Co-expression of Dsb proteins enables soluble expression of a single-chain variable fragment (scFv) against human type 1 insulin-like growth factor receptor (IGF-1R) in E. coli
Abstract
Type 1 insulin-like growth factor receptor (IGF-1R) is a promising therapeutic target for cancer treatment. A single-chain variable fragment (scFv) against human IGF-1R forms inclusion body when expressed in periplasmic space of E. coli routinely. Here, we described that co-expression of appropriate disulfide bonds (Dsb) proteins known to catalyze the formation and isomerization of Dsb can markedly recover the soluble expression of target scFv in E. coli. A 50 % recovery in solubility of the scFv was observed upon co-expression of DsbC alone, and a maximum solubility (80 %) was obtained when DsbA and DsbC were co-expressed in combination. Furthermore, the soluble scFv present full antigen-binding activity with IGF-1R, suggesting its correct folding. This study also suggested that the selection of Dsb proteins should be tested case-by-case if the approach of co-expression of Dsb system is adopted to address the problem of insoluble expression of proteins carrying Dsb.
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Sun reported a yield of 33 mg/L of a soluble and active scFv in shaking-flask cultures using BL21 (DE3) cells, 2xYT media and induction with 0.2 mM IPTG for 4 h at 30 °C.
“Sun and coworkers reported a yield of 33 mg/L of a soluble and active scFv in shaking-flask cultures by using BL21 (DE3) cells, 2xYT media and following induction with 0.2 mM IPTG for 4 h at 30 °C [170].”
From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [170]
The synergistic effect of DsbA/DsbC has been successfully assessed as an effective way to improve the soluble expression of Fabs and scFvs in the E. coli periplasmic space.
“The synergistic effect of DsbA/DsbC has been next successfully assessed as an effective way to improve the soluble expression of Fabs [56,88] and scFvs [170] in the E. coli periplasmic space.”
From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [170]
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Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments
Sandomenico, A., Sivaccumar, J. P., & Ruvo, M. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21176324 · Cited by 134 · see citing works
- Authors
- Sun, X.-W., Wang, X.-H., & Yao, Y.-B.
- Affiliations
- Hebei University of Engineering (CN)
- Venue
- World Journal of Microbiology and Biotechnology
- Published
- 2014-09-26
- Volume
- 30
- Issue
- 12
- Pages
- 3221-3227
- ISSN
- 0959-3993 (ISSN-L)
- 1573-0972
- OpenAlex
- W1965607811
- Cited by
- 12
- References
- 0
- Citations by year
- 2026: 1 · 2021: 1 · 2020: 3 · 2019: 1 · 2018: 3 · 2017: 1 · 2015: 2
- FWCI
- 0.41
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- World Journal of Microbiology and Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- DsbA, DsbC, scFv, IGF-1 receptor, soluble protein expression, inclusion bodies, protein folding, antigen binding activity, recombinant antibody fragments, Escherichia coli
- MeSH terms
- Biotechnology, Escherichia coli, Humans, Protein Binding, Recombinant Proteins, Solubility, Receptors, Somatomedin, Protein Folding, Protein Disulfide-Isomerases, Escherichia coli Proteins, Single-Chain Antibodies
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