TY - JOUR
T1 - Determination of critical epitope of PcMab-47 against human podocalyxin
AU - Itai, Shunsuke
AU - Yamada, Shinji
AU - Kaneko, Mika K.
AU - Kato, Yukinari
N1 - Funding Information:
This research was supported in part by AMED under Grant nos.: JP17am0301010 (Y.K.), JP17am0101078 (Y.K.), and JP17ae0101028 (Y.K.), and by JSPS KAKENHI Grant no. 17K07299 (M.K.K.) and Grant no. 16K10748 (Y.K.).
Publisher Copyright:
© 2018 The Authors
Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2018/7
Y1 - 2018/7
N2 - Podocalyxin (PODXL) is a type I transmembrane protein, which is highly glycosylated. PODXL is expressed in some types of human cancer tissues including oral, breast, and lung cancer tissues and may promote tumor growth, invasion, and metastasis. We previously produced PcMab-47, a novel anti-PODXL monoclonal antibody (mAb) which reacts with endogenous PODXL-expressing cancer cell lines and normal cells independently of glycosylation in Western blot, flow cytometry, and immunohistochemical analysis. In this study, we used enzyme-linked immunosorbent assay (ELISA), flow cytometry, and immunohistochemical analysis to determine the epitope of PcMab-47. The minimum epitope of PcMab-47 was found to be Asp207, His208, Leu209, and Met210. A blocking peptide containing this minimum epitope completely neutralized PcMab-47 reaction against oral cancer cells by flow cytometry and immunohistochemical analysis. These findings could lead to the production of more functional anti-PODXL mAbs, which are advantageous for antitumor activities.
AB - Podocalyxin (PODXL) is a type I transmembrane protein, which is highly glycosylated. PODXL is expressed in some types of human cancer tissues including oral, breast, and lung cancer tissues and may promote tumor growth, invasion, and metastasis. We previously produced PcMab-47, a novel anti-PODXL monoclonal antibody (mAb) which reacts with endogenous PODXL-expressing cancer cell lines and normal cells independently of glycosylation in Western blot, flow cytometry, and immunohistochemical analysis. In this study, we used enzyme-linked immunosorbent assay (ELISA), flow cytometry, and immunohistochemical analysis to determine the epitope of PcMab-47. The minimum epitope of PcMab-47 was found to be Asp207, His208, Leu209, and Met210. A blocking peptide containing this minimum epitope completely neutralized PcMab-47 reaction against oral cancer cells by flow cytometry and immunohistochemical analysis. These findings could lead to the production of more functional anti-PODXL mAbs, which are advantageous for antitumor activities.
KW - Epitope mapping
KW - Monoclonal antibody
KW - Oral cancer
KW - PODXL
KW - Podocalyxin
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U2 - 10.1016/j.bbrep.2018.04.003
DO - 10.1016/j.bbrep.2018.04.003
M3 - Article
AN - SCOPUS:85058486663
VL - 14
SP - 78
EP - 82
JO - Biochemistry and Biophysics Reports
JF - Biochemistry and Biophysics Reports
SN - 2405-5808
ER -
