An enzyme immunoassay for the simultaneous determination of active type-1 plasminogen activator inhibitor (PAI-1), and t-PA/PAI-1 complexes.
An assay has been developed which enables the simultaneous determination of active free PAI-1 and t-PA/PAI-1 complexes in plasma samples. Test plasma is divided into two subsamples in which (a) active PAI-1 is quantitatively converted to t-PA/PAI-1 complexes, and (b) conversion of PAI-1 to t-PA/PAI-1 complexes is prevented. The amounts of t-PA/PAI-1 complex in the two subsamples are then assessed in a one-step enzyme immunoassay (EIA) consisting of a monoclonal anti-PA antibody, and a HRP conjugate of polyclonal anti-PAI-1 antibodies. The amounts of t-PA/PAI-1 complexes in the two subsamples are the t-PA/PAI-1 complex concentrations at the time of sampling (b); whereas the difference between the two values (a)-(b) equals the amount of active PAI-1. The assay has a high sensitivity, specificity, accuracy and precision. The time-to-result is 4.5 h. It meets the criteria set by the Scientific and Standardization Committee (SSC) of the International Society of Thrombosis and Haemostasis (ISTH) for a candidate reference method.
Nieuwenhuizen
W
W
Gaubius Laboratory TNO-PG, Leiden, The Netherlands.
Laterveer
R
R
Hoegee
B
B
D002138:Calibration; D004355:Drug Stability; D006801:Humans; D007124:Immunoenzyme Techniques / Q000706:statistics & numerical data; D017395:Plasminogen Activator Inhibitor 1 / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D010959:Tissue Plasminogen Activator / Q000097:blood*
D017395:Plasminogen Activator Inhibitor 1; D010959:Tissue Plasminogen Activator