Facile isolation of high-affinity nanobodies from synthetic libraries using CDR-swapping mutagenesis.
STAR Protoc
; 3(1): 101101, 2022 03 18.
Article
in English
| MEDLINE | ID: covidwho-1626432
ABSTRACT
The generation of high-affinity nanobodies for diverse biomedical applications typically requires immunization or affinity maturation. Here, we report a simple protocol using complementarity-determining region (CDR)-swapping mutagenesis to isolate high-affinity nanobodies from common framework libraries. This approach involves shuffling the CDRs of low-affinity variants during the sorting of yeast-displayed libraries to directly isolate high-affinity nanobodies without the need for lead isolation and optimization. We expect this approach, which we demonstrate for SARS-CoV-2 neutralizing nanobodies, will simplify the generation of high-affinity nanobodies. For complete details on the use and execution of this profile, please refer to Zupancic et al. (2021).
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Single-Domain Antibodies
/
COVID-19
Type of study:
Randomized controlled trials
Topics:
Variants
Limits:
Humans
Language:
English
Journal:
STAR Protoc
Year:
2022
Document Type:
Article
Affiliation country:
J.xpro.2021.101101
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