A novel approach for preparing disulfide-rich peptide-KLH conjugate applicable to the antibody production

Author:

Katayama Hidekazu1,Mizuno Ryo1,Mita Masatoshi23

Affiliation:

1. Department of Applied Biochemistry, School of Engineering, Tokai University, Kanagawa, Japan

2. Center for Advanced Biomedical Sciences, TWIns, Research Institute for Science and Engineering, Waseda University, Tokyo, Japan

3. Department of Biochemistry, Showa University School of Medicine, Tokyo, Japan

Abstract

ABSTRACT To produce the antiserum against a small peptide, the target peptide-keyhole limpet hemocyanine (KLH) conjugate is generally used as an antigen, although the disulfide-rich peptide-KLH conjugate is still difficult to prepare. In our previous study, we have developed a preparation method of the disulfide-rich peptide-KLH conjugate, and this method was applied to produce the antiserum against a relaxin-like peptide. However, this method is limited to the synthetic peptide antigen, and is not applicable to a native or a recombinant peptide. In this study, to expand the applicability of this method to wide variety of peptides, we newly designed a novel thiol probe enabling the conjugation between various peptides and KLH, and applied it to produce the antiserum against relaxin-like peptide of a starfish Asterias amurensis. The antiserum obtained here showed high antibody-titer and good specificity, strongly suggesting that the method developed in this study is applicable to various peptides.

Funder

Japan Society for the Promotion of Science

Publisher

Oxford University Press (OUP)

Subject

Organic Chemistry,Molecular Biology,Applied Microbiology and Biotechnology,General Medicine,Biochemistry,Analytical Chemistry,Biotechnology

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