Elucidation of critical epitope of anti-rat podoplanin monoclonal antibody PMab-2

Yoshikazu Furusawa, Shinji Yamada, Shunsuke Itai, Takuro Nakamura, Masato Fukui, Hiroyuki Harada, Mika K. Kaneko, Yukinari Kato

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Rat podoplanin (rPDPN) is a recognized lymphatic endothelial cell marker and is expressed on the podocytes of kidney and type I lung alveolar cells. rPDPN is a type I transmembrane sialoglycoprotein and induces platelet aggregation via the C-type lectin-like receptor-2 of platelets. It comprises four platelet aggregation-stimulating (PLAG) domains: PLAG1-3, present in the N-terminus, and PLAG4, in the center of the PDPN protein. Previously, we developed a mouse anti-rPDPN monoclonal antibody clone, PMab-2, by immunizing the PLAG2 and PLAG3 domains of rPDPN. PMab-2 has applications in Western blot, flow cytometry, and immunohistochemical analyses for detection of both normal and cancer cells. However, the binding epitope of PMab-2 remains to be determined. Herein, we investigated the epitope of PMab-2 using enzyme-linked immunosorbent assay, immunohistochemical analysis, and flow cytometry. The results revealed that the critical epitope of PMab-2 is Leu46 and Glu47 of rPDPN.

Original languageEnglish
Pages (from-to)188-193
Number of pages6
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume37
Issue number4
DOIs
Publication statusPublished - 2018 Aug

Keywords

  • PDPN
  • PMab-2
  • epitope mapping
  • podoplanin

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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