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Surface plasmon resonance-based trace detection of small molecules by competitive and signal enhancement immunoreaction |
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| Authors: | Aizawa Hidenobu Tozuka Mitsuhiro Kurosawa Shigeru Kobayashi Koichi Reddy Subrayal M Higuchi Masahiro |
| Affiliation: | a National Institute of Advanced Industrial Science & Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8565, Japan b Institute of Medical Science, Musashi Institute of Technology, 1-28-1 Tamatsutumi, Setagaya-ku, Tokyo 158-8557, Japan c School of Biomedical and Molecular Sciences, University of Surrey, Guildford, Surrey GU2 7XH, UK d Faculty of Engineering, Mie University, 1577 Kurimamachiya-cho, Tsu, Mie 514-8507, Japan |
| Abstract: | A surface plasmon resonance (SPR)-immunosensor for detection of the low molecular weight compound 2,4-dinitorophenol (DNP) at ultra-low concentration has been developed. The sensor strategy is based on a competitive immunoreaction between DNP and a DNP-protein conjugate, namely DNP-bovine serum albumin conjugate (DNP-BSA). Anti-DNP monoclonal antibody was immobilized on a gold thin-film coated SPR-sensor chip by means of a chemical coupling process. DNP-BSA, on contact with the anti-DNP antibody immobilized SPR-immunosensor chip causes an increase in the resonance angle of the sensor chip. The optimum concentration of immobilized antibody on the SPR-sensor chip is 100 μg mL−1. The SPR-immunosensor response for free DNP determination using the competitive immunoreaction had a response time of ca. 15 min. Using this method, DNP could be determined in the concentration range 1 ppt to 1 ppb. The SPR signal for ppt levels of DNP was enhanced by a factor of three by subsequently treating immuno-bound DNP-BSA with a secondary anti-DNP antibody. |
| Keywords: | Surface plasmon resonance (SPR) SPR-immunosensor 2,4-Dinitrophenol (DNP) DNP-protein conjugate Anti-DNP-protein monoclonal antibody Signal enhancement immunoreaction |
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