Beyond the natural proteome: nondegenerate saturation mutagenesis - methodologies and advantages

M.M. Ferreira Amaral; L. Frigotto; A.V. Hine

doi:10.1016/bs.mie.2016.10.005

Beyond the natural proteome: nondegenerate saturation mutagenesis - methodologies and advantages

M.M. Ferreira Amaral, L. Frigotto, A.V. Hine^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Pages (from-to)	111-133
Journal	Methods in Enzymology
Volume	585
DOIs	https://doi.org/10.1016/bs.mie.2016.10.005
Publication status	Published - 3 Jan 2017

Bibliographical note

Proteomics in biology, part A ISBN: 978-0-1280-9742-7.
© 2016, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/

Keywords

gene shuffling
molecular evolution
directed evolution
synthetic biology
saturation mutagenesis
non-degenerate
diversity
protein engineering
antibody engineering

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10.1016/bs.mie.2016.10.005

Beyond the natural proteome
© 2016, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/
Accepted author manuscript, 686 KBLicence: CC BY-NC-ND 3.0

Cite this

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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. ",

keywords = "gene shuffling, molecular evolution, directed evolution, synthetic biology, saturation mutagenesis, non-degenerate, diversity, protein engineering, antibody engineering",

author = "{Ferreira Amaral}, M.M. and L. Frigotto and A.V. Hine",

note = "Proteomics in biology, part A ISBN: 978-0-1280-9742-7. {\textcopyright} 2016, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ ",

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AU - Hine, A.V.

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Beyond the natural proteome: nondegenerate saturation mutagenesis - methodologies and advantages

Abstract

Bibliographical note

Keywords

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