TY - JOUR
T1 - Elucidation of critical epitope of anti-rat podoplanin monoclonal antibody PMab-2
AU - Furusawa, Yoshikazu
AU - Yamada, Shinji
AU - Itai, Shunsuke
AU - Nakamura, Takuro
AU - Fukui, Masato
AU - Harada, Hiroyuki
AU - Kaneko, Mika K.
AU - Kato, Yukinari
N1 - Funding Information:
The authors thank Miyuki Yanaka, Saori Handa, Kayo Hisamatsu, and Yoshimi Nakamura for excellent technical assistance. 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 16K10748 (Y.K.).
PY - 2018/8
Y1 - 2018/8
N2 - Rat podoplanin (rPDPN) is a recognized lymphatic endothelial cell marker and is expressed on the podocytes of kidney and type I lung alveolar cells. rPDPN is a type I transmembrane sialoglycoprotein and induces platelet aggregation via the C-type lectin-like receptor-2 of platelets. It comprises four platelet aggregation-stimulating (PLAG) domains: PLAG1-3, present in the N-terminus, and PLAG4, in the center of the PDPN protein. Previously, we developed a mouse anti-rPDPN monoclonal antibody clone, PMab-2, by immunizing the PLAG2 and PLAG3 domains of rPDPN. PMab-2 has applications in Western blot, flow cytometry, and immunohistochemical analyses for detection of both normal and cancer cells. However, the binding epitope of PMab-2 remains to be determined. Herein, we investigated the epitope of PMab-2 using enzyme-linked immunosorbent assay, immunohistochemical analysis, and flow cytometry. The results revealed that the critical epitope of PMab-2 is Leu46 and Glu47 of rPDPN.
AB - Rat podoplanin (rPDPN) is a recognized lymphatic endothelial cell marker and is expressed on the podocytes of kidney and type I lung alveolar cells. rPDPN is a type I transmembrane sialoglycoprotein and induces platelet aggregation via the C-type lectin-like receptor-2 of platelets. It comprises four platelet aggregation-stimulating (PLAG) domains: PLAG1-3, present in the N-terminus, and PLAG4, in the center of the PDPN protein. Previously, we developed a mouse anti-rPDPN monoclonal antibody clone, PMab-2, by immunizing the PLAG2 and PLAG3 domains of rPDPN. PMab-2 has applications in Western blot, flow cytometry, and immunohistochemical analyses for detection of both normal and cancer cells. However, the binding epitope of PMab-2 remains to be determined. Herein, we investigated the epitope of PMab-2 using enzyme-linked immunosorbent assay, immunohistochemical analysis, and flow cytometry. The results revealed that the critical epitope of PMab-2 is Leu46 and Glu47 of rPDPN.
KW - PDPN
KW - PMab-2
KW - epitope mapping
KW - podoplanin
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U2 - 10.1089/mab.2018.0025
DO - 10.1089/mab.2018.0025
M3 - Article
C2 - 30088964
AN - SCOPUS:85052738492
VL - 37
SP - 188
EP - 193
JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
SN - 2167-9436
IS - 4
ER -