Production of α-Synuclein Fibrillar-Specific scFv from Inclusion Bodies.
Antibody engineering
Inclusion bodies
Parkinson’s disease
Protein refolding
scFv
α-synuclein
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:
entrez:
19
1
2023
pubmed:
20
1
2023
medline:
24
1
2023
Statut:
ppublish
Résumé
Recombinant antibody fragments such as Fab, scFvs, and diabodies against α-syn have become a viable alternative to the conventional full-length antibodies in immunotherapeutic approaches due to their benefits which include smaller size, higher stability, specificity, and affinity. However, the majority of recombinant antibody fragments typically express as inclusion bodies (IBs) in E. coli, which makes their purification incredibly difficult. Here, we describe a method involving a mild solubilizing protocol followed by slow on-column refolding to purify active single-chain variable fragment (scFv-pF) antibody that can recognize the pathogenic α-syn fibrils.
Identifiants
pubmed: 36656529
doi: 10.1007/978-1-0716-2930-7_17
doi:
Substances chimiques
alpha-Synuclein
0
Single-Chain Antibodies
0
Recombinant Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
239-248Informations de copyright
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
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