Epitope Mapping of Anti-Bear Podoplanin Monoclonal Antibody PMab-247

Yukinari Kato, Junko Takei, Yoshikazu Furusawa, Yusuke Sayama, Masato Sano, Satoru Konnai, Atsushi Kobayashi, Hiroyuki Harada, Maki Takahashi, Hiroyoshi Suzuki, Shinji Yamada, Mika K. Kaneko

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)

Abstract

Podoplanin (PDPN)/T1alpha is a type I transmembrane sialoglycoprotein, which is expressed on podocytes of the kidneys and type I alveolar cells of the lungs. PDPN is also known as Aggrus, a platelet aggregation-inducing factor, which comprises three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) in the N-terminus and PLAG-like domains (PLDs) in the middle of the PDPN protein. We have previously established a mouse anti-bear PDPN (bPDPN) monoclonal antibody (mAb) clone, PMab-247 using the Cell-Based Immunization and Screening (CBIS) method. PMab-247 is very useful in flow cytometry, Western blotting, and immunohistochemical (IHC) analyses; however, the binding epitope of PMab-247 has not been elucidated. In this study, we aimed to investigate the epitope of PMab-247 using enzyme-linked immunosorbent assay and IHC analyses. The results revealed that the critical epitopes of PMab-247 are Asp76, Arg78, Glu80, and Arg82 of bPDPN. The Glu80 and Arg82 are included in PLD of bPDPN. The findings of our study can be applied to the production of more functional anti-bPDPN mAbs.

Original languageEnglish
Pages (from-to)230-233
Number of pages4
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume38
Issue number5
DOIs
Publication statusPublished - 2019 Oct

Keywords

  • PDPN
  • PMab-247
  • epitope mapping
  • podoplanin

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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