TY - JOUR
T1 - Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses
AU - Kato, Yukinari
AU - Furusawa, Yoshikazu
AU - Yamada, Shinji
AU - Itai, Shunsuke
AU - Takei, Junko
AU - Sano, Masato
AU - Kaneko, Mika K.
N1 - Funding Information:
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 Grant Number 16K10748(Y.K.). We thank Takuro Nakamura, Miyuki Yanaka, Kayo Hisamatsu, Saori Handa, and Yoshimi Nakamura. for their excellent technical assistance.
Publisher Copyright:
© 2019 The Authors
PY - 2019/7
Y1 - 2019/7
N2 - Podoplanin (PDPN) is known as a lymphatic endothelial cell marker. Monoclonal antibodies (mAbs) against human, mouse, rat, rabbit, dog, cat, bovine, pig, and horse PDPN have been established in our previous studies. However, mAbs against alpaca PDPN (aPDPN), required for immunohistochemical analysis, remain to be developed. In the present study, we employed the Cell-Based Immunization and Screening (CBIS) method for producing anti-aPDPN mAbs. We immunized mice with aPDPN-overexpressing Chinese hamster ovary (CHO)-K1 cells (CHO/aPDPN), and hybridomas producing mAbs against aPDPN were screened using flow cytometry. One of the mAbs, PMab-225 (IgG2b, kappa), specifically detected CHO/aPDPN cells via flow cytometry and recognized the aPDPN protein on Western blotting. Further, PMab-225 strongly stained lung type I alveolar cells, colon lymphatic endothelial cells, and kidney podocytes via immunohistochemistry. These findings demonstrate that PMab-225 antibody is useful to investigate the function of aPDPN via different techniques.
AB - Podoplanin (PDPN) is known as a lymphatic endothelial cell marker. Monoclonal antibodies (mAbs) against human, mouse, rat, rabbit, dog, cat, bovine, pig, and horse PDPN have been established in our previous studies. However, mAbs against alpaca PDPN (aPDPN), required for immunohistochemical analysis, remain to be developed. In the present study, we employed the Cell-Based Immunization and Screening (CBIS) method for producing anti-aPDPN mAbs. We immunized mice with aPDPN-overexpressing Chinese hamster ovary (CHO)-K1 cells (CHO/aPDPN), and hybridomas producing mAbs against aPDPN were screened using flow cytometry. One of the mAbs, PMab-225 (IgG2b, kappa), specifically detected CHO/aPDPN cells via flow cytometry and recognized the aPDPN protein on Western blotting. Further, PMab-225 strongly stained lung type I alveolar cells, colon lymphatic endothelial cells, and kidney podocytes via immunohistochemistry. These findings demonstrate that PMab-225 antibody is useful to investigate the function of aPDPN via different techniques.
KW - Alpaca podoplanin
KW - PDPN
KW - PMab-225
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U2 - 10.1016/j.bbrep.2019.100633
DO - 10.1016/j.bbrep.2019.100633
M3 - Article
AN - SCOPUS:85063912933
SN - 2405-5808
VL - 18
JO - Biochemistry and Biophysics Reports
JF - Biochemistry and Biophysics Reports
M1 - 100633
ER -