. 2014 Sep 5;171(23):5295–5312. doi: 10.1111/bph.12836

Table 6.

Binding of the monovalent ligand ‘c’ (k₊_c = 1 × 10⁷ M⁻¹·min⁻¹, k₋_c = 0.3 min⁻¹ and [c] = 100 nM) was simulated during a time lapse of 50 min in the absence or presence of different concentrations of the bitopic ligands ‘ab’ (k₊_a = 1 × 10⁵ M⁻¹·min⁻¹, k₋_a = 1 min⁻¹, k₊_b = 1 × 10⁵ M⁻¹·min⁻¹, k₋_b = 1 min⁻¹ and [L] = 0.29 mM) with different cooperativity factors

Example	Cooperativity factor	Calculated parameters^a		Collected parameters^b
Example	Cooperativity factor	k_on (M⁻¹·min⁻¹)	k_off (min⁻¹)	k_on (M⁻¹·min⁻¹)	k_off (min⁻¹)
1	α_ab = 10	2.0 × 10⁵	0.0069	2.3 × 10⁵	0.0046
2	α_ab = 0.1	1.6 × 10⁵	0.41	1.1 × 10⁵	0.72
3	α'_ab = 10	1.9 × 10⁵	0.0066	2.1 × 10⁵	0.0039
4	α'_ab = 0.1	2.0 × 10⁵	0.50	1.3 × 10⁵	0.90

The ‘macroscopic’ kinetic binding parameters of the bitopic ligands were calculated using Equations 13 and 14 or by subjecting the simulated data (Supporting Information Tables S1–S4) into the competition association assay based on the theoretical framework by Motulsky and Mahan (1984).

The ‘macroscopic’ kinetic binding parameters of the bitopic ligands were calculated using Equations 13 and 14.

k_on and k_off values were determined by analysing the simulated data (Supporting Information Tables S1–S4) in the competition association model ‘kinetics of competitive binding’ (Motulsky and Mahan, 1984).