Generation of mouse monoclonal antibody against (-)-epigallocatechin gallate.
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 beta-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.
Kuzuhara
Takashi
T
Laboratory of Biochemistry, Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Yamashiro-cho, Tokushima 770-8514, Japan. kuzuhara@ph.bunri-u.ac.jp
Kise
Daisuke
D
Shirakawa
Yayoi
Y
Sasada
Kinuko
K
Suganuma
Masami
M
Fujiki
Hirota
H
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002392:Catechin / Q000031:analogs & derivatives* / Q000276:immunology; D002459:Cell Fusion; D004247:DNA / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance; D013954:Thyroglobulin / Q000737:chemistry
D000911:Antibodies, Monoclonal; D002392:Catechin; D004247:DNA; D013954:Thyroglobulin; C057580:gallocatechol