Isolation and characterization of monoclonal antibodies specific for chondroitin sulfate E

• Published in: Glycobiology (Oxford) 2015-09, Vol.25 (9), p.953-962
• Main Authors: Watanabe, Ippei, Hikita, Tomoya, Mizuno, Haruka, Sekita, Risa, Minami, Akira, Ishii, Ami, Minamisawa, Yuka, Suzuki, Kiyoshi, Maeda, Hiroshi, Hidari, Kazuya I P J, Suzuki, Takashi
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - immunology; Antibody Affinity; Antibody Specificity; Cell Line, Tumor; Chondroitin Sulfates - immunology; Humans; Mice; Rats
• ISSN: 0959-6658; 1460-2423
• DOI: 10.1093/glycob/cwv039

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.