Producing recombinant SARS-CoV-2 spike protein in a pre-fusion state
OwnerYang Xiang, Jun Liinventors
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Abstract
Disclosed is producing recombinant SARS-CoV-2 spike protein in a pre-fusion state, using furin knock out or knockdown mammalian cells (such as HEK293, CHO or other mammalian cells). The pre-fusion state SARS-CoV-2 spike protein can be used as an antigen to generate antibodies/binding molecules for use in SARS-CoV-2 detection assays or in diagnosis of active or prior infection with SARS-CoV-2; as a therapeutic to interfere with SARS-CoV-2 cellular binding; to generate antibodies/binding molecules to SARS-CoV-2 for use in therapy; or, as a vaccine for generating immunity to SARS-CoV-2; or for prophylactic or therapeutic use against related coronaviruses.
Claims10
A method of generating recombinant SARS-CoV-2 spike protein in a pre-fusion state, comprising:
- generating genetically modified host cells which lack the ability to express furin but which express SARS-CoV-2 spike protein using a CRISPR Cas9 transfection protocol having sgRNA scaffolds:
- GATGCGCACAGCCCACGTGT (SEQ ID NO:19);
- and
- ACAGTGTGGCACGGAAGCAT (SEQ ID NO:20);
- and
- expressing the SARS-CoV-2 spike protein in said host cells.
Dependencies are inferred from the claim text.