Characterization of the Anti-Bovine Podoplanin Monoclonal Antibody PMab-44

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Shinji YamadaYukinari Kato

Abstract

A type I transmembrane sialoglycoprotein podoplanin (PDPN) is expressed in several normal cells, including podocytes of the kidney, type I alveolar cells of the lung, and lymphatic endothelial cells. We recently produced an anti-bovine PDPN (bovPDPN) monoclonal antibody (mAb), PMab-44, by immunizing mice with recombinant proteins of bovPDPN. In this study, we determined the critical epitope of PMab-44 for the recognition of bovPDPN using many deletion mutants and point mutants of bovPDPN. Flow cytometric analyses revealed that the epitope of PMab-44 was Glu46-Thr50, which corresponds to platelet aggregation-stimulating (PLAG) domain-3. The important amino acids in the PMab-44 epitope were determined to be Glu46, Tyr48, and Thr50. Western blot analysis also confirmed these results, indicating that the PLAG domain of bovPDPN is also important in immunogenicity for producing useful anti-PDPN mAbs.

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Citations

Dec 21, 2018·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Yukinari KatoMika K Kaneko
Dec 21, 2018·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Yukinari KatoMika K Kaneko
Nov 21, 2018·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Shinji YamadaYukinari Kato
Oct 16, 2019·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Yusuke SayamaYukinari Kato
Oct 31, 2019·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Mika K KanekoYukinari Kato
Jan 15, 2020·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Yusuke SayamaYukinari Kato
Apr 3, 2020·Monoclonal Antibodies in Immunodiagnosis and Immunotherapy·Yusuke SayamaYukinari Kato

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Methods Mentioned

BETA
flow cytometry

Software Mentioned

GraphPad PRISM

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