TY - JOUR
T1 - Establishment of a Monoclonal Antibody PMab-231 for Tiger Podoplanin
AU - Furusawa, Yoshikazu
AU - Kaneko, Mika K.
AU - Nakamura, Takuro
AU - Itai, Shunsuke
AU - Fukui, Masato
AU - Harada, Hiroyuki
AU - Yamada, Shinji
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:
We thank Miyuki Yanaka, Masato Sano, Kayo Hisamatsu, Saori Handa, and Yoshimi Nakamura for their excellent technical assistance. This research was supported in part by AMED under grant numbers JP18am0101078 (Y.K.), JP18am0301010 (Y.K.), and JP18ae0101028 (Y.K.), and by JSPS KAKENHI grant number 17K07299 (M.K.K.) and 16K10748 (Y.K.).
Publisher Copyright:
© 2019, Mary Ann Liebert, Inc.
PY - 2019/4/1
Y1 - 2019/4/1
N2 - Podoplanin (PDPN), also known as T1alpha, has been used as a lung type I alveolar cell marker in the pathophysiological condition. Although we have established several monoclonal antibodies (mAbs) against mammalian PDPNs, mAbs against tiger PDPN (tigPDPN), which are useful for immunohistochemical analysis, remain to be developed. In this study, we immunized mice with tigPDPN-overexpressing Chinese hamster ovary (CHO)-K1 cells (CHO/tigPDPN) and screened hybridomas producing mAbs against tigPDPN using flow cytometry. One of the mAbs, PMab-231 (IgG2a, kappa), specifically detected CHO/tigPDPN cells using flow cytometry as well as recognized tigPDPN protein using western blotting. In addition, PMab-231 was found to cross-react with cat PDPN (cPDPN). The dissociation constants (KD) of PMab-231 for CHO/tigPDPN and CHO/cPDPN cells were determined to be 1.2 × 10-8 and 1.9 × 10-8, respectively, indicating moderate affinity for CHO/tigPDPN and CHO/cPDPN cells. PMab-231 stained type I alveolar cells of the feline lungs and podocytes of the feline kidneys using immunohistochemistry. Our findings suggest the potential usefulness of PMab-231 for the functional analyses of tigPDPN and cPDPN.
AB - Podoplanin (PDPN), also known as T1alpha, has been used as a lung type I alveolar cell marker in the pathophysiological condition. Although we have established several monoclonal antibodies (mAbs) against mammalian PDPNs, mAbs against tiger PDPN (tigPDPN), which are useful for immunohistochemical analysis, remain to be developed. In this study, we immunized mice with tigPDPN-overexpressing Chinese hamster ovary (CHO)-K1 cells (CHO/tigPDPN) and screened hybridomas producing mAbs against tigPDPN using flow cytometry. One of the mAbs, PMab-231 (IgG2a, kappa), specifically detected CHO/tigPDPN cells using flow cytometry as well as recognized tigPDPN protein using western blotting. In addition, PMab-231 was found to cross-react with cat PDPN (cPDPN). The dissociation constants (KD) of PMab-231 for CHO/tigPDPN and CHO/cPDPN cells were determined to be 1.2 × 10-8 and 1.9 × 10-8, respectively, indicating moderate affinity for CHO/tigPDPN and CHO/cPDPN cells. PMab-231 stained type I alveolar cells of the feline lungs and podocytes of the feline kidneys using immunohistochemistry. Our findings suggest the potential usefulness of PMab-231 for the functional analyses of tigPDPN and cPDPN.
KW - PDPN
KW - PMab-231
KW - tiger podoplanin
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U2 - 10.1089/mab.2019.0003
DO - 10.1089/mab.2019.0003
M3 - Article
C2 - 31009336
AN - SCOPUS:85064664249
VL - 38
SP - 89
EP - 95
JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
SN - 2167-9436
IS - 2
ER -