Immunotargeting of erythrocyte-bound streptokinase provides local lysis of a fibrin clot.
The creation of an anticollagen antibody-erythrocyte-streptokinase complex has been described. Immobilization of both proteins on erythrocyte membrane has been performed using an avidin-biotin interaction. Modification of streptokinase with (6-biotinylamido)hexanoic acid N-hydroxysuccinimide ester at the concentration of 1.1 mM (20% modification of protein amino groups) provides effective (up to 90%) attachment of streptokinase to an avidin-carrying erythrocyte surface. The loss of streptokinase activity due to modification under these conditions is not significant. The maximal attachment of streptokinase was equal to about 50 ng per 10(6) erythrocytes, i.e., about 5 X 10(5) molecules of streptokinase per erythrocyte. The presence of streptokinase in the incubation mixture inhibited the attachment of antibodies by about 50%. Nevertheless, co-immobilization of anticollagen antibody (1.0 X 10(5) molecules per cell) and streptokinase (2.8 X 10(5) molecules per cell) on the erythrocyte surface provided firm and specific binding of such erythrocytes to a collagen-coated surface (1.6 X 10(6) bound cells per 1 cm2 on a collagen-coated surface against 0.006 X 10(6) bound cells on a bovine serum albumin-coated surface). Targeting of such erythrocytes led to local lysis of a fibrin clot in the target zone. The properties described offer in principle the possibility of the application of this or a similar system of fibrinolytic agent targeting for the preventive therapy of rethrombosis during surgical manipulations on vessels.
D000906:Antibodies; D001710:Biotin / Q000031:analogs & derivatives; D003094:Collagen / Q000276:immunology; D004800:Enzymes, Immobilized / Q000378:metabolism*; D004910:Erythrocyte Membrane; D005342:Fibrinolysis; D006801:Humans; D011485:Protein Binding; D013300:Streptokinase / Q000008:administration & dosage / Q000097:blood*; D013388:Succinimides
D000906:Antibodies; D004800:Enzymes, Immobilized; D013388:Succinimides; C033788:biotinyl N-hydroxysuccinimide ester; D001710:Biotin; D003094:Collagen; D013300:Streptokinase