title: Antibody-directed fibrinolysis. An antibody specific for both fibrin and tissue plasminogen activator.
abstract: An investigation was made to determine whether it is possible to attract tissue plasminogen activator (tPA) to the site of a thrombus by means of an antibody with affinites for both tPA and fibrin. A bispecific antibody conjugate was constructed by cross-linking two monoclonal antibodies: one specific for tPA, the other specific for fibrin. The bispecific antibody enhanced fibrinolysis by capturing tPA at the site of a fibrin deposit. In an in vitro quantitative fibrinolysis assay, the relative fibrinolytic potency of tPA bound to the bispecific antibody was 13 times greater than that of tPA and 200 times greater than that of urokinase. When fibrin was treated with the bispecific antibody before being mixed with tPA, the relative fibrinolytic potency of tPA was enhanced 14-fold. This capture also occurred when the concentration of tPA present in the assay was less than the concentration of tPA present in normal human plasma. In a human plasma clot assay, samples containing both the bispecific antibody and tPA exhibited significantly more lysis than did samples containing tPA alone. In spite of the increased clot lysis effected by the presence of bispecific antibody, there was no significant increase in fibrinogen or alpha 2-antiplasmin degradation at equal tPA concentrations. The ability of the bispecific antibody to concentrate exogenous tPA in vivo was then examined in the rabbit jugular vein model. Systemic infusion of a small amount of tPA (10,000 units) produced no significant increment in thrombolysis over the level of spontaneous lysis (14 +/- 8%). However, the simultaneous infusion of 10,000 units of tPA and 2 mg of bispecific antibody resulted in 42 +/- 14% (p less than 0.01) lysis. These results suggest that a molecule capable of binding both fibrin and tPA with high affinity could enhance thrombolysis in the circulation by capturing endogenous tPA.
authors.affiliation: Cardiac Unit, Massachusetts General Hospital, Boston.
authors.forename: C
authors.initials: C
authors.lastname: Bode
authors.forename: M S
authors.initials: MS
authors.lastname: Runge
authors.forename: E E
authors.initials: EE
authors.lastname: Branscomb
authors.forename: J B
authors.initials: JB
authors.lastname: Newell
authors.forename: G R
authors.initials: GR
authors.lastname: Matsueda
authors.forename: E
authors.initials: E
authors.lastname: Haber
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000032:analysis; D005337:Fibrin / Q000276:immunology* / Q000502:physiology; D005340:Fibrinogen / Q000032:analysis; D005342:Fibrinolysis; D006801:Humans; D051379:Mice; D008805:Mice, Inbred A; D010958:Plasminogen / Q000032:analysis; D011994:Recombinant Proteins / Q000276:immunology; D010959:Tissue Plasminogen Activator / Q000276:immunology* / Q000502:physiology; D000979:alpha-2-Antiplasmin / Q000032:analysis
chemical_list: D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D011994:Recombinant Proteins; D000979:alpha-2-Antiplasmin; D005337:Fibrin; D005340:Fibrinogen; D010958:Plasminogen; D010959:Tissue Plasminogen Activator