Method for producing Chinese hamster ovary cell strain, being a producer of recombinant SARS-CoV-2 virus protein RBD, Chinese hamster ovary cell strain, producer of recombinant SARS-CoV-2 protein RBD, method for producing recombinant SARS-CoV-2 virus protein RBD, test system for enzyme immunoassay of human serum or plasma, and use thereof Russian patent published in 2020 - IPC C07K14/165 C12N15/50 C07K1/36 C07K1/16 C07K16/10 

Abstract RU 2723008 C1

FIELD: biotechnology.

SUBSTANCE: The invention relates to the field of biotechnology. Created are a genetic construct for the expression of recombinant SARS-CoV-2 virus protein RBD, a strain of Chinese hamster ovary cells CHO-S-RBD that produces a recombinant protein - a receptor-binding domain (RBD) of SARS-CoV-2 virus, which can be used for diagnostic purposes. Described is a method for producing a strain of Chinese hamster ovary cells CHO-S-RBD being a producer of recombinant SARS-CoV-2 virus protein RBD, containing a genetic construct comprising SEQ ID NO: 1; introducing the specified genetic constructs into cells by lipofection; selecting cells on Hygromycin B antibiotic. Created is a strain of Chinese hamster ovary cells CHO-S-RBD, a producer of recombinant SARS-CoV-2 virus protein RBD. Developed is a method for producing recombinant SARS-CoV-2 virus protein RBD, comprising: culturing a strain of Chinese hamster ovary cells CHO-S-RBD; purifying chromatographically the recombinant SARS-CoV-2 virus protein RBD from culture medium of the Chinese hamster ovary strain CHO-S-RBD; confirming the production of recombinant SARS-CoV-2 protein RBD. Created is a recombinant SARS-CoV-2 virus protein RBD to detect antibodies to SARS-CoV-2. Created is a test system for enzyme immunoassay of human serum or plasma. Developed is a method for analyzing serum or plasma of COVID-19 convalescents to sample the most promising ones for treating patients infected with SARS-CoV-2. Developed is a method for analyzing human serum or plasma to determine the titer of antibodies to SARS-CoV-2 virus. The invention reduces time and simplifies the analysis of serum or plasma of COVID-19 convalescents to sample the most promising ones for treating SARS-CoV-2 infected patients. The invention also provides for high and stable expression of recombinant SARS-CoV-2 virus protein RBD.

EFFECT: invention provides a rapid and efficient test system for determining the most promising serum or plasma samples of COVID-19 convalescents for treating patients infected with SARS-CoV-2.

10 cl, 4 dwg, 6 tbl, 8 ex

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RU 2 723 008 C1

Authors

Shchebliakov Dmitrii Viktorovich

Esmagambetov Ilias Bulatovich

Logunov Denis Iurevich

Derkaev Artem Alekseevich

Simakin Pavel Viacheslavovich

Izhaeva Fatima Magometovna

Dzharullaeva Alina Shakhmirovna

Zubkova Olga Vadimovna

Ozharovskaia Tatiana Andreevna

Dolzhikova Inna Vadimovna

Tukhvatulin Amir Ildarovich

Tukhvatulina Natalia Mikhailovna

Favorskaia Irina Alekseevna

Naroditskii Boris Savelevich

Gintsburg Aleksandr Leonidovich

Dates

2020-06-08Published

2020-05-19Filed