| Parameter | Description | Fitting status |
|---|---|---|
| L | Concentration of free ligand (RU) | Constant |
| A | Concentration of free analyte (M) | Constant |
| LA | Concentration of ligand-analyte complex (RU) | Constant |
| LAA | Concentration of two ligand-analyte complex (RU) | Fit globally |
| ka1 | Association rate constant for A + 2L ⇌ LA (M-1s-1) | Fit globally |
| kd1 | Dissociation rate constant for LA⇌ 2L + A (s-1) | Fit globally |
| ka2 | Association rate constant for LA + L ⇌ LLA (RU-1s-1) | Fit globally |
| kd2 | Dissociation rate constant for LLA ⇌ LA + L (s-1) | Fit globally |
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