Characterization of monoclonal antibody LpMab-3 recognizing sialylated glycopeptide of podoplanin.
Podoplanin (PDPN/Aggrus/T1α/gp36/OTS-8), a type I transmembrane sialoglycoprotein, is involved in platelet aggregation, cell invasion, and cancer metastasis. Podoplanin expression in cancer cells or cancer-associated fibroblasts was reported to be involved in poor prognosis of several cancers. Furthermore, podoplanin is expressed in lymphatic endothelial cells or lung type I alveolar cells. Although many anti-podoplanin monoclonal antibodies (MAbs), such as NZ-1 and D2-40, have been established, almost all anti-podoplanin MAbs are produced against a platelet aggregation-inducing (PLAG) domain. In this study, we produced and characterized a novel anti-podoplanin monoclonal antibody, LpMab-3, the epitope of which is a sialylated glycopeptide of podoplanin. We identified the minimum epitope of LpMab-3 as Thr76-Glu81 of human podoplanin, which is different from PLAG domain, using Western blot analysis and flow cytometry. Immunohistochemical analysis showed that LpMab-3 is useful for detecting lung type I alveolar cells and lymphatic endothelial cells. Because LpMab-3 detects only sialylated podoplanin, it could be useful for uncovering the physiological function of sialylated human podoplanin.
1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Miyagi, Japan .
Hiroharu
H
Oki
Satoshi
S
Ogasawara
Mika Kato
MK
Kaneko
Michiaki
M
Takagi
Masanori
M
Yamauchi
Yukinari
Y
Kato
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry; D000918:Antibody Specificity; D015153:Blotting, Western; D016466:CHO Cells; D002240:Carbohydrate Sequence; D045744:Cell Line, Tumor; D003412:Cricetulus; D042783:Endothelial Cells / Q000276:immunology / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D004847:Epithelial Cells / Q000276:immunology / Q000473:pathology; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007114:Immunization; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011650:Pulmonary Alveoli / Q000276:immunology / Q000473:pathology; D012795:Sialoglycoproteins / Q000032:analysis* / Q000276:immunology; D000098853:Podoplanin
D000911:Antibodies, Monoclonal; D008562:Membrane Glycoproteins; D012795:Sialoglycoproteins; C117468:PDPN protein, human; D000098853:Podoplanin