Preparation and identification of a population of antibodies that recognize carbodiimide-modified heparin.
Protein-heparin complexes, prepared by a water-soluble carbodiimide coupling technique, were used to produce anti-heparin antibodies in rabbits. Antiserums that recognized carbodiimide-treated heparin, but not untreated heparin, were obtained. Carbodiimide-treated heparan sulfate exhibited 10% to 20% cross-reactivity compared with a similarly treated heparin, whereas there was no cross-reactivity with five other carbodiimide-treated mucopolysaccharides. 3H-1-ethyl-3-(3-trimethylammoniumpropyl) carbodiimide iodide was used to demonstrate that carbodiimide forms a stable adduct with heparin and other mucopolysaccharides. Using an antibody fraction that eluted from 1-ethyl-3-(3-trimethylammoniumpropyl) carbodiimide iodide-treated heparin-Sepharose with 2 mol/L KI, it was demonstrated that, for the antibody population studied, the addition of one carbodiimide per heparin molecule resulted in complete epitope expression without loss of anticoagulant activity. The addition of up to eight additional carbodiimide molecules to heparin did not increase the extent of epitope formation, although anticoagulant activity was lost. Except for heparan sulfate, the addition of radiolabeled carbodiimide to other mucopolysaccharides did not result in epitope formation. These data demonstrate that antibodies to an epitope derived from heparin can be formed, that the epitope is fully expressed while anticoagulant activity is present, and that the antibody is specifically directed against an altered portion of the polysaccharide.
Gitel
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Medina
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VM
Wessler
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S
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D002234:Carbodiimides / Q000494:pharmacology*; D002809:Chondroitin Sulfates; D003429:Cross Reactions; D003871:Dermatan Sulfate; D006025:Glycosaminoglycans / Q000276:immunology; D006385:Hemagglutination Inhibition Tests; D006493:Heparin / Q000276:immunology*; D006497:Heparan Sulfate; D006820:Hyaluronic Acid; D007074:Immunoglobulin G / Q000302:isolation & purification; D007632:Keratan Sulfate; D011817:Rabbits; D013997:Time Factors
D000906:Antibodies; D002234:Carbodiimides; D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D006025:Glycosaminoglycans; D006493:Heparin; D006497:Heparan Sulfate; D006820:Hyaluronic Acid; D007074:Immunoglobulin G; D007632:Keratan Sulfate