The improvement of anti-HER2 scFv soluble expression in Escherichia coli

Authors

  • Farzaneh Farshdari Islamic Azad University, Pharmaceutical Sciences Branch, Faculty of Advanced Sciences & Technology, Department of Biotechnology https://orcid.org/0000-0002-4214-8252
  • Maryam Ahmadzadeh Shahid Beheshti University of Medical Sciences, School of Pharmacy, Department of Pharmaceutical Biotechnology, Student Research Committee
  • Leila Nematollahi Pasteur Institute of Iran, Biotechnology Research center
  • Elham Mohit Shahid Beheshti University of Medical Sciences, School of Pharmacy, Department of Pharmaceutical Biotechnology

DOI:

https://doi.org/10.1590/s2175-97902019000317861

Keywords:

Breast cancer, HER2, scFv, Expression, Solubility

Abstract

The relationship between the expression of HER2 and malignity of breast tumors has led to the generation of antibodies targeting HER2+ tumors. In addition, the expression of scFvs, as the smallest antigen-binding region of antibody containing two disulfide bonds in Escherichia coli often results in accumulating non-functional protein in the cytoplasm. A redox-modified strain of E. coli such as Origami (DE3) may facilitate the formation of proper disulfide bond in cytoplasm. The present study aimed to optimize the expression of anti-HER2 scFv in Origami and evaluate the influence of induction temperature, and host strain on the solubility of the protein. To this aim, chemically synthesized anti-HER2 scFv of Trastuzumab was cloned in pET-22b (+). The results demonstrated that anti-HER2 scFv is expressed in Origami, purified by using Ni-NTA column, and detected by anti-His antibody in Western blot analysis. The highest anti-HER2 scFv expression in Origami was achieved 24 h after IPTG induction (1 mM) at 37 °C. Further, the total anti-HER2 scFv expression level was higher in BL21, compared to Origami strain. However, the ratio of soluble/insoluble forms of anti-HER2 scFv increased in Origami strain. Furthermore, higher soluble expression was achieved when the culture of recombinant Origami was conducted at lower temperature (25 °C).

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Published

2020-12-09

How to Cite

Farshdari, F. ., Ahmadzadeh, M. ., Nematollahi, L. ., & Mohit, E. . (2020). The improvement of anti-HER2 scFv soluble expression in Escherichia coli. Brazilian Journal of Pharmaceutical Sciences, 56, e17861 . https://doi.org/10.1590/s2175-97902019000317861

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