Generation of Mouse Monoclonal Antibody against (-)-Epigallocatechin Gallate

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  • Kuzuhara Takashi
    Laboratory of Biochemistry, Faculty of Pharmaceutical Sciences, Tokushima Bunri University
  • Kise Daisuke
    Laboratory of Biochemistry, Faculty of Pharmaceutical Sciences, Tokushima Bunri University
  • Shirakawa Yayoi
    Laboratory of Biochemistry, Faculty of Pharmaceutical Sciences, Tokushima Bunri University
  • Sasada Kinuko
    Laboratory of Biochemistry, Faculty of Pharmaceutical Sciences, Tokushima Bunri University
  • Suganuma Masami
    Research Institute for Clinical Oncology, Saitama Cancer Center
  • Fujiki Hirota
    Laboratory of Biochemistry, Faculty of Pharmaceutical Sciences, Tokushima Bunri University

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Abstract

Green tea is an acknowledged cancer preventive in Japan, and the main constituent of green tea catechins is (−)-epigallocatechin gallate (EGCG). To investigate the bioavailability of EGCG in humans, we generated a monoclonal antibody against EGCG in BALB/c mice by immunizing thyroglobulin-conjugated EGCG. Out of 32 hybridoma cell lines, three hybridomas were selected by enzyme-linked immunosorbent assay (ELISA), and then determined by surface plasmon resonance assay: One hybridoma TG38 produced a specific monoclonal antibody against EGCG. The primary structure of TG38 light chain was then deduced from DNA sequence of the light chain gene. The NCBI-BLAST search showed the uniqueness of TG38 monoclonal antibody, and three amino acid residues specific for TG38 were aligned on two loops and one β-sheet of the tertiary structure of the antibody. The TG38 antibody is the first monoclonal antibody against EGCG and catechins, since it bound to four green tea catechins with a galloyl group.

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