TY - JOUR
T1 - Epitope Mapping of Anti-Bear Podoplanin Monoclonal Antibody PMab-247
AU - Kato, Yukinari
AU - Takei, Junko
AU - Furusawa, Yoshikazu
AU - Sayama, Yusuke
AU - Sano, Masato
AU - Konnai, Satoru
AU - Kobayashi, Atsushi
AU - Harada, Hiroyuki
AU - Takahashi, Maki
AU - Suzuki, Hiroyoshi
AU - Yamada, Shinji
AU - Kaneko, Mika K.
N1 - Funding Information:
This research was supported in part by AMED under grant numbers JP19am0101078 (Y.K.), JP18am0301010 (Y.K.), and JP19ae0101028 (Y.K.), and by JSPS KA-KENHI grant numbers 17K07299 (M.K.K.) and 19K07705 (Y.K.).
Publisher Copyright:
© 2019 Mary Ann Liebert, Inc., publishers.
PY - 2019/10
Y1 - 2019/10
N2 - Podoplanin (PDPN)/T1alpha is a type I transmembrane sialoglycoprotein, which is expressed on podocytes of the kidneys and type I alveolar cells of the lungs. PDPN is also known as Aggrus, a platelet aggregation-inducing factor, which comprises three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) in the N-terminus and PLAG-like domains (PLDs) in the middle of the PDPN protein. We have previously established a mouse anti-bear PDPN (bPDPN) monoclonal antibody (mAb) clone, PMab-247 using the Cell-Based Immunization and Screening (CBIS) method. PMab-247 is very useful in flow cytometry, Western blotting, and immunohistochemical (IHC) analyses; however, the binding epitope of PMab-247 has not been elucidated. In this study, we aimed to investigate the epitope of PMab-247 using enzyme-linked immunosorbent assay and IHC analyses. The results revealed that the critical epitopes of PMab-247 are Asp76, Arg78, Glu80, and Arg82 of bPDPN. The Glu80 and Arg82 are included in PLD of bPDPN. The findings of our study can be applied to the production of more functional anti-bPDPN mAbs.
AB - Podoplanin (PDPN)/T1alpha is a type I transmembrane sialoglycoprotein, which is expressed on podocytes of the kidneys and type I alveolar cells of the lungs. PDPN is also known as Aggrus, a platelet aggregation-inducing factor, which comprises three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) in the N-terminus and PLAG-like domains (PLDs) in the middle of the PDPN protein. We have previously established a mouse anti-bear PDPN (bPDPN) monoclonal antibody (mAb) clone, PMab-247 using the Cell-Based Immunization and Screening (CBIS) method. PMab-247 is very useful in flow cytometry, Western blotting, and immunohistochemical (IHC) analyses; however, the binding epitope of PMab-247 has not been elucidated. In this study, we aimed to investigate the epitope of PMab-247 using enzyme-linked immunosorbent assay and IHC analyses. The results revealed that the critical epitopes of PMab-247 are Asp76, Arg78, Glu80, and Arg82 of bPDPN. The Glu80 and Arg82 are included in PLD of bPDPN. The findings of our study can be applied to the production of more functional anti-bPDPN mAbs.
KW - epitope mapping
KW - PDPN
KW - PMab-247
KW - podoplanin
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U2 - 10.1089/mab.2019.0025
DO - 10.1089/mab.2019.0025
M3 - Article
C2 - 31535919
AN - SCOPUS:85073183155
SN - 2167-9436
VL - 38
SP - 230
EP - 233
JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
IS - 5
ER -