Engineering Proteins by Combining Deep Mutational Scanning and Yeast Display.

Deep mutational scanning Enrichment ratio Heat maps Protein engineering Sequence-activity landscape T cell receptors Yeast display

Journal

Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969

Informations de publication

Date de publication:
2022
Historique:
entrez: 28 4 2022
pubmed: 29 4 2022
medline: 3 5 2022
Statut: ppublish

Résumé

Protein engineering using display platforms such as yeast display and phage display has allowed discovery of proteins with therapeutic and industrial applications. Antibodies and T cell receptors developed for therapeutic applications are often engineered by constructing libraries of mutations in loops of five to ten residues called complementarity determining regions that are in proximity to the antigen. In the past decade, deep mutational scanning has become a powerful tool in a protein engineer's toolbox, as it allows one to compare the impact of all 20 amino acids at each position, across the length of the protein. Thus, a single experiment can provide a sequence-activity landscape with information about hotspots or suboptimal binding sites in the original proteins. These residues or regions may be overlooked by engineering methods that are driven solely by structures or directed evolution of error-prone PCR libraries. Here, we describe experimental methods to engineer proteins by combining yeast display and deep mutational scanning mutagenesis, using T cell receptors as an example.

Identifiants

pubmed: 35482188
doi: 10.1007/978-1-0716-2285-8_7
doi:

Substances chimiques

Receptors, Antigen, T-Cell 0

Types de publication

Journal Article Research Support, N.I.H., Extramural

Langues

eng

Sous-ensembles de citation

IM

Pagination

117-142

Subventions

Organisme : NCI NIH HHS
ID : R21 CA238628
Pays : United States

Informations de copyright

© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

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Auteurs

Preeti Sharma (P)

Department of Biochemistry, University of Illinois, Urbana, IL, USA.
Cancer Center at Illinois, University of Illinois, Urbana, IL, USA.

Erik Procko (E)

Department of Biochemistry, University of Illinois, Urbana, IL, USA.
Cancer Center at Illinois, University of Illinois, Urbana, IL, USA.

David M Kranz (DM)

Department of Biochemistry, University of Illinois, Urbana, IL, USA. d-kranz@illinois.edu.
Cancer Center at Illinois, University of Illinois, Urbana, IL, USA. d-kranz@illinois.edu.

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