Ultraviolet photolithographic development of polyphosphazene hydrogel microstructures for potential use in microarray biosensors

Polyphosphazenes that bear both methoxyethoxyethoxy and cinnamyl side groups were synthesized and evaluated for use as hydrogels incorporated into micrometer-scale biosensor arrays. Polymers with the general formula [NPRxR'(y)](n) where R = OCH2CH2OCH2CH2OCH3 and R' = OCH=CHCH2Ph (x = y = 1; x = 2, y = 0) were synthesized. The polymers were cross-linked to form hydrogels by exposure to ultraviolet radiation (lambda = 320-480 nm) in the presence of a photoinitiator. Hydrogel microstructures in the size range 50-500 mu m were fabricated using standard photolithographic techniques. The resolution and dimensions of these microstructures were examined by optical microscopy, scanning electron microscopy, and profilometry. The resultant three-dimensional hydrogel microstructures were used to encapsulate enzymes for biosensor applications. The enzymatic activity of encapsulated horseradish peroxidase (HRP) was examined as a model system. The HRP catalyzed reaction between H2O2 and Amplex Red to produce a fluorescent product, resorufin, was confirmed by fluorescence microscopy.