FIELD: biotechnology.
SUBSTANCE: invention relates to protein engineering, biotechnology and medicine. Described is a recombinant protein presented on SEQ ID NO: 2, having the ability to effectively bind to the receptor-binding domain of the S-protein SARS-CoV-2. Disclosed is a recombinant plasmid pET39b-LCB1-ABD-Ba, which provides synthesis of recombinant protein in Escherichia coli cells, obtained by cloning a DNA fragment presented on SEQ ID NO: 1, into the plasmid vector pET39b at the NdeI/XhoI restriction sites. Disclosed is a recombinant strain Escherichia coli BL21(DE3)/pET39b-LCB1-ABD-Ba, containing recombinant plasmid pET39b-LCB1-ABD-Ba according to cl. 2 – producer of recombinant protein. Described is a method of producing recombinant protein from biomass of recombinant strain Escherichia coli BL21(DE3)/pET39b-LCB1-ABD-Ba, at which cell destruction by disintegration is carried out, cell debris is removed, protein is recovered by means of two-step chromatography, which includes a step of protein denaturation/renaturation on metal chelate affinity sorbent and purification by means of anion-exchange chromatography. Invention can find application in biotechnology and medicine when producing drugs against COVID-19.
EFFECT: invention enables to obtain a high level of induced synthesis – 40% of total cell protein and stable production, high specific antiviral activity, prolonged circulation time and antiviral effect in blood flow in comparison with Targevir1 analogue.
4 cl, 8 dwg, 1 tbl, 9 ex
| Title | Year | Author | Number |
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| RECOMBINANT SARS-CoV-2 S PROTEIN RBD-BINDING PROTEIN | 2022 |
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