We report a novel biosensor platform based on surface plasmon-enhanced fluorescence spectroscopy (SPFS) and a
responsive N-isopropylacrylamide (NIPAAm) hydrogel binding matrix. This binding matrix highly swells in aqueous
environment and it can be modified with receptor biomolecules by using active ester coupling chemistry. After the
binding of target analyte molecules contained in a sample by receptor biomolecules immobilized in the hydrogel matrix,
the captured analyte molecules can be compacted on the surface through the collapse of the gel triggered by an external
stimulus. A thin hydrogel NIPAAm-based film was attached to a gold sensor surface and modified with mouse IgG
receptor molecules. The affinity binding of antibodies against mouse IgG that were labeled with Alexa Fluor
chromophores was observed by surface plasmon-enhanced fluorescence spectroscopy. We demonstrate that the collapse
of the hydrogel matrix results in the enhancement of measured fluorescence intensity owing to the increase in the
concentration of captured molecules within the evanescent field of surface plasmons.
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