Beyond the natural proteome

nondegenerate saturation mutagenesis - methodologies and advantages

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

Research output: Contribution to journalArticle

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 languageEnglish
JournalMethods in Enzymology
Early online date3 Jan 2017
DOIs
Publication statusE-pub ahead of print - 3 Jan 2017

Fingerprint

Mutagenesis
Proteome
Proteins
Engineers
Recombinant Proteins
Libraries
Screening
Throughput

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

Cite this

@article{5662a3ad626240f19eb8cab394ea0cff,
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. {\circledC} 2016, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/",
year = "2017",
month = "1",
day = "3",
doi = "10.1016/bs.mie.2016.10.005",
language = "English",
journal = "Methods in Enzymology",
issn = "0076-6879",
publisher = "Academic Press Inc.",

}

Beyond the natural proteome : nondegenerate saturation mutagenesis - methodologies and advantages. / Ferreira Amaral, M.M.; Frigotto, L.; Hine, A.V.

In: Methods in Enzymology, 03.01.2017.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Beyond the natural proteome

T2 - nondegenerate saturation mutagenesis - methodologies and advantages

AU - Ferreira Amaral, M.M.

AU - Frigotto, L.

AU - Hine, A.V.

N1 - 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/

PY - 2017/1/3

Y1 - 2017/1/3

N2 - 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.

AB - 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.

KW - gene shuffling

KW - molecular evolution

KW - directed evolution

KW - synthetic biology

KW - saturation mutagenesis

KW - non-degenerate

KW - diversity

KW - protein engineering

KW - antibody engineering

UR - http://www.scopus.com/inward/record.url?scp=85008324794&partnerID=8YFLogxK

U2 - 10.1016/bs.mie.2016.10.005

DO - 10.1016/bs.mie.2016.10.005

M3 - Article

JO - Methods in Enzymology

JF - Methods in Enzymology

SN - 0076-6879

ER -