Construction of Semisynthetic Shark vNAR Yeast Surface Display Antibody Libraries.
Affinity maturation
Antibody engineering
IgNAR
Library generation
Protein engineering
Semisynthetic antibody library
Shark
Single domain antibody
Yeast surface display
vNAR
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:
2023
2023
Historique:
medline:
11
9
2023
pubmed:
8
9
2023
entrez:
7
9
2023
Statut:
ppublish
Résumé
The adaptive immune system of sharks comprises a unique heavy chain-only antibody isotype, termed immunoglobulin new antigen receptor (IgNAR), in which antigen binding is mediated by a single variable domain, referred to as vNAR. In recent years, efforts were made to harness these domains for biomedical and biotechnological applications particularly due to their high affinity and specificity combined with a small size and high stability. Herein, we describe protocols for the construction of semisynthetic, CDR3-randomized vNAR libraries for the isolation of target-specific paratopes by yeast surface display. Additionally, we provide guidance for affinity maturation of a panel of antigen-enriched vNAR domains through CDR1 diversification of the FACS-selected, antigen-enriched population and sublibrary establishment.
Identifiants
pubmed: 37679622
doi: 10.1007/978-1-0716-3381-6_11
doi:
Substances chimiques
Antibodies
0
Receptors, Antigen, B-Cell
0
Antibodies, Fungal
0
Immunoglobulin Isotypes
0
Immunoglobulin Heavy Chains
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
227-243Informations de copyright
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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