Fig. 1

Using library-based display technologies to evolve protein stability. Genetic diversification of the POI can be achieved through various strategies such as error-prone PCR, DNA shuffling, and incorporation of unnatural amino acid. The library can be displayed on various surfaces of carriers, such as yeast, phage, and spore, or conjugated with ribosome and mRNA. Protein unfolding pressures are applied to eliminate the unstable variants from the POI library. Stabilized variants are obtained based on their intrinsic properties, such as ligand binding. The selected or screened variants are sequenced, verified, and ready to enter a new round of evolution