Preparation method of cell for transmembrane expression of 2019-nCoV antigen, cell and application
The invention provides a preparation method, of a cell for transmembrane expression of a 2019-nCoV antigen, a cell and an application, and belongs to the technical field of gene engineering. The method comprises the following steps: integrating a sequence into a PB-713 plasmid through Bsu36I and BmgBI double restriction enzyme cutting sites, so as to obtain a PB-S-RBD-NGFR plasmid; electrically transforming the PB-S-RBD-NGFR plasmid into K562 cells, performing puromycin screening, and when the proportion of living cells is continuously larger than 80%, obtaining cells for transmembrane expression of the 2019-nCoV antigen. According to the invention, the receptor binding domain of a 2019-nCoV spike glycoprotein is independently subjected to transmembrane cell expression, so that the risk ofantibody dependence enhancement caused by other S protein epitopes is avoided, and then the receptor binding domain of the 2019-nCoV spike glycoprotein is fused with a transmembrane protein domain; and a better neutralizing titer is induced in an animal body..
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Patent| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Europäisches Patentamt - (2021) vom: 22. Jan. Zur Gesamtaufnahme - year:2021 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
JIAO SHUNCHANG [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| Anmerkungen: |
Source: www.epo.org (no modifications made), First posted: 2021-01-22, Last update posted on www.tib.eu: 2022-04-10, Last updated: 2023-02-09 |
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| Patentnummer: |
CN112251413 |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
EPA011330945 |
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| 520 | |a The invention provides a preparation method, of a cell for transmembrane expression of a 2019-nCoV antigen, a cell and an application, and belongs to the technical field of gene engineering. The method comprises the following steps: integrating a sequence into a PB-713 plasmid through Bsu36I and BmgBI double restriction enzyme cutting sites, so as to obtain a PB-S-RBD-NGFR plasmid; electrically transforming the PB-S-RBD-NGFR plasmid into K562 cells, performing puromycin screening, and when the proportion of living cells is continuously larger than 80%, obtaining cells for transmembrane expression of the 2019-nCoV antigen. According to the invention, the receptor binding domain of a 2019-nCoV spike glycoprotein is independently subjected to transmembrane cell expression, so that the risk ofantibody dependence enhancement caused by other S protein epitopes is avoided, and then the receptor binding domain of the 2019-nCoV spike glycoprotein is fused with a transmembrane protein domain; and a better neutralizing titer is induced in an animal body. | ||
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