Table 6.
Summary of the effect of single point mutations on thermal stability, solubility, viscosity and biological activity of MAB 1*
| Mutation, location and solvent exposure | Thermal stability (DSC) | Apparent solubility | DLS self-association tendency | Viscosity | Biological activity | Other observations |
|---|---|---|---|---|---|---|
| M2 (E59R), FR3 in VL, solvent exposed | All Tms lowered | Less than M1 and MAB 1 Control | Greater than M1 and MAB 1 Control | Not enough data | Active | Harder to concentrate, increased opalescence and phase separation |
| M3 (E59Y), FR3 in VL, solvent exposed | Lower Tm1 | Greater than M1 and MAB 1 Control | Less than M1 and MAB 1 Control | Reduced viscosity | Active | Improved overall solution behavior |
| M4 (E59K), FR3 in VL, solvent exposed | Lower Tm1 and Tm2 | Less than M1 and MAB 1 Control | Similar to M1 but less than MAB 1 Control | Not enough data | Active | Harder to concentrate, increased opalescence and phase separation |
| M5 (L45K), FR2 in VL, lies at VH:VL interface, solvent inaccessible | Atypical DSC curve | n. d. | n. d. | n. d. | Inactive | Significantly destabilized |
| M6 (V44K), FR2 in VL, solvent exposed | All Tms similar to M1 and MAB 1 Control | Greater than M1 and MAB 1 Control | Greater than M1 but less than MAB 1 Control | Reduced viscosity | Active | Improved overall solution behavior |
*
n. d. stands for not determined. Bioactivity indicates antigen binding as measured by in vitro competitive binding ELISA assays.