Table 2.

Thermodynamic constants for the binding of MIP NPs to different analytes measured by Isothermal Titration Calorimetry (ITC)

NP	Ligand	Kd (nM)	n	ΔH (kJ/mol)	ΔG (kJ/mol)
02 MIP32	DTHFPI	6 ± 3	0.4 ± 0.1	−261 ± 18	−47 ± 2
02 MIP200	DTHFPI	7 ± 1	0.4 ± 0.1	−128 ± 25	−46 ± 1
05 MIP32	DTHFPI	3 ± 1	0.4 ± 0.1	−177 ± 14	−49 ± 1
05 MIP200	DTHFPI	5 ± 3	0.5 ± 0.1	−131 ± 4	−48 ± 2
05 MIP200	Hepcidin-25	13 ± 2	0.3 ± 0.1	−463 ± 46	−51 ± 9
05 MIP200	Hepcidin-20	–	–	–	–
05 MIP200	THFDPI	19 ± 5	0.5 ± 0.1	−395 ± 27	−37 ± 2
05 MIP200	NR10	–	–	–	–
02 NIP	DTHFPI	–	–	–	–
05 NIP	DTHFPI	–	–	–	–
05 NIP	Hepcidin-25	–	–	–	–

Experiments were performed by titrating 1.2 µM NPs with 4 µM ligand: dissociation constant (Kd), binding stoichiometry (n), enthalpy and Gibbs free energy variation. The heat contribution of control NPs was subtracted from MIP NPs, thus the resulting interactions are primarily driven by specific binding sites. Thermograms in Additional file 1