Epitope mapping of anti-mouse podoplanin monoclonal antibody PMab-1
Mouse podoplanin (mPDPN) is a type I transmembrane sialoglycoprotein, which is expressed on lymphatic endothelial cells, podocytes of the kidney, and type I alveolar cells of the lung. mPDPN is known as a platelet aggregation-inducing factor and possesses four platelet aggregation-stimulating (PLAG)...
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doaj-0e87b4bc27a14a1394f0d42adcb0145a2020-11-24T21:33:25ZengElsevierBiochemistry and Biophysics Reports2405-58082018-09-01155256Epitope mapping of anti-mouse podoplanin monoclonal antibody PMab-1Shinji Yamada0Shunsuke Itai1Mika K. Kaneko2Satoru Konnai3Yukinari Kato4Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, JapanDepartment of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, JapanDepartment of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, JapanDepartment of Disease Control, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060-0818, JapanDepartment of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, Japan; New Industry Creation Hatchery Center, Tohoku University, 2-1, Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, Japan; Corresponding author at: New Industry Creation Hatchery Center, Tohoku University, 2-1, Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, Japan.Mouse podoplanin (mPDPN) is a type I transmembrane sialoglycoprotein, which is expressed on lymphatic endothelial cells, podocytes of the kidney, and type I alveolar cells of the lung. mPDPN is known as a platelet aggregation-inducing factor and possesses four platelet aggregation-stimulating (PLAG) domains: PLAG1, PLAG2, and PLAG3 in the N-terminus and PLAG4 in the middle of the mPDPN protein. mPDPN overexpression in cancers has been reportedly associated with hematogenous metastasis through interaction with the C-type lectin-like receptor 2 of platelets. We previously reported a rat anti-mPDPN monoclonal antibody clone PMab-1, which was developed by immunizing the PLAG2 and PLAG3 domains of mPDPN. PMab-1 is very useful in flow cytometry, western blot, and immunohistochemical analyses to detect both normal cells and cancers. However, the binding epitope of PMab-1 remains to be clarified. In the present study, flow cytometry, enzyme-linked immunosorbent assay, and immunohistochemical analyses were utilized to investigate the epitope of PMab-1. The results revealed that the critical epitope of PMab-1 is Asp39 and Met41 of mPDPN. These findings can be applied to the production of more functional anti-mPDPN monoclonal antibodies. Keywords: Podoplanin, PDPN, PMab-1, Epitope mappinghttp://www.sciencedirect.com/science/article/pii/S2405580818301493 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Shinji Yamada Shunsuke Itai Mika K. Kaneko Satoru Konnai Yukinari Kato |
spellingShingle |
Shinji Yamada Shunsuke Itai Mika K. Kaneko Satoru Konnai Yukinari Kato Epitope mapping of anti-mouse podoplanin monoclonal antibody PMab-1 Biochemistry and Biophysics Reports |
author_facet |
Shinji Yamada Shunsuke Itai Mika K. Kaneko Satoru Konnai Yukinari Kato |
author_sort |
Shinji Yamada |
title |
Epitope mapping of anti-mouse podoplanin monoclonal antibody PMab-1 |
title_short |
Epitope mapping of anti-mouse podoplanin monoclonal antibody PMab-1 |
title_full |
Epitope mapping of anti-mouse podoplanin monoclonal antibody PMab-1 |
title_fullStr |
Epitope mapping of anti-mouse podoplanin monoclonal antibody PMab-1 |
title_full_unstemmed |
Epitope mapping of anti-mouse podoplanin monoclonal antibody PMab-1 |
title_sort |
epitope mapping of anti-mouse podoplanin monoclonal antibody pmab-1 |
publisher |
Elsevier |
series |
Biochemistry and Biophysics Reports |
issn |
2405-5808 |
publishDate |
2018-09-01 |
description |
Mouse podoplanin (mPDPN) is a type I transmembrane sialoglycoprotein, which is expressed on lymphatic endothelial cells, podocytes of the kidney, and type I alveolar cells of the lung. mPDPN is known as a platelet aggregation-inducing factor and possesses four platelet aggregation-stimulating (PLAG) domains: PLAG1, PLAG2, and PLAG3 in the N-terminus and PLAG4 in the middle of the mPDPN protein. mPDPN overexpression in cancers has been reportedly associated with hematogenous metastasis through interaction with the C-type lectin-like receptor 2 of platelets. We previously reported a rat anti-mPDPN monoclonal antibody clone PMab-1, which was developed by immunizing the PLAG2 and PLAG3 domains of mPDPN. PMab-1 is very useful in flow cytometry, western blot, and immunohistochemical analyses to detect both normal cells and cancers. However, the binding epitope of PMab-1 remains to be clarified. In the present study, flow cytometry, enzyme-linked immunosorbent assay, and immunohistochemical analyses were utilized to investigate the epitope of PMab-1. The results revealed that the critical epitope of PMab-1 is Asp39 and Met41 of mPDPN. These findings can be applied to the production of more functional anti-mPDPN monoclonal antibodies. Keywords: Podoplanin, PDPN, PMab-1, Epitope mapping |
url |
http://www.sciencedirect.com/science/article/pii/S2405580818301493 |
work_keys_str_mv |
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