Production of α-Synuclein Fibrillar-Specific scFv from Inclusion Bodies

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 major...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2617; p. 239
Main Authors Gupta, Vijay, Hmila, Issam, Vaikath, Nishant N, Sudhakaran, Indulekha P, El-Agnaf, Omar M A
Format Journal Article
LanguageEnglish
Published United States 2023
Subjects
alpha-Synuclein
Escherichia coli - genetics
Inclusion Bodies
Recombinant Proteins
Single-Chain Antibodies - genetics
Antibody engineering
Protein refolding
scFv
Inclusion bodies
Parkinson’s disease
α-synuclein
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Summary: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.
ISSN:1940-6029
DOI:10.1007/978-1-0716-2930-7_17