Epitope Mapping of Monoclonal Antibody PMab-48 Against Dog Podoplanin

Monoclon Antib Immunodiagn Immunother. 2018 Jun;37(3):162-165. doi: 10.1089/mab.2018.0006. Epub 2018 Apr 2.

Abstract

Podoplanin (PDPN), a type I transmembrane sialoglycoprotein, is expressed on normal renal podocytes, pulmonary type I alveolar cells, and lymphatic endothelial cells. Increased expression of PDPN in cancers is associated with poor prognosis and hematogenous metastasis through interactions with C-type lectin-like receptor 2 (CLEC-2) on platelets. We previously reported a novel PMab-48 antibody, which is an anti-dog PDPN (dPDPN) monoclonal antibody (mAb) recognizing PDPN expressed in lymphatic endothelial cells. However, the binding epitope of PMab-48 is yet to be clarified. In this study, an enzyme-linked immunosorbent assay and flow cytometry were used to investigate epitopes of PMab-48. The results revealed that the critical epitope of PMab-48 comprises Asp29, Asp30, Ile31, Ile32, and Pro33 of dPDPN.

Keywords: dPDPN; dog podoplanin; epitope mapping; monoclonal antibody.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Aspartic Acid / chemistry
  • Aspartic Acid / immunology
  • Biomarkers, Tumor / chemistry*
  • Biomarkers, Tumor / genetics
  • Biomarkers, Tumor / immunology
  • CHO Cells
  • Cricetulus
  • Dogs
  • Enzyme-Linked Immunosorbent Assay
  • Epitope Mapping / methods*
  • Epitopes / chemistry*
  • Epitopes / immunology
  • Flow Cytometry
  • Gene Expression
  • Isoleucine / chemistry
  • Isoleucine / immunology
  • Membrane Glycoproteins / chemistry*
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / immunology
  • Point Mutation
  • Proline / chemistry
  • Proline / immunology
  • Protein Binding

Substances

  • Antibodies, Monoclonal
  • Biomarkers, Tumor
  • Epitopes
  • Membrane Glycoproteins
  • Isoleucine
  • Aspartic Acid
  • Proline