Nanostructured fluorescent particles for glucose sensing

Self-assembled thin films containing embedded enzymes and fluorescent indicators are being developed for use as highly specific glucose biosensors. The sensors are fabricated using electrostatic Layer-by-Layer (LBL) adsorption to create oxygen-sensitive (Ruthenium-based) layers, the fluorescent intensity of which responds to changes in local oxygen levels. Oxygen is consumed locally by the reaction between glucose oxidase (GOx) molecules and glucose. Latex particles serve as the templates for our sensors and fabrication is carried out through the alternate adsorption of multiple levels of {GOx/polycation} and {Ruthenium-polycation/polyanion} bilayers. Additional fluorescence layers as well as fluorescent latex are being considered as internal intensity references to allow ratiometric monitoring. Films adsorbed to the nanoparticle templates are being studied to understand the fundamental chemical and optical properties, including enzymatic activity, spectral shape and emission intensity. Enzymatic activity is retained and stability is improved after adsorption, and increased surface area afforded by the particles allows use of increased numbers of molecules. Fluorescence is also maintained, though blue shifts are observed in emission spectra, and indicator activity remains. In vitro characterization studies demonstrate the feasibility of the particles as glucose biosensors, and future work will aim to optimize the response for neural monitoring.

[1]  Helmuth Möhwald,et al.  Assembly, structural characterization, and thermal behavior of layer-by-layer deposited ultrathin films of poly(vinyl sulfate) and poly(allylamine) , 1993 .

[2]  Dongsik Yoo,et al.  Investigations of New Self-Assembled Multilayer Thin Films Based on Alternately Adsorbed Layers of Polyelectrolytes and Functional Dye Molecules , 1995 .

[3]  G. Coté,et al.  Glucose monitoring using implanted fluorescent microspheres , 2000, IEEE Engineering in Medicine and Biology Magazine.

[4]  N. Kotov,et al.  Layer-by-Layer Self-Assembly of Polyelectrolyte-Semiconductor Nanoparticle Composite Films , 1995 .

[5]  B. Vainshtein,et al.  Successive deposition of alternate layers of polyelectrolytes and a charged virus , 1994 .

[6]  Gero Decher,et al.  Fuzzy Nanoassemblies: Toward Layered Polymeric Multicomposites , 1997 .

[7]  G. Sukhorukov,et al.  Assembly of thin films by means of successive deposition of alternate layers of DNA and poly(allylamine) , 1993 .

[8]  Johannes Schmitt,et al.  New nanocomposite films for biosensors: layer-by-layer adsorbed films of polyelectrolytes, proteins or DNA , 1993 .

[9]  H. Clark,et al.  Optical nanosensors for chemical analysis inside single living cells. 1. Fabrication, characterization, and methods for intracellular delivery of PEBBLE sensors. , 1999, Analytical chemistry.

[10]  G. Hicks,et al.  The Enzyme Electrode , 1967, Nature.

[11]  Katsuhiko Ariga,et al.  Assembling Alternate Dye−Polyion Molecular Films by Electrostatic Layer-by-Layer Adsorption , 1997 .

[12]  G L Coté,et al.  A fluorescence-based glucose biosensor using concanavalin A and dextran encapsulated in a poly(ethylene glycol) hydrogel. , 1999, Analytical chemistry.

[13]  Soo‐Hyoung Lee,et al.  Thin Film Optical Sensors Employing Polyelectrolyte Assembly , 2000 .

[14]  H. Clark,et al.  Optical nanosensors for chemical analysis inside single living cells. 2. Sensors for pH and calcium and the intracellular application of PEBBLE sensors. , 1999, Analytical chemistry.

[15]  A. Revzin,et al.  Glucose and lactate biosensors based on redox polymer/oxidoreductase nanocomposite thin films. , 2000, Analytical chemistry.

[16]  G. Sauerbrey Verwendung von Schwingquarzen zur Wägung dünner Schichten und zur Mikrowägung , 1959 .

[17]  M. Philbert,et al.  Fluorescent nanosensors for intracellular chemical analysis: decyl methacrylate liquid polymer matrix and ion-exchange-based potassium PEBBLE sensors with real-time application to viable rat C6 glioma cells. , 2001, Analytical chemistry.

[18]  Katsuhiko Ariga,et al.  ASSEMBLY OF MULTICOMPONENT PROTEIN FILMS BY MEANS OF ELECTROSTATIC LAYER-BY-LAYER ADSORPTION , 1995 .

[19]  T. Kunitake,et al.  Formation of Ultrathin Polymer Layers on Solid Substrates by Means of Polymerization-Induced Epitaxy and Alternate Adsorption , 1996 .

[20]  L. Fellows,et al.  Enzyme packed bed system for the on-line measurement of glucose, glutamate, and lactate in brain microdialysate. , 1992, Analytical chemistry.

[21]  J. Hubbell,et al.  Poly(l-lysine)-g-Poly(ethylene glycol) Layers on Metal Oxide Surfaces: Attachment Mechanism and Effects of Polymer Architecture on Resistance to Protein Adsorption† , 2000 .

[22]  Thomas E. Mallouk,et al.  Layer-by-Layer Assembly of Intercalation Compounds and Heterostructures on Surfaces: Toward Molecular "Beaker" Epitaxy , 1994 .