IMUBIND® Factor VIIa ELISA

The IMUBIND FVIIa ELISA assay uses a biotinylated FVIIa enzyme inhibitor and anti-FVII / FVIIa monoclonal antibody as the capture antibody.
Diluted plasma samples or supernatants containing FVIIa are incubated with the biotinylated inhibitor, which covalently binds to FVIIa but not FVII.
The samples are added to the microwell coated with the capture monoclonal antibody. The FVIIa is detected thanks to the streptavidin-HRP which will bind the FVIIa complex captured at the bottom of the well by the monoclonal antibody and the biotinylated FVIIa inhibitor.
The TMB will thus recognize the HRP giving a blue compound which will be stopped by adding sulfuric acid giving a yellow compound, measured at 450nm. The results will be compared with a known FVIIa standard curve.

Factor VII (FVII) is a glycoprotein synthesized by the liver, vitamin k dependent. When tissue factor (TF) appears on the surface of damaged, abnormal or activated vascular endothelium, FVIIa associates with it, initiating the pathway extrinsic coagulation.
The FT-FVIIa complex activates the FX in FXa and the FIX in FIXa.