Glucose and Lactate Biosensors Based on Redox Polymer/Oxidoreductase Nanocomposite Thin Films

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Abstract
Glucose and lactate enzyme electrodes have been fabricated through the deposition of an anionic self-assembled monolayer and subsequent redox polymer/enzyme electrostatic complexation on gold substrates. These surfaces were functionalized with a negative charge using 11-mercaptoundecanoic acid (MUA), followed by alternating immersions in cationic redox polymer solutions and anionic glucose oxidase (GOX) or lactate oxidase (LAX) solutions to build the nanocomposite structure. The presence of the multilayer structure was verified by ellipsometry and sensor function characterized electrochemically. Reproducible analyte response curves from 2 to 20 mM (GOX) and 2−10 mM (LAX) were generated with the standard deviation between multiple sensors between 12 and 17%, a direct result of the reproducibility of the fabrication technique. In the case of glucose enzyme electrodes, the multilayer structure was further stabilized through the introduction of covalent bonds within and between the layers. Chemical cross-linking was accomplished by exposing the thin film to glutaraldehyde vapors, inducing linkage formation between lysine and arginine residues present on the enzyme periphery with amine groups present on a novel redox polymer, poly[vinylpyridine Os(bisbipyridine)2Cl]-co-allylamine. Finally, an initial demonstration of thin-film patterning was performed as a precursor to the development of redundant sensor arrays. Microcontact printing was used to functionalize portions of a gold surface with a blocking agent, typically 1-hexadecanethiol. This was followed by immersion in MUA to functionalize the remaining portions of gold with negative charges. The multilayer deposition process was then followed, resulting in growth only on the regions containing MUA, resulting in a “positive”-type pattern. This technique may be used for fabrication of thin-film redundant sensor arrays, with thickness under 100 Å and lateral dimensions on a micrometer scale.

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