Monoclonal antibodies reacting with endo-beta-galactosidase-resistant epitopes on keratan sulfate-bearing fragments of bovine nasal cartilage proteoglycan.
Six monoclonal antibody-producing cell lines were derived from mice immunized with keratan sulfate (KS)-bearing tryptic fragments of the core protein of bovine nasal cartilage proteoglycan monomer digested with KS-specific endo-beta-galactosidase. The monoclonal antibodies were characterized by solid-phase ELISA competition studies and SDS-PAGE immunoblotting. Two of them resemble previously described monoclonal antibodies that are directed to epitopes containing both KS and core protein. In contrast, the remaining four monoclonal antibodies are unprecedented in being directed to epitopes whose reactivity is unaffected or enhanced by endo-beta-galactosidase degradation of KS. SDS-PAGE immunoblots revealed two large KS-bearing tryptic fragments of cartilage proteoglycan and a heterogeneous population of smaller fragments not evident by non-immunological techniques. Some of the antibodies used react with all KS-bearing fragments, others react only with the two largest fragments.
Department of Medicine, Albert Einstein College of Medicine, Bronx, New York 10461.
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HD
Keiser
D053674:Aggrecans; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001667:Binding, Competitive; D002356:Cartilage / Q000737:chemistry*; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology* / Q000378:metabolism; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins / Q000276:immunology* / Q000378:metabolism; D006026:Glycoside Hydrolases; D015151:Immunoblotting; D007632:Keratan Sulfate / Q000276:immunology* / Q000378:metabolism; D037181:Lectins, C-Type; D009300:Nasal Septum / Q000737:chemistry; D010446:Peptide Fragments / Q000276:immunology* / Q000378:metabolism; D011509:Proteoglycans; D014357:Trypsin; D001616:beta-Galactosidase / Q000378:metabolism*
D053674:Aggrecans; D000911:Antibodies, Monoclonal; D000939:Epitopes; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D037181:Lectins, C-Type; D010446:Peptide Fragments; D011509:Proteoglycans; D007632:Keratan Sulfate; D006026:Glycoside Hydrolases; C026093:keratan-sulfate endo-1,4-beta-galactosidase; D001616:beta-Galactosidase; D014357:Trypsin