Thr80 of Sheep Podoplanin Is a Critical Epitope of the Antisheep Podoplanin Monoclonal Antibody: PMab-256

Monoclon Antib Immunodiagn Immunother. 2020 Jun;39(3):95-100. doi: 10.1089/mab.2020.0010. Epub 2020 May 18.

Abstract

An antisheep podoplanin (sPDPN) monoclonal antibody (mAb), PMab-256, has recently been established. PMab-256 shows positive immunostaining for lymphatic endothelial cells, lung type I alveolar cells, and kidney podocytes. PDPN possesses three platelet-aggregation-stimulating (PLAG) domains, PLAG1, PLAG2, and PLAG3, and a PLAG-like domain (PLD). The binding epitope of many anti-PDPN mAbs is located in PLAG domains or PLD. The purpose of this study is to determine the binding epitope of PMab-256. Analysis of sPDPN deletion mutants revealed that the N-terminus of the PMab-256 epitope exists between amino acids 75 and 80 of sPDPN. Furthermore, analysis of sPDPN point mutations demonstrated that the critical epitope includes Thr80 of sPDPN, indicating that the PMab-256 epitope is in the PLD of sPDPN.

Keywords: PMab-256; epitope; monoclonal antibody; sheep podoplanin.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / immunology*
  • Antibody Specificity / immunology
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Endothelial Cells / immunology
  • Epitope Mapping
  • Epitopes / biosynthesis
  • Epitopes / immunology*
  • Humans
  • Membrane Glycoproteins / immunology*
  • Podocytes / immunology
  • Sheep / immunology*

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • Membrane Glycoproteins
  • PDPN protein, human