A monoclonal antibody that specifically recognizes a glucuronic acid 2-sulfate-containing determinant in intact chondroitin sulfate chain.
Monoclonal antibodies produced against chick embryo limb bud proteoglycan (PG-M) were selected for their ability to recognize determinants on intact chondroitin sulfate chains. One of these monoclonal antibodies (IgM; designated MO-225) reacts with PG-M, chick embryo cartilage proteoglycans (PG-H, PG-Lb, and PG-Lt), and bovine nasal cartilage proteoglycan, but not with Swarm rat chondrosarcoma proteoglycan. The reactivity of PG-H to MO-225 is not affected by keratanase digestion but is completely abolished after chondroitinase digestion. Competitive binding analyses with various glycosaminoglycan samples indicate that the determinant recognized by MO-225 resides in a D-glucuronic acid 2-sulfate(beta 1----3)N-acetylgalactosamine 6-sulfate disaccharide unit (D-unit) common to antigenic chondroitin sulfates. A tetrasaccharide trisulfate containing D-unit at the reducing end is the smallest chondroitin sulfate fragment that can inhibit the binding of the antibody to PG-H. Decreasing the size of a D-unit-rich chondroitin sulfate by hyaluronidase digestion results in progressive reduction in its inhibitory activity. The results suggest that the epitope has a requirement for a long stretch of a disaccharide-repeating structure for a better fit to the antibody.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002356:Cartilage / Q000032:analysis; D002417:Cattle; D002642:Chick Embryo; D002807:Chondroitin / Q000031:analogs & derivatives*; D002809:Chondroitin Sulfates / Q000276:immunology*; D002810:Chondroitinases and Chondroitin Lyases / Q000378:metabolism; D000939:Epitopes / Q000276:immunology*; D005121:Extremities / Q000032:analysis / Q000196:embryology; D005260:Female; D005965:Glucuronates / Q000276:immunology*; D006026:Glycoside Hydrolases; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D009844:Oligosaccharides / Q000494:pharmacology; D011509:Proteoglycans / Q000276:immunology* / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005965:Glucuronates; D009844:Oligosaccharides; D011509:Proteoglycans; D002807:Chondroitin; D002809:Chondroitin Sulfates; C046100:glucuronic acid 2-sulfate; D006026:Glycoside Hydrolases; C026093:keratan-sulfate endo-1,4-beta-galactosidase; D001616:beta-Galactosidase; D002810:Chondroitinases and Chondroitin Lyases