One-step sandwich enzyme-linked immunosorbent assay of human tissue plasminogen activator using monoclonal antibodies.
Five monoclonal antibodies to human tissue plasminogen activator (t-PA) were obtained. Three of them recognized a determinant designated alpha, and other two, another determinant named beta. A t-PA molecule was found to bear one each of determinants alpha and beta arranged in such a manner that the binding of alpha with one of its corresponding antibodies did not interfere with the binding of beta with one of its corresponding antibodies and vice versa. This allowed the development of a one-step sandwich enzyme-linked immunosorbent assay (ELISA) in which the antibody to beta was immobilized on wells of a microtiter plate and the bound t-PA was detected with the enzyme-conjugated antibody to alpha. The ELISA detected both one- and two-chain t-PA with a high sensitivity. We also discuss the determinants, alpha and beta.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000936:Antigen-Antibody Complex; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010954:Plasmacytoma / Q000276:immunology; D010959:Tissue Plasminogen Activator / Q000032:analysis*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D010959:Tissue Plasminogen Activator