Table 2.
Overview of the existing plasmin generation assays and their major properties and applications.
| NHA | STA | STPGA | PGA | |
|---|---|---|---|---|
| Substrates (thrombin and plasmin) | Bz-AGR-AMC and bis-(CBZ-FR)-Rho |
Boc-VPR-AMC and Boc-EKK-AMC |
Z-GGR-AMC and Boc-EKK-AMC |
Boc-EKK-AMC |
| Specificity of the plasmin substrate | Plasmin, thrombin, factor Xa |
Plasmin | Plasmin | Plasmin |
| Calibration method | Calibration with plasmin | - | Michaelis–Menten equation using the obtained values of AMC production rate | Calibration with α2M-plasmin |
| Method to obtain plasmin curve | Comparing the arbitrary fluorescence values to a calibration curve | Subtracting the fluorescence reading of the blank well | Plasmin is calculated using the obtained values of AMC production rate | Comparing the arbitrary fluorescence values to a calibration curve |
| Correction for substrate consumption/inner filter effect | +/+ | −/− | −/+ | +/+ |
| tPA (μg/mL) | 0.38 | 0.45 | 0.7 | 0.31 or 1.25 |
| TF (pM) | 0.3 | 5 | 4.5 | 0.5 or 1 |
| Plasma volume | 80 | 90 | 80 | 30 |
| Published applications in health and diseases | -PG in patients with plasminogen and PAI-1 deficiency [92] -PG in hemophilia A patients [93] -PG in patients with rare bleeding disorders [81] -PG in patients with angioedema [82] |
-Abnormalities in fibrinolysis in adults and children [83] -PG in human plasma with fibrinolytic deficiency [84] |
-Inhibitory effect of C-1 inhibitor [85] | -PG in mice model of diet-induced obesity [44] -PG in mice with fibrin(ogen) deficiency [44] -Pharmacodynamics of TXA [88] -PG in coronavirus disease-2019 patients [94,95] |
NHA (Nijmegen hemostasis assay), STA (simultaneous thrombin and plasmin generation assay), STPGA (simultaneous thrombin plasmin generation assay), PGA (plasmin generation assay). Boc-EKK-AMC (Boc-Glu-Lys-Lys-AMC), bis-(CBZ-FR)-Rho (bis-(CBZ-L-phenylalanyl-L-arginine amide)-rhodamine); α2M-plasmin (α2-Macroglobulin-plasmin).