Identification of a target antigen recognized by a mouse monoclonal antibody to the bile canalicular surface of rat hepatocytes with a random phage display library

Masanori Ariyoshi, Motowo Mizuno, Yoshiko Morisue, Morizou Shimada, Shirou Fujita, Junichirou Nasu, Hiroyuki Okada, Hiroyuki Shimomura, Kazuhide Yamamoto, Takao Tsuji

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We developed a monoclonal antibody (MoAb) (clone 5E8) against an antigen on the bile canalicular membrane of rat hepatocyte. By immunoblotting, MoAb 5E8 detected a band of 110 kD. In this study, we used the phage display technique to identify the target antigen recognized by MoAb 5E8. We screened a random phage display library expressing 12-mer peptide sequences and identified a peptide sequence, FHFNPYTGHPLT, as an epitope. We compared this peptide sequence with those of dipeptidyl peptidase IV (DPP IV, E.C.3.4.14.5) and Cell-CAM105, which proteins were located by a database search based on the information of tissue localization and approximate molecular weight of the MoAb 5E8 antigen, and sequence similarity with a region in DPP IV (amino acids 225 233) but not with Cell-CAM105 was found. In addition, we immunohistochemically stained various tissues (liver, small intestine, and kidney) of Japanese Fischer 344 rats, known to be deficient for DPP IV, with MoAb 5E8 and showed that the expression of MoAb 5E8 antigen was negligible or weak. In contrast, tissues sampled from the same organs of Sprague-Dawley rats, known to express DPP IV, were positively stained. These findings suggest that the antigen recognized by MoAb 5E8 is DDPIV and its major epitope is located in amino acids at positions 225-233.

Original languageEnglish
Pages (from-to)187-191
Number of pages5
JournalActa medica Okayama
Volume56
Issue number4
Publication statusPublished - Dec 1 2002

Keywords

  • Bile canalicular membrane
  • Dipeptidyl peptidase IV
  • Epitope
  • Monoclonal antibody
  • Random phage display library

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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    Ariyoshi, M., Mizuno, M., Morisue, Y., Shimada, M., Fujita, S., Nasu, J., Okada, H., Shimomura, H., Yamamoto, K., & Tsuji, T. (2002). Identification of a target antigen recognized by a mouse monoclonal antibody to the bile canalicular surface of rat hepatocytes with a random phage display library. Acta medica Okayama, 56(4), 187-191.