Antibody Isolation from Human Synthetic Libraries of Single-Chain Antibodies and Analysis Using NGS.
CDRs (complementarity determining regions)
FR (variable framework region)
HTS (high-throughput sequencing)
NGS (next-generation sequencing
Phage display
Phage-Seq
Single-chain antibodies
Synthetic library
VH (variable region of antibody heavy chain)
VL (variable region of antibody light chain)
scFv (single-chain variable fragment)
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
Antibody libraries came into existence 30 years ago when the accumulating sequence data of immunoglobulin genes and the advent of PCR technology made it possible to clone antibody gene repertoires. Phage display (most common) and additional display and screening technologies were applied to pan out desired binding specificities from antibody libraries. As other antibody discovery tools, phage display is not an off-the-shelf technology and not offered as a kit but rather requires experience and expertise for making it indeed very useful.Next-generation sequencing (NGS) coupled with bioinformatics is a powerful tool for analyzing large amount of DNA sequence output of the panning. Here, we demonstrate how NGS analysis of phage biopanning (phage-Seq) of complex antibody libraries can facilitate the antibody discovery process and provide insights regarding the biopanning process (see Fig. 1).
Identifiants
pubmed: 37679629
doi: 10.1007/978-1-0716-3381-6_18
doi:
Substances chimiques
Single-Chain Antibodies
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
347-372Informations de copyright
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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