Enzyme-linked immunosorbent assay for human urokinase-type plasminogen activator and its proenzyme using a combination of monoclonal and polyclonal antibodies.
A sensitive and specific enzyme-linked immunosorbent assay (ELISA) for human urokinase-type plasminogen activator (u-PA) and its inactive proenzyme (pro-u-PA) was developed. A monoclonal antibody was used as solid-phase antibody, while rabbit antibodies against human u-PA followed by peroxidase-conjugated third antibody were used for detection of bound u-PA. No reaction was observed with tissue-type plasminogen activator or with a variety of other human proteins. The assay was used for quantitation of u-PA in human urine and in culture fluid from human tumor cells. The recovery of added pro-u-PA was greater than 95%. A good agreement with the results obtained by enzymatic assays was found. The detection limit was less than 0.1 ng per ml, both for u-PA and pro-u-PA. The advantages of the use of ELISA compared with enzymatic assays and radioimmunoassays for quantitation of u-PA and pro-u-PA in biological samples are discussed.
D000906:Antibodies; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D010960:Plasminogen Activators / Q000032:analysis* / Q000276:immunology; D011863:Radioimmunoassay; D014568:Urokinase-Type Plasminogen Activator / Q000032:analysis* / Q000276:immunology
D000906:Antibodies; D000911:Antibodies, Monoclonal; D010960:Plasminogen Activators; D014568:Urokinase-Type Plasminogen Activator