Non-native protein aggregation is an outstanding issue in the biopharmaceutical industry, making rational design of aggregation-resistant proteins and conditions an important goal. Different approaches have been used previously, including �supercharging� of the surface of a protein to maximize repulsive electrostatic interactions. For this work, we are examining the effects of single point mutations that alter the surface charge on human gamma-D crystallin, and are predicted to modulate protein monomer interactions without having a substantial impact on folding stability. Our models suggest that these modifications alter the protein's aggregation propensity and provide insight into the effects of monomer-monomer interactions on protein aggregation. Our experimental observations indicate that even single-charge-altering point mutations have significant effects on aggregation rates, with complementary techniques (e.g., light scattering) used to investigate the mechanism(s) behind these changes. Ongoing work extends this approach to additional molecules, such as single chain antibodies.

Combining theory and experiment for rational design of single-charge-altering point mutations to reduce multi-domain protein aggregation

245th National Meeting (2013)

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