TY - JOUR T1 - Modelling of pH-dependence to develop a strategy for stabilising mAbs at acidic steps in production JF - bioRxiv DO - 10.1101/711416 SP - 711416 AU - Max Hebditch AU - Ryan Kean AU - Jim Warwicker Y1 - 2019/01/01 UR - http://biorxiv.org/content/early/2019/07/22/711416.abstract N2 - Engineered proteins are increasingly being required to function or pass through environmental stresses for which the underlying protein has not evolved. A major example in health are antibody therapeutics, where a low pH step is used for purification and viral clearance. In order to develop a computational model for analysis of pH-stability, predictions are compared with experimental data for the relative pH-sensitivities of antibody domains. The model is then applied to proteases that have evolved to be functional in an acid environment, showing a clear signature for low pH-dependence of stability in the neutral to acidic pH region, largely through reduction of saltbridges. Interestingly, an extensively acidic protein surface can maintain contribution to structural stabilisation at acidic pH through replacement of basic sidechains with polar, hydrogen-bonding groups. These observations form a design principle for engineering acid-stable proteins. ER -