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Epitope mapping of monoclonal antibodies for the Deinococcus radiodurans bacteriophytochome
Author(s) -
Kim TaeLim,
Yoo Jihey,
Sangsawang Kanidta,
Cho ManHo,
Yang Seung Hwan,
Suh JooWon,
Hahn TaeRyong,
Bhoo Seong Hee
Publication year - 2014
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1002/pro.2464
Subject(s) - epitope , monoclonal antibody , epitope mapping , deinococcus radiodurans , linear epitope , antibody , microbiology and biotechnology , amino acid , recombinant dna , biology , peptide sequence , chemistry , biochemistry , genetics , gene
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.