Application of Deep Sequencing in Phage Display.

Enrichment detection Illumina sequencing Indel detection Molecular evolution Next-generation sequencing Oxford Nanopore sequencing Phage 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:
2024
Historique:
medline: 16 11 2023
pubmed: 15 11 2023
entrez: 15 11 2023
Statut: ppublish

Résumé

This chapter describes the workflow to implement deep sequencing into standard phage display experiments on protein libraries. By harvesting the power of high throughput of these techniques, it allows for comprehensive analysis of the naïve library and library evolution in response to selection by ligand binding. The mutagenized target region of the protein variants encoded by the phage pool is analyzed by Illumina paired-end sequencing. Sequence data are processed to extract selection-enriched amino acid motifs. In addition, a complementary long-read sequencing approach is proposed enabling the monitoring of display vector stability.

Identifiants

pubmed: 37966608
doi: 10.1007/978-1-0716-3549-0_20
doi:

Substances chimiques

Mutant Proteins 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

333-345

Informations de copyright

© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

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Auteurs

Vincent Van Deuren (V)

Department of Biochemistry, Molecular and Structural Biology, KU Leuven, Leuven, Belgium.

Sander Plessers (S)

Department of Biochemistry, Molecular and Structural Biology, KU Leuven, Leuven, Belgium.

Rob Lavigne (R)

Animal and Human Health Engineering (A2H), Leuven (Arenberg), KU Leuven, Leuven, Belgium.

Johan Robben (J)

Department of Biochemistry, Molecular and Structural Biology, KU Leuven, Leuven, Belgium. Johan.robben@kuleuven.be.

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Classifications MeSH