Protein aggregation and liquid‒liquid phase separation (LLPS) orchestrate protein behavior and function. Engineering protein surface charge offers a robust approach to modulating these phenomena, and supercharging, which replaces surface residues with charged ones, leads to a drastic change in the protein net charge. In this study, we designed a new supercharged antigen-binding fragment antibody mutant and investigated its aggregation behavior. We revealed that the antibody exhibited reversible ligand- and salt concentration-dependent aggregation while retaining the physicochemical properties. Our study demonstrated how supercharging can potentially modulate protein aggregation and LLPS.
- Keisuke Kasahara
- Makoto Nakakido
- Kouhei Tsumoto
