Thrombolysis by a fibrin-specific antibody Fab'-urokinase conjugate.
The unidirectional crosslinking agent N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP) was used to covalently couple fibrin-specific antibody Fab' to low-molecular-weight urokinase by means of the sulfhydryl groups of the inter-heavy-chain disulfide bonds. At enzymatic activities equal with respect to the substrate pyroglutamyl-glycyl-arginyl-p-nitroanilide-hydrochloride, the Fab'-urokinase conjugate lysed fibrin monomer linked to sepharose with a potency 95 times that of urokinase (which is essentially identical to the potency previously reported for an intact antibody-urokinase conjugate). The Fab'-urokinase conjugate also lysed human thrombi in fresh citrated plasma with a maximal potency 4.4 times that of uncoupled urokinase. These findings indicate that univalent antibody binding is sufficient to enhance fibrinolysis and that the increased potency of antifibrin-urokinase conjugates can also be demonstrated when cross-linked thrombi are used as substrate in the presence of plasma.
D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D004796:Clinical Enzyme Tests; D003432:Cross-Linking Reagents / Q000494:pharmacology; D005337:Fibrin / Q000276:immunology* / Q000378:metabolism; D005342:Fibrinolysis; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D013379:Substrate Specificity; D013388:Succinimides / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000502:physiology*
D000906:Antibodies; D003432:Cross-Linking Reagents; D007140:Immunoglobulin Fab Fragments; D013388:Succinimides; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate; D005337:Fibrin; D014568:Urokinase-Type Plasminogen Activator