TABLE 1.

Experimental and calculated binding affinity for computationally designed point mutants of C225

Mutation	KDa	ΔΔGexpb	ΔΔGcalcc	ΔΔEpaird	ΔΔEHbnde	EHbnd_scf	ΔΔGAbg
	nm	kcal/mol	kcal/mol
WT C225	1.52 ± 0.29	0	0	0	0	0	0
C225-L	NQh		−0.7	−0.4	−0.4	−1.1	1.6
C225-M	15.26	1.36	−0.7	0.1	−0.8	−1.1	6.3
C225-H1	0.77 ± 0.09	−0.41	−2.0	−1.1	−1.2	−2.2	−0.7
C225-H2	1.09 ± 0.13	−0.20	−0.2	−0.1	−0.7	−1.1	3.1
C225-H3	0.37 ± 0.04	−0.84	−0.9	−0.6	−0.2	−0.2	0.4
C225-H	0.08 ± 0.03	−1.75	−3.2				−6.3
F06	5.5
CS06	0.13
B10	24.3
B10v5	0.19

^a The K_D (nm) for wild type (WT) C225 and mutant mAbs was determined by SPR as described under “Experimental Procedures.” Where n > 1, the S.D. is given. K_D values (nm) for c-MET interaction with bivalent anti-c-MET mAbs were determined by BLI. CS06 is the affinity-matured variant of F06, and B10v5 is the derivative of B10.

^b Experimental binding affinity relative to wild type (kcal/mol).

^c Predicted binding affinity relative to wild type using Rosetta.

^d Predicted change in Rosetta pair energy across the interface.

^e Predicted change in hydrogen bond energy across the interface.

^f Calculated hydrogen bond energy of mutated residue side chain.

^g Predicted change in folding energy of the isolated antibody.

^h Not quantifiable; very weak binding.