A sulfanyl-PEG derivative of relaxin-like peptide utilizable for the conjugation with KLH and the antibody production.
A small peptide-keyhole limpet hemocyanin (KLH) conjugate is generally used as an antigen for producing specific antibodies. However, preparation of a disulfide-rich heterodimeric peptide-KLH conjugates is difficult. In this study, we developed a novel method for preparation of the conjugate, and applied it to the production of specific antibodies against the relaxin-like gonad-stimulating peptide (RGP) from the starfish. In this method, a sulfanyl group necessary for the conjugation with KLH was site-specifically introduced to the peptide after regioselective disulfide bond formation reactions. Using the conjugate, we could obtain specific antibodies with a high antibody titer. This method might also be useful for the production of antibodies against other heterodimeric peptides with disulfide cross-linkages, such as vertebrate relaxins.
Department of Applied Biochemistry, School of Engineering, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan. Electronic address: katay@tokai-u.jp.
Hidekazu
H
Katayama
Department of Biology, Faculty of Education, Tokyo Gakugei University, 4-1-1 Nukuikita-machi, Koganei, Tokyo 184-8501, Japan.
Masatoshi
M
Mita
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation / Q000187:drug effects*; D006433:Hemocyanins / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D066244:Proton Magnetic Resonance Spectroscopy; D012065:Relaxin / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013215:Starfish
D011092:Polyethylene Glycols; D012065:Relaxin; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin
Antigen; PEGylation; Relaxin-like gonad-stimulating peptide; Solid-phase peptide synthesis; Starfish