title: Novel Monoclonal Antibody LpMab-17 Developed by CasMab Technology Distinguishes Human Podoplanin from Monkey Podoplanin.
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.
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Yukinari
authors.initials: Y
authors.lastname: Kato
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ogasawara
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .|2 Department of Orthopaedic Surgery, Faculty of Medicine, Yamagata University , Yamagata, Japan .
authors.forename: Hiroharu
authors.initials: H
authors.lastname: Oki
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .|2 Department of Orthopaedic Surgery, Faculty of Medicine, Yamagata University , Yamagata, Japan .
authors.forename: Ryusuke
authors.initials: R
authors.lastname: Honma
authors.affiliation: 2 Department of Orthopaedic Surgery, Faculty of Medicine, Yamagata University , Yamagata, Japan .
authors.forename: Michiaki
authors.initials: M
authors.lastname: Takagi
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Yuki
authors.initials: Y
authors.lastname: Fujii
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Takuro
authors.initials: T
authors.lastname: Nakamura
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Noriko
authors.initials: N
authors.lastname: Saidoh
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .|3 Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University , Sendai, Japan .
authors.forename: Hazuki
authors.initials: H
authors.lastname: Kanno
authors.affiliation: 3 Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University , Sendai, Japan .
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Umetsu
authors.affiliation: 4 Department of Advanced Preventive Medicine for Infectious Disease, Tohoku University Graduate School of Medicine , Sendai, Japan .|5 Department of Thoracic Surgery, Institute of Development, Aging, and Cancer, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kamata
authors.affiliation: 4 Department of Advanced Preventive Medicine for Infectious Disease, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kubo
authors.affiliation: 6 Department of Respiratory Medicine, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Yamada
authors.affiliation: 7 Department of Morphological Biology, Fukuoka Dental College , Fukuoka, Japan .
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Sawa
authors.affiliation: 8 Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University , Tokyo, Japan .
authors.forename: Kei-Ichi
authors.initials: K
authors.lastname: Morita
authors.affiliation: 8 Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University , Tokyo, Japan .
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Harada
authors.affiliation: 9 Department of Pathology and Laboratory Medicine, Sendai Medical Center , Sendai, Japan .
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Suzuki
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Mika Kato
authors.initials: MK
authors.lastname: Kaneko
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D057809:HEK293 Cells; D000882:Haplorhini / Q000276:immunology; D006801:Humans; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D000072417:Protein Domains / Q000276:immunology*; D000098853:Podoplanin
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D008562:Membrane Glycoproteins; C117468:PDPN protein, human; D000098853:Podoplanin