Abstract
A recombinant strain for producing human gamma interferon (IFN-γ) E. coli BL 21/pET-IFN-γ was constructed to provide a high level of its expression. A method has been developed for obtaining a soluble form of recombinant IFN-γ, consisting of the processes of producing a biomass of a producer strain containing a target protein at a level of 32–37% of the total content of cellular proteins, protein isolation, and purification. The purification process included the stages of disintegration, clarification of the cell lysate, chromatographic purification, and dialysis. The developed method makes it possible to obtain up to 5 mg of the drug from 1 g of wet biomass with a purity of at least 95% and high specific (antiviral) activity.
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ACKNOWLEDGMENTS
The team of authors would like to express their gratitude to the employees of the OBTC FBUN GNTs VB Vektor of Rospotrebnadzor: Usovoi S.V., Peryshkina E.F., Bashkina E.S. for conducting experiments to determine the specific (antiviral) activity of the IFN-γ preparation.
Funding
The work was carried out within the framework of the state task, Subject GZ-39/21 Development of the technology for the preparative production and purification of recombinant proteins.
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The authors declare that they have no conflicts of interest. This article does not contain a description of the studies performed by the authors with the participation of people or animals as objects of study.
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Volosnikova, E.A., Esina, T.I., Shcherbakov, D.N. et al. The Production of Soluble Human Gamma Interferon in the Escherichia coli Expression System with a Decrease in Cultivation Temperature. Appl Biochem Microbiol 59, 132–137 (2023). https://doi.org/10.1134/S0003683823020138
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DOI: https://doi.org/10.1134/S0003683823020138