title: Beyond the Natural Proteome: Nondegenerate Saturation Mutagenesis-Methodologies and Advantages.
abstract: Beyond the natural proteome, high-throughput mutagenesis offers the protein engineer an opportunity to "tweak" the wild-type activity of a protein to create a recombinant protein with required attributes. Of the various approaches available, saturation mutagenesis is one of the core techniques employed by protein engineers, and in recent times, nondegenerate saturation mutagenesis is emerging as the approach of choice. This review compares the current methodologies available for conducting nondegenerate saturation mutagenesis with traditional, degenerate saturation and briefly outlines the options available for screening the resulting libraries, to discover a novel protein with the required activity and/or specificity.
authors.affiliation: School of Life and Health Sciences, Aston University, Birmingham, United Kingdom.
authors.forename: M M
authors.initials: MM
authors.lastname: Ferreira Amaral
authors.affiliation: Isogenica Ltd., The Mansion, Chesterford Research Park, Essex, United Kingdom.
authors.forename: L
authors.initials: L
authors.lastname: Frigotto
authors.affiliation: School of Life and Health Sciences, Aston University, Birmingham, United Kingdom. Electronic address: a.v.hine@aston.ac.uk.
authors.forename: A V
authors.initials: AV
authors.lastname: Hine
mesh_terms: D019020:Directed Molecular Evolution; D016296:Mutagenesis; D015202:Protein Engineering; D020543:Proteome / Q000032:analysis*
chemical_list: D020543:Proteome
keywords: Antibody engineering; Directed evolution; Diversity; Gene shuffling molecular evolution; Mutagenesis; Nondegenerate; Protein engineering; Saturation; Synthetic biology