Optimization of the Protocol for the Isolation and Refolding of the Extracellular Domain of HER2 Expressed in Escherichia coli

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Abstract


Receptor 2 of the human epidermal growth factor (HER2/neu, c-erbB2) is a 185 kDa proto-oncogene protein characterized by an overexpression in some oncological diseases, including 30% of mammary glands cancers, as well as tumors in the ovary, stomach and other organs of the human body. Since HER2- tumor status testing is the essential part of a successful cancer treatment, the expression and purification of substantial amounts of the extracellular domain (ECD) of HER2 is an important task. The production of ECD HER2 in Escherichia coli has several advantages over the use of eukaryotic expression systems, but the bulk of the recombinant product in bacteria accumulates as insoluble protein inclusion bodies. In this study, we obtained ECD HER2 in Escherichia coli as insoluble inclusion bodies and elaborated a simple, efficient, and fast protocol for the solubilization, refolding, and isolation of the protein in soluble form.


V. V. Dolgikh

Department of Microbiology, Virology and Immunology, First Pavlov State Medical University of Saint Petersburg

Author for correspondence.
Email: vtetzv@yahoo.com

Russian Federation

I. V. Senderskiy

Department of Microbiology, Virology and Immunology, First Pavlov State Medical University of Saint Petersburg

Email: vtetzv@yahoo.com

Russian Federation

G. V. Tetz

Department of Microbiology, Virology and Immunology, First Pavlov State Medical University of Saint Petersburg

Email: vtetzv@yahoo.com

Russian Federation

V. V. Tetz

Department of Microbiology, Virology and Immunology, First Pavlov State Medical University of Saint Petersburg

Email: vtetzv@yahoo.com

Russian Federation

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Copyright (c) 2014 Dolgikh V.V., Senderskiy I.V., Tetz G.V., Tetz V.V.

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