title: Epitope mapping of monoclonal antibodies for the Deinococcus radiodurans bacteriophytochome.
abstract: Bacteriophytochromes (BphP) are phytochrome-like light sensing proteins in bacteria, which use biliverdin as a chromophore. In order to study the biochemical properties of the DrBphP protein, five (2B8, 2C11, 3B2, 3D2, and 3H7) anti-DrBphP monoclonal antibodies were produced through the immunization of mice with purified full-length DrBphP and DrBphN (1-321 amino acid) proteins, and epitope mapping was then carried out. Among the five antibodies, 2B8 and 2C11 preferentially recognized the N-terminal region of BphP whereas 3B2, 3D2, and 3H7 showed preference for the C-terminal region. We performed further epitope mapping using recombinant truncated BphP proteins to narrow down their target sequences. The results demonstrated that each of the five monoclonal antibodies recognized different regions on the DrBphP protein. Additionally, epitopes of 2B8 and 3H7 antibodies were discovered to be shorter than 10 amino acids (2B8: RDPLPFFPP, 3H7: PGEIEEA). These two antibodies with such specific recognition epitopes could be especially valuable for developing new peptide tags for protein detection and purification.
authors.affiliation: Graduate School of Biotechnology and Plant Metabolism Research Center, Kyung Hee University, Yongin, 446-701, Korea.
authors.forename: Tae-Lim
authors.initials: TL
authors.lastname: Kim
authors.forename: Jihey
authors.initials: J
authors.lastname: Yoo
authors.forename: Kanidta
authors.initials: K
authors.lastname: Sangsawang
authors.forename: Man-Ho
authors.initials: MH
authors.lastname: Cho
authors.forename: Seung Hwan
authors.initials: SH
authors.lastname: Yang
authors.forename: Joo-Won
authors.initials: JW
authors.lastname: Suh
authors.forename: Tae-Ryong
authors.initials: TR
authors.lastname: Hahn
authors.forename: Seong Hee
authors.initials: SH
authors.lastname: Bhoo
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology*; D034301:Deinococcus / Q000276:immunology* / Q000378:metabolism*; D018604:Epitope Mapping / Q000379:methods*; D010834:Phytochrome / Q000737:chemistry* / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D010834:Phytochrome
keywords: DrBphP; epitope; mapping; monoclonal antibody