A one-step enzyme immunoassay for the determination of total tissue-type plasminogen activator (t-PA) antigen in plasma.
A reproducible and sensitive one-step enzyme immunoassay (EIA) was developed to determine total tissue-type plasminogen activator (t-PA) antigen in plasma. The EIA comprises two monoclonal catching antibodies and a polyclonal (goat) tagging antibody conjugated with horseradish peroxidase. There is an equal reactivity towards the several physiological t-PA forms, i.e., single-chain t-PA, two-chain t-PA and t-PA in complex with its naturally occurring inhibitor plasminogen activator inhibitor-type 1 (t-PA/PAI-1 complex). Additionally, the EIA does not discriminate between human melanoma t-PA and recombinant t-PA (Activase). The assay has a lower detection limit of approximately 0.5 ng t-PA per ml plasma, with a time-to-result of only 3.5 h.
Bos
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R
IVVO-TNO Gaubius Laboratory, Leiden, The Netherlands.
Hoegee-de Nobel
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E
Laterveer
R
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Meyer
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P
Nieuwenhuizen
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W
D000906:Antibodies; D000911:Antibodies, Monoclonal / Q000276:immunology; D006735:Horseradish Peroxidase; D007124:Immunoenzyme Techniques; D010949:Plasma / Q000737:chemistry; D015849:Plasminogen Inactivators; D011994:Recombinant Proteins / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D010959:Tissue Plasminogen Activator / Q000032:analysis* / Q000276:immunology
D000906:Antibodies; D000911:Antibodies, Monoclonal; D015849:Plasminogen Inactivators; D011994:Recombinant Proteins; D006735:Horseradish Peroxidase; D010959:Tissue Plasminogen Activator