Novel Monoclonal Antibody LpMab-17 Developed by CasMab Technology Distinguishes Human Podoplanin from Monkey Podoplanin

Yukinari Kato, Satoshi Ogasawara, Hiroharu Oki, Ryusuke Honma, Michiaki Takagi, Yuki Fujii, Takuro Nakamura, Noriko Saidoh, Hazuki Kanno, Mitsuo Umetsu, Satoshi Kamata, Hiroshi Kubo, Mitsuhiro Yamada, Yoshihiko Sawa, Kei Ichi Morita, Hiroyuki Harada, Hiroyoshi Suzuki, Mika Kato Kaneko

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Abstract

Podoplanin (PDPN) is a type-I transmembrane sialoglycoprotein, which possesses a platelet aggregation-stimulating (PLAG) domain in its N-terminus. Among the three PLAG domains, O-glycan on Thr52 of PLAG3 is critical for the binding with C-type lectin-like receptor-2 (CLEC-2) and is essential for platelet-aggregating activity of PDPN. Although many anti-PDPN monoclonal antibodies (mAbs) have been established, almost all mAbs bind to PLAG domains. We recently established CasMab technology to produce mAbs against membranous proteins. Using CasMab technology, we produced a novel anti-PDPN mAb, LpMab-17, which binds to non-PLAG domains. LpMab-17 clearly detected endogenous PDPN of cancer cells and normal cells in Western-blot, flow cytometry, and immunohistochemistry. LpMab-17 recognized glycan-deficient PDPN in flow cytometry, indicating that the interaction between LpMab-17 and PDPN is independent of its glycosylation. The minimum epitope of LpMab-17 was identified as Gly77-Asp82 of PDPN using enzyme-linked immunosorbent assay. Of interest, LpMab-17 did not bind to monkey PDPN, whereas the homology is 94% between human PDPN and monkey PDPN, indicating that the epitope of LpMab-17 is unique compared with the other anti-PDPN mAbs. The combination of different epitope-possessing mAbs could be advantageous for the PDPN-targeting diagnosis or therapy.

Original languageEnglish
Pages (from-to)109-116
Number of pages8
JournalMonoclonal Antibodies in Immunodiagnosis and Immunotherapy
Volume35
Issue number2
DOIs
Publication statusPublished - Apr 1 2016
Externally publishedYes

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ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

Kato, Y., Ogasawara, S., Oki, H., Honma, R., Takagi, M., Fujii, Y., Nakamura, T., Saidoh, N., Kanno, H., Umetsu, M., Kamata, S., Kubo, H., Yamada, M., Sawa, Y., Morita, K. I., Harada, H., Suzuki, H., & Kaneko, M. K. (2016). Novel Monoclonal Antibody LpMab-17 Developed by CasMab Technology Distinguishes Human Podoplanin from Monkey Podoplanin. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, 35(2), 109-116. https://doi.org/10.1089/mab.2015.0077