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Functional expression of a single-chain antibody fragment against human epidermal growth factor receptor 2 (HER2) in Escherichia coli

  • Biotechnology Methods
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The human epidermal growth factor receptor (HER) family plays an important role in cell growth and signaling and alteration of its function has been demonstrated in many different kinds of cancer. Receptor dimerization is necessary for the HER signal transduction pathway and tyrosine kinase activity. Recently, several monoclonal antibodies have been developed to directly interfere with ligand–HER receptor binding and receptor dimerization. A single chain variable fragment (ScFv) is a valuable alternative to an intact antibody. This report describes the production and purification of an ScFv specific for domain II of the HER2 receptor in Escherichia coli BL21 (DE3) cytoplasm. The majority of expressed of anti-her2his-ScFv protein was produced as inclusion bodies. A Ni-NTA affinity column was used to purify the anti-her2his-ScFv protein. The molecular weight of anti-her2his-ScFv protein was estimated to be approximately 27 kDa, as confirmed by SDS-PAGE and Western blotting assay. The anti-her2his-ScFv showed near 95 % purity and reached a yield of approximately 29 mg/l in flask fermentation. The purified anti-her2his-ScFv showed its biological activity by binding to HER2 receptor on the surface of BT-474 cells. This ScFv may be a potential pharmaceutical candidate for targeting tumour cells overexpressing HER2 receptor.

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Acknowledgments

We thank Mrs. Fatemeh Moazen for her excellent technical assistance. This work was financially supported by the Isfahan University of Medical Sciences (Grant number 189113).

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Authors and Affiliations

  1. Department of Pharmaceutical Biotechnology, Isfahan Pharmaceutical Research Center, Faculty of Pharmacy, Isfahan University of Medical Sciences, Hezar Jarib Avenue, Isfahan, Iran

    Vajihe Akbari, Hamid Mir Mohammad Sadeghi, Abbas Jafrian-Dehkordi & Daryoush Abedi

  2. Department of Chemical Engineering, University of Waterloo, 200 University Avenue, Waterloo, ON, N2L 3G1, Canada

    C. Perry Chou

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  1. Vajihe Akbari
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  2. Hamid Mir Mohammad Sadeghi
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  3. Abbas Jafrian-Dehkordi
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Correspondence to Daryoush Abedi or C. Perry Chou.

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Akbari, V., Mir Mohammad Sadeghi, H., Jafrian-Dehkordi, A. et al. Functional expression of a single-chain antibody fragment against human epidermal growth factor receptor 2 (HER2) in Escherichia coli . J Ind Microbiol Biotechnol 41, 947–956 (2014). https://doi.org/10.1007/s10295-014-1437-0

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