Nanobodies as solubilization chaperones for the expression and purification of inclusion-body prone proteins

Here, we report a new protocol for enhancing the soluble expression of inclusion body (IB)-prone proteins in E. coli using nanobodies (Nbs) as a molecular-specific chaperone. The specific intracellular binding between the cognate-Nbs and the antigen is successfully achieved and enables the formation...

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Bibliographic Details
Published in:Chemical communications (Cambridge, England) England), 2022-02, Vol.58 (17), p.2898-291
Main Authors: Yao, Guangshuai, Huang, Chundong, Ji, Fangling, Ren, Jun, Luo, Xiuna, Zang, Berlin, Jia, Lingyun
Format: Article
Language:English
Subjects:
Antigens
Binding
E coli
Escherichia coli Proteins - chemistry
Inclusion Bodies - chemistry
Molecular Chaperones - chemistry
Proteins
Single-Domain Antibodies - chemistry
Solubility
Solubilization
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Summary:Here, we report a new protocol for enhancing the soluble expression of inclusion body (IB)-prone proteins in E. coli using nanobodies (Nbs) as a molecular-specific chaperone. The specific intracellular binding between the cognate-Nbs and the antigen is successfully achieved and enables the formation of a soluble Nb-antigen complex in E. coli . We further expand this method by adding an epitope tag (EPEA-tag) to the target proteins, and the anti-EPEA Nb was intended to act as the chaperone for in vivo binding with the EPEA tag. Such substitution may develop a "multi-specific" Nb-chaperone that can simultaneously and effectively cope with different IB proteins of interest. Nanobodies as molecular-specific chaperones assist in the folding of antigen proteins via specific binding in vivo.
ISSN:1359-7345
1364-548X
DOI:10.1039/d1cc07105j