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Cited by 62Monoclonal and Polyclonal Antibodies ResearchProtein purification and stabilityBiochemical and Structural Characterization

Engineering production of functional scFv antibody in E. coli by co-expressing the molecule chaperone Skp

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Wang, Rongzhi, Xiang, Shuangshuang, Feng, Youjun, Srinivas, Swaminath, Zhang, Yonghui, Lin, Mingshen, Wang, Shihua·2013·Frontiers in Cellular and Infection Microbiology
DOI 10.3389/fcimb.2013.00072PMC PMC3818579
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Abstract

Single-chain variable fragment (scFv) is a class of engineered antibodies generated by the fusion of the heavy (VH) and light chains (VL) of immunoglobulins through a short polypeptide linker. ScFv play a critical role in therapy and diagnosis of human diseases, and may in fact also be developed into a potential diagnostic and/or therapeutic agent. However, the fact that current scFv antibodies have poor stability, low solubility, and affinity, seriously limits their diagnostic and clinical implication. Here we have developed four different expression vectors, and evaluated their abilities to express a soluble scFv protein. The solubility and binding activity of the purified proteins were determined using both SDS-PAGE and ELISA. Amongst the four purified proteins, the Skp co-expressed scFv showed the highest solubility, and the binding activity to antigen TLH was 3-4 fold higher than the other three purified scFv. In fact, this scFv is specific for TLH and does not cross-react with other TLH-associated proteins and could be used to detect TLH directly in real samples. These results suggest that the pACYC-Duet-skp co-expression vector might be a useful tool for the production of soluble and functional scFv antibody.

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  • Coexpression of an scFv with the periplasmic chaperones FkpA and Skp significantly improved the cytoplasmic solubility of the scFv and cell viability.

    “In another study, the coexpression of an scFv with the periplasmic chaperones FkpA and Skp significantly improved the cytoplasmic solubility of the scFv and cell viability [168,172].”

    From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [172]

    The Results (page unavailable) report improved scFv solubility with Skp co-expression. The Introduction (page unavailable) mentions FkpA and increased cell viability only when describing earlier studies; this paper did not test FkpA co-expression or report improved viability, and its solubility result is not specifically localized to the cytoplasm. (LLM, conf=0.86)

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Authors
Wang, R., Xiang, S., Feng, Y., Srinivas, S., Zhang, Y., Lin, M., & Wang, S.
Affiliations
  • Fujian Agriculture and Forestry University (CN)
  • University of Illinois Urbana-Champaign (US)
  • Waters (China) (CN)
Corresponding author
Wang, Shihua
ORCID
Venue
Frontiers in Cellular and Infection Microbiology
Publisher
Frontiers Media SA
Published
2013
Volume
3
Pages
72-72
ISSN
2235-2988 (ISSN-L)
DOI
10.3389/fcimb.2013.00072
PMC
PMC3818579
OpenAlex
W2171737066
Cited by
62
References
44
Citations by year
2026: 7 · 2025: 4 · 2024: 3 · 2023: 1 · 2022: 3 · 2021: 4 · 2020: 6 · 2019: 7 · 2018: 5 · 2017: 6 · 2016: 3 · 2015: 7 · 2014: 5 · 2012: 1
FWCI
3.88
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Frontiers in Cellular and Infection Microbiology (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Chinese Academy of Sciences
  • Xiamen University
  • University of Illinois at Urbana-Champaign
  • Program for New Century Excellent Talents in University (NCET-10)
  • Institute of Biophysics, Chinese Academy of Sciences
Topics
  • Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
Keywords
scFv, scFv antibodies, Escherichia coli expression, Skp chaperone, co-expression vector, antigen binding activity, expression vectors
MeSH terms
DNA-Binding Proteins, Electrophoresis, Polyacrylamide Gel, Enzyme-Linked Immunosorbent Assay, Escherichia coli, Genetic Engineering, Genetic Vectors, Humans, Protein Binding, Solubility, Gene Expression, Molecular Chaperones, Escherichia coli Proteins, Single-Chain Antibodies

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