Production and characterization of monoclonal antibodies directed against connective tissue proteoglycans.
Monoclonal antibodies have been raised against determinants present in cartilage proteoglycan. Characterization of the specificity of these antibodies indicated that they recognize determinants present in the keratan sulfate glycosaminoglycan chain and on chondroitin sulfate oligosaccharide stubs attached to the proteoglycan core protein after chondroitinase digestion of the proteoglycan (i.e., delta-unsaturated 4- and 6-sulfated and unsulfated chondroitin sulfate on the proteoglycan core). The antibody recognizing keratan sulfate has been used to demonstrate the presence of a keratan sulfate-rich proteoglycan subpopulation that increases with increasing age of animal compared with chondroitin sulfate-rich proteoglycans. Monoclonal antibodies recognizing determinants on chondroitinase-treated proteoglycan have been used in immunohistochemical localization studies determining the differential distribution of 4- and 6-sulfated and unsulfated proteoglycans in tissue sections of cartilage and other noncartilaginous tissues. Digestion with chondroitinase ABC or ACII can be used to differentiate between chondroitin sulfate and dermatan sulfate proteoglycan in different connective tissues. In addition, the presence of a 6-sulfated chondroitin sulfate proteoglycan that is associated with membranes surrounding nerve and muscle fiber bundles is described. Monoclonal antibodies were also raised against the link protein(s) of cartilage proteoglycan aggregate. They have been used in peptide map analyses of link protein and in demonstrating the presence of a high-mannose oligosaccharide chain of the link proteins. The presence of high-mannose oligosaccharide structures on the link protein(s) accounts for the microheterogeneity of the link proteins (link proteins 1, 2, or 3) that is observed on sodium dodecyl sulfate-polyacrylamide gels.
Caterson
B
B
Christner
J E
JE
Baker
J R
JR
Couchman
J R
JR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D001665:Binding Sites; D002459:Cell Fusion / Q000379:methods; D003238:Connective Tissue / Q000276:immunology*; D000939:Epitopes; D016326:Extracellular Matrix Proteins; D005455:Fluorescent Antibody Technique; D006651:Histocytochemistry; D006820:Hyaluronic Acid / Q000378:metabolism; D007114:Immunization; D007124:Immunoenzyme Techniques; D007632:Keratan Sulfate / Q000276:immunology; D011134:Polysaccharides / Q000276:immunology; D011485:Protein Binding; D011506:Proteins / Q000276:immunology; D011509:Proteoglycans / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification; D011863:Radioimmunoassay; D051381:Rats; D000099249:Proteoglycan Link Protein
D000911:Antibodies, Monoclonal; D000939:Epitopes; D016326:Extracellular Matrix Proteins; D006820:Hyaluronic Acid; D007632:Keratan Sulfate; D011134:Polysaccharides; D011506:Proteins; D011509:Proteoglycans; D000099249:Proteoglycan Link Protein