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Isolation and characterization of monoclonal antibodies specific for chondroitin sulfate E

2015; Oxford University Press; Volume: 25; Issue: 9 Linguagem: Inglês

10.1093/glycob/cwv039

1460-2423

Ippei Watanabe, Tomoya Hikita, Haruka Mizuno, Risa Sekita, Akira Minami, Ami Ishii, Yuka Minamisawa, Kiyoshi Suzuki, Hiroshi Maéda, Kazuya I.P.J. Hidari, Takashi Suzuki,

Skin and Cellular Biology Research

Chondroitin sulfate E (CSE) is a polysaccharide containing mainly disaccharide units of d-glucuronic acid (GlcA) and 4,6-O-disulfated N-acetyl-d-galactosamine (GalNAc) residues (E-unit) in the amount of ∼60%. CSE is involved in many biological and pathological processes. In this study, we established new monoclonal antibodies, termed E-12C and E-18H, by using CSE that contained more than 70% of E-units as an immunogen. These antibodies recognized CSE but not other CSs isomers or dermatan sulfate (DS). We evaluated the reactivities of the antibodies to 6-O-sulfated CSA (6S-CSA) and DS (6S-DS) that possessed ∼60% of GalNAc (4S, 6S) moieties in their structures. Neither of the antibodies reacted with 6S-DS. The antibodies strictly distinguished the structural difference of GlcA and l-iduronic acid in the polysaccharide. Binding affinities of the antibodies were determined by a surface plasmon resonance assay using CSE and 6S-CSA. The binding affinities were strongly associated with the molecular weight of CSE and the E-unit content of 6S-CSA. Moreover, we demonstrated that the antibodies are applicable to histochemical analysis. In conclusion, the new anti-CSE monoclonal antibodies specifically recognize the E-unit of CSE. The antibodies will become useful tools for the investigation of the biological and pathological significance of CSE.