulting biosensor showed a linear response to H2O2 over a concentration range from 3.1x10(-6) to 2.0x10(-3) mol L(-1) with a sensitivity of 0.36 A L mol(-1) cm(-2) and a detection limit of 8.0x10(-7) mol L(-1). The sensitivity of the sensor response maintained over 70% of the original over 10 weeks. The catalytic activity of the looped enzyme-PAMAM nanohybrid form of HRP enzyme was obviously stabilized. As an extension, bienzyme sensor modified with glucose oxidase and HRP enzymatic PAMAM nanocomposites was constructed. The sensor exhibited improved performance and can be applied to the detection of glucose in real samples.

[1]  Younan Xia,et al.  Polyol Synthesis of Silver Nanoparticles: Use of Chloride and Oxygen to Promote the Formation of Single-Crystal, Truncated Cubes and Tetrahedrons , 2004 .

[2]  S. Moon,et al.  Improvement of an enzyme electrode by poly(vinyl alcohol) coating for amperometric measurement of phenol. , 2007, Talanta.

[3]  James R. Dewald,et al.  A New Class of Polymers: Starburst-Dendritic Macromolecules , 1985 .

[4]  Murali Sastry,et al.  Free-standing nanogold membranes as scaffolds for enzyme immobilization. , 2004, Langmuir : the ACS journal of surfaces and colloids.

[5]  A. Beezer,et al.  Dendrimers as potential drug carriers; encapsulation of acidic hydrophobes within water soluble PAMAM derivatives , 2003 .

[6]  Aiguo Wu,et al.  A method to construct a third-generation horseradish peroxidase biosensor: self-assembling gold nanoparticles to three-dimensional sol-gel network. , 2002, Analytical chemistry.

[7]  C. H. Walker,et al.  Synthesis and size control of gold nanoparticles stabilized by poly(methylphenylphosphazene). , 2001, Journal of the American Chemical Society.

[8]  Guo-Li Shen,et al.  Functionalization of multi-walled carbon nanotubes with poly(amidoamine) dendrimer for mediator-free glucose biosensor , 2007 .

[9]  Kim,et al.  Multilayered assembly of dendrimers with enzymes on gold: thickness-controlled biosensing interface , 2000, Analytical chemistry.

[10]  S. Iijima Helical microtubules of graphitic carbon , 1991, Nature.

[11]  Yuehe Lin,et al.  Enzyme nanoparticles-based electronic biosensor. , 2005, Chemical communications.

[12]  T. Hianik,et al.  Properties of glucose biosensors based on dendrimer layers. Effect of enzyme immobilization , 2002, Analytical and bioanalytical chemistry.

[13]  Jungbae Kim,et al.  Single-Enzyme Nanoparticles Armored by a Nanometer-Scale Organic/Inorganic Network , 2003 .

[14]  J. Luong,et al.  Electrochemical biosensing platforms using platinum nanoparticles and carbon nanotubes. , 2004, Analytical chemistry.

[15]  S. Yau,et al.  Preserved enzymatic activity of glucose oxidase immobilized on unmodified electrodes for glucose detection. , 2007, Biosensors & bioelectronics.

[16]  Lai-Kwan Chau,et al.  Amperometric L-lactate sensor based on sol-gel processing of an enzyme-linked silicon alkoxide. , 2007, Analytical chemistry.

[17]  Maogen Zhang,et al.  Carbon nanotube-chitosan system for electrochemical sensing based on dehydrogenase enzymes. , 2004, Analytical chemistry.

[18]  V. Rotello,et al.  Monolayer-controlled substrate selectivity using noncovalent enzyme-nanoparticle conjugates. , 2004, Journal of the American Chemical Society.

[19]  S. Cosnier,et al.  Sol-gel derived composite materials for the construction of oxidase/peroxidase mediatorless biosensors , 1997 .

[20]  G. Shen,et al.  A hydrogen peroxide biosensor based on nano-Au/PAMAM dendrimer/cystamine modified gold electrode , 2005 .

[21]  R. Yu,et al.  Amperometric glucose biosensor based on electrodeposition of platinum nanoparticles onto covalently immobilized carbon nanotube electrode. , 2007, Talanta.

[22]  M. Nath,et al.  New metal disulfide nanotubes. , 2001, Journal of the American Chemical Society.

[23]  Yuehe Lin,et al.  Solubilization of carbon nanotubes by Nafion toward the preparation of amperometric biosensors. , 2003, Journal of the American Chemical Society.

[24]  Michael B. Hall,et al.  Dendritic macromolecules: synthesis of starburst dendrimers , 1986 .

[25]  Xin Wei,et al.  Integration of enzymes and electrodes: spectroscopic and electrochemical studies of chitosan-enzyme films. , 2002, Analytical chemistry.

[26]  Yuehe Lin,et al.  Glucose Biosensors Based on Carbon Nanotube Nanoelectrode Ensembles , 2004 .