Epitope Mapping of PMab-241, a Lymphatic Endothelial Cell-Specific Anti-Bear Podoplanin Monoclonal Antibody

Yusuke Sayama, Masato Sano, Teizo Asano, Yoshikazu Furusawa, Junko Takei, Takuro Nakamura, Miyuki Yanaka, Saki Okamoto, Saori Handa, Yu Komatsu, Yoshimi Nakamura, Mikiko Yanagawa, Mika K. Kaneko, Yukinari Kato

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Abstract

Anti-bear podoplanin (bPDPN) monoclonal antibodies (mAbs), including PMab-247 and PMab-241, have been previously established. Although PMab-247 has shown positive immunostaining for lymphatic endothelial cells (LECs), type I alveolar cells of the lung, and podocytes of the kidney, PMab-241 stains LECs but does not react with lung type I alveolar cells. PDPN possesses three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) and the PLAG-like domain (PLD). The binding epitope of PMab-247 was previously determined to include bPDPN residues Asp76, Arg78, Glu80, and Arg82. Among these, Glu80 and Arg82 are included in PLD of bPDPN. The purpose of this study is to determine the binding epitope of PMab-241 and to clarify the difference between these two anti-bPDPN mAbs. Analysis of bPDPN deletion mutants revealed that the N-terminus of the PMab-241 epitope exists between amino acids (aa) 75 and 80 of bPDPN. In addition, analysis of bPDPN point mutants demonstrated that the critical epitope of PMab-241 includes Thr75, Asp76, and Arg78 of bPDPN. The binding epitopes of PMab-241 and PMab-247 seem to overlap, but this slight difference may be sufficient to provide the specificity of PMab-241 to discriminate LECs from type I alveolar cells of the lung.

Original languageEnglish
Pages (from-to)77-81
Number of pages5
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume39
Issue number3
DOIs
Publication statusPublished - 2020 Jun

Keywords

  • PMab-241
  • bear podoplanin
  • epitope
  • monoclonal antibody

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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    Sayama, Y., Sano, M., Asano, T., Furusawa, Y., Takei, J., Nakamura, T., Yanaka, M., Okamoto, S., Handa, S., Komatsu, Y., Nakamura, Y., Yanagawa, M., Kaneko, M. K., & Kato, Y. (2020). Epitope Mapping of PMab-241, a Lymphatic Endothelial Cell-Specific Anti-Bear Podoplanin Monoclonal Antibody. Monoclonal antibodies in immunodiagnosis and immunotherapy, 39(3), 77-81. https://doi.org/10.1089/mab.2020.0004