Establishment of Mouse Monoclonal Antibody LpMab-13 Against Human Podoplanin

Satoshi Ogasawara, Mika K. Kaneko, Ryusuke Honma, Hiroharu Oki, Yuki Fujii, Michiaki Takagi, Hiroyoshi Suzuki, Yukinari Kato

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

Podoplanin (PDPN)/Aggrus is a type-I transmembrane sialoglycoprotein, which possesses a platelet aggregation-stimulating (PLAG) domain. The O-glycosylation on Thr52 of human PDPN (hPDPN) is critical for the interaction of hPDPN with C-type lectin-like receptor-2 (CLEC-2), resulting in platelet aggregation. Many anti-hPDPN monoclonal antibodies (MAbs) against PLAG domains and non-PLAG domains have been established; however, mouse anti-PLAG2/3 MAb, the epitope of which is consistent with rat anti-PLAG2/3 MAb NZ-1, has not been established. NZ-1 inhibits the hPDPN-CLEC-2 interaction and is also useful for anti-PA tag MAb. We recently established CasMab technology to produce MAbs against membranous proteins. Herein, we produced a novel anti-hPDPN MAb, LpMab-13, which binds to PLAG2/3 domains. LpMab-13 recognized endogenous hPDPN of cancer cells, including glioblastoma, oral cancer, lung cancer, and malignant mesothelioma, and normal cells such as lymphatic endothelial cells and podocytes of kidney in Western blot, flow cytometry, and immunohistochemistry. LpMab-13 recognized glycan-deficient hPDPN in flow cytometry, indicating that the interaction between LpMab-13 and hPDPN is independent of its glycosylation. The minimum epitope of LpMab-13 was identified as Ala42-Asp49 of hPDPN using Western blot and flow cytometry. The combination of different epitope-possessing MAbs could be advantageous for the hPDPN-targeting diagnosis and therapy.

Original languageEnglish
Pages (from-to)155-162
Number of pages8
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume35
Issue number3
DOIs
Publication statusPublished - 2016 Jun 1

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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