Analytical Sciences

Competitive ELISA-like Label-free Detection of Lysozyme by Using a Fluorescent Monomer-doped Molecularly Imprinted Hydrogel

Takuya KUBO,* Shunsuke ARIMURA,* Toyohiro NAITO,* Tomoharu SANO,** and Koji OTSUKA*

*Graduate School of Engineering, Kyoto University, Katsura, Nishikyo, Kyoto 615-8510, Japan
**Center for Environmental Measurement and Analysis, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki 305-8506, Japan

We report on a molecularly imprinted hydrogel for the selective fluorescent detection of a targeting protein. To achieve the fluorescent detection of proteins, a fluorescent functional monomer was synthesized with fluorescein isothiocyanate and allylamine. The monomer (FITC-AA) provided a gradable alteration of the absorption spectrum, and a decrease of the fluorescent intensity due to an interaction with the model protein, lysozyme. Also, the imprinted hydrogel (PI-gel) was simply prepared with FITC-AA, the anionic functional monomers, poly(ethylene glycol) dimethacrylate, and lysozyme by radical photo polymerization. The PI-gel showed selective adsorption for lysozyme and the imprinting factor reached to 2.7. Additionally, the PI-gel indicated a gradual decrease of the fluorescent intensity corresponding to the lysozyme concertation similar to a competitive enzyme-linked immunosorbent assay detection.

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