Analytical Sciences

Detection of Mast Cells Expressing c-Kit Using Antibody Covalently Bound to Gelatin Elongated from Surface of Immunosensor Based on Surface Plasmon Resonance

Shiro MIYAKE,*,** Daisuke IRIKURA,** and Tomomi YAMASAKI***

*Department of Food and Life Science, Azabu University, 1-17-71 Fuchinobe, Chuo, Sagamihara, Kanagawa 252-5201, Japan
**Research & Development Division, Horiba, Ltd., 2 Miyanohigashicho, Kisshoin, Minami, Kyoto 601-8510, Japan
***Osaka Institute of Public Health, 1-3-69 Nakamichi, Higashinari, Osaka 537-0025, Japan

An immunosensor based on surface plasmon resonance was applied to detect mast cells expressing c-Kit. Sufficient detection of the mast cells was achieved by covalent immobilization of gelatin firstly on the sensor surface and followed by covalent binding of the anti-c-Kit antibody to lysine residues in the gelatin molecules through bis(sulfosuccinimidyl)suberate (BS3) treatment. By using BS3, which is a homo-bifunctional reagent, the lysine residues of the anti-c-Kit antibody easily bound to the lysine residues of the gelatin in the physiological condition. The lower limit of detection was 10⁴ cells/mL.

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