TY - JOUR
T1 - Characterization of Anti-Goat Podoplanin Monoclonal Antibody PMab-235 Using Immunohistochemistry Against Goat Tissues
AU - Takei, Junko
AU - Itai, Shunsuke
AU - Harada, Hiroyuki
AU - Furusawa, Yoshikazu
AU - Miwa, Takashi
AU - Fukui, Masato
AU - Nakamura, Takuro
AU - Sano, Masato
AU - Sayama, Yusuke
AU - Yanaka, Miyuki
AU - Handa, Saori
AU - Hisamatsu, Kayo
AU - Nakamura, Yoshimi
AU - Yamada, Shinji
AU - Kaneko, Mika K.
AU - Kato, Yukinari
N1 - Funding Information:
Y.K. received research funding from ZENOAQ RESOURCE CO., LTD. The other authors have no conflict of interest.
Funding Information:
This research was supported in part by AMED under Grant Nos. JP18am0101078 (Y.K.), JP18am0301010 (Y.K.), and JP18ae0101028 (Y.K.), and by JSPS KAKENHI Grant Nos. 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 expressed on lymphatic endothelial cells, type I alveolar cells of the lungs, and podocytes of the kidney. PDPN possesses three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) of the N-terminus and the PLAG-like domains (PLDs). We previously reported an anti-goat PDPN (gPDPN) monoclonal antibody (mAb), PMab-235, which was developed using the Cell-Based Immunization and Screening (CBIS) method. PMab-235 is very useful in flow cytometry, Western blotting, and immunohistochemical analyses; however, the binding epitope of PMab-235 remains to be elucidated. In this study, we investigated the epitopes of PMab-235 using enzyme-linked immunosorbent assay and immunohistochemistry. The results revealed that the critical epitope of PMab-235 produced by CBIS method is Arg75, Leu78, and Pro79 of gPDPN, which is included in PLD. The findings of our study can be applied to the production of more functional anti-gPDPN mAbs.
AB - Podoplanin (PDPN)/T1alpha is expressed on lymphatic endothelial cells, type I alveolar cells of the lungs, and podocytes of the kidney. PDPN possesses three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) of the N-terminus and the PLAG-like domains (PLDs). We previously reported an anti-goat PDPN (gPDPN) monoclonal antibody (mAb), PMab-235, which was developed using the Cell-Based Immunization and Screening (CBIS) method. PMab-235 is very useful in flow cytometry, Western blotting, and immunohistochemical analyses; however, the binding epitope of PMab-235 remains to be elucidated. In this study, we investigated the epitopes of PMab-235 using enzyme-linked immunosorbent assay and immunohistochemistry. The results revealed that the critical epitope of PMab-235 produced by CBIS method is Arg75, Leu78, and Pro79 of gPDPN, which is included in PLD. The findings of our study can be applied to the production of more functional anti-gPDPN mAbs.
KW - epitope mapping
KW - PDPN
KW - PMab-235
KW - podoplanin
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U2 - 10.1089/mab.2019.0022
DO - 10.1089/mab.2019.0022
M3 - Article
C2 - 31403389
AN - SCOPUS:85073182528
SN - 2167-9436
VL - 38
SP - 213
EP - 219
JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
IS - 5
ER -