Assessing and Engineering Antibody Stability Using Experimental and Computational Methods

Abstract

Engineering increased stability into antibodies can improve their developability. While a range of properties need to be optimized, thermal stability and aggregation are two key factors that affect the antibody yield, purity, and specificity throughout the development and manufacturing pipeline. Therefore, an ideal goal would be to apply protein engineering methods early-on, such as in parallel to affinity maturation, to screen out potential drug molecules with the desired conformational and colloidal stability. This chapter introduces our methods to computationally characterize an antibody Fab fragment, propose stabilizing variants, and then experimentally verify these predictions.

Keywords: Aggregation; Antibody; B-factor; Entropy; Fab; Molecular dynamics (Gromacs); Rosetta; Tm.