TABLE 2 .
Thermodynamic parameters of the interaction of MN86-994-11 with rP2086-B01 epitope mutantsa
| Mutation | Ka (M−1) | Kd (nM) | ΔH (kcal/mol) | ΔS ([cal/mol] deg−1) |
|---|---|---|---|---|
| None (wild type) | (1.8 ± 0.2) × 108 | 6 | −11.9 ± 0.0 | −2.2 |
| K185A | (0.9 ± 0.1) × 108 | 11 | −10.8 ± 0.1 | 0.3 |
| E187A | (1.2 ± 0.2) × 108 | 8 | −10.7 ± 0.1 | 1.0 |
| K190A | (5.9 ± 0.4) × 105 | 1,700 | −9.3 ± 0.2 | −4.9 |
| S191A | (1.1 ± 0.2) × 107 | 89 | −8.6 ± 0.1 | 3.5 |
| N195A | (4.7 ± 0.4) × 105 | 2,100 | −8.9 ± 0.3 | −3.8 |
| D197A | (6.9 ± 1.5) × 107 | 14 | −12.4 ± 0.1 | −5.9 |
a
Alanine substitution mutations were introduced into conserved amino acid residues located within the epitope predicted by HDX-MS mapping (Fig. 2 and 3). Residues were targeted on the basis of their orientation and potential for forming hydrogen bonds or salt bridges with the antibody. The parameters were derived from the ITC titrations of the antibody with the wild-type and mutant proteins, as described in the text. Data are from single experiments with each protein. The errors shown are errors of fit.