TY - JOUR
T1 - Characterization of the Anti-Bovine Podoplanin Monoclonal Antibody PMab-44
AU - Yamada, Shinji
AU - Honma, Ryusuke
AU - Kaneko, Mika K.
AU - Nakamura, Takuro
AU - Yanaka, Miyuki
AU - Saidoh, Noriko
AU - Takagi, Michiaki
AU - Konnai, Satoru
AU - Kato, Yukinari
N1 - Funding Information:
This research was supported in part by grants from the Science and Technology Research Promotion Program for Agriculture, Forestry, Fisheries, and Food Industry, Japan (grant 26058B to S.K.), and the NARO, Bio-oriented Technology Research Advancement Institution (the special scheme project on regional developing strategy: grant 16817557 to S.K.). This work was also supported in part by Project for utilizing glycans in the development of innovative drug discovery technologies from Japan Agency for Medical Research and Development, AMED (Y.K.), by the Basic Science and Platform Technology Program for Innovative Biological Medicine from AMED (Y.K.), by the Platform for Drug Discovery, Informatics, and Structural Life Science (PDIS) from AMED (Y.K.), by the Regional Innovation Strategy Support Program from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan (Y.K.), by JSPS KAKENHI Grant Number 26440019 and 17K07299 (M.K.K.) and 16K10748 (Y.K.). This work was performed in part under the Cooperative Research Program of Institute for Protein Research, Osaka University, CR-16-05 and CR-17-05 and by the Grant for Joint Research Project of the Institute of Medical Science, the University of Tokyo.
Publisher Copyright:
© Copyright 2017, Mary Ann Liebert, Inc. 2017.
PY - 2017/6
Y1 - 2017/6
N2 - 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.
AB - 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.
KW - Bovine PDPN
KW - PMab-44
KW - Podoplanin
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U2 - 10.1089/mab.2017.0016
DO - 10.1089/mab.2017.0016
M3 - Article
C2 - 28498096
AN - SCOPUS:85021115336
SN - 2167-9436
VL - 36
SP - 129
EP - 134
JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
IS - 3
ER -