Journal
GLYCOBIOLOGY
Volume 25, Issue 9, Pages 953-962Publisher
OXFORD UNIV PRESS INC
DOI: 10.1093/glycob/cwv039
Keywords
binding affinity; chondroitin sulfate E; glycosaminoglycan; monoclonal antibody; surface plasmon resonance
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Funding
- Ministry of Education, Science, Sports, and Culture of Japan [24570168]
- Institute of Tropical Medicine, Nagasaki University
- Grants-in-Aid for Scientific Research [24570168] Funding Source: KAKEN
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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 similar to 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 similar to 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.
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