A solid-phase fibrin immunoassay for the specific detection of monoclonal antibodies against different epitopic determinants of tissue-plasminogen activators.
The selective detection of hybridomas producing monoclonal antibodies against tissue-plasminogen activators (t-PA) by means of a solid-phase fibrin immunoassay (SOFIA) is described. The screening method is based on the specific and high affinity binding of t-PA for fibrin solid phase. This, as in the physiological fibrin-t-PA interaction, provides a molecular structure conserving its antigenic and functional properties. The solid-fibrin-state support is prepared by thrombin proteolysis of fibrinogen coupled to a poly-glutaraldehyde activated polyvinylchloride plate. This procedure avoids fibrinogen desorption and provides a uniform fibrin network onto which t-PA from crude extracts or purified preparations, can be specifically absorbed. The fibrin-t-PA solid-phase support is able to fix anti-t-PA antibodies which can be subsequently revealed by a second enzyme- or radiolabelled anti-mouse Ig antibody. The results demonstrate the effectiveness of the SOFIA method for the selection of hybridomas producing anti-t-PA antibodies against different epitopic determinants of t-PAs.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D000939:Epitopes / Q000276:immunology*; D005337:Fibrin; D005340:Fibrinogen / Q000378:metabolism; D005976:Glutaral; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D010960:Plasminogen Activators / Q000276:immunology* / Q000378:metabolism; D011143:Polyvinyl Chloride; D013552:Swine
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005337:Fibrin; D005340:Fibrinogen; D011143:Polyvinyl Chloride; D010960:Plasminogen Activators; D005976:Glutaral