Recombinant 2019-nCoV(SARS-CoV-2) Spike S-trimer Protein (Omicron variant aka B.1.1.529 lineage) with furin cleavage site mutation & T4 fibritin trimerization motif

Cat No.: GMP-V-2019nCoV-S-trimer-004

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Package Catalog No. Price(In USD) Qty (Quantity) Sum(In USD)
1mg GMP-V-2019nCoV-S-trimer-004-1mg 5990
10mg GMP-V-2019nCoV-S-trimer-004-10mg 38790
100mg GMP-V-2019nCoV-S-trimer-004-100mg 258000
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Description

Accession Number QHD43415.1
Expression platform 2019-nCoV(SARS-CoV-2, SARS2-coronavirus)
IsotypesMamamlian (human cell)
TagC-His
Products descriptionRecombinant 2019-nCoV(SARS-CoV-2) Spike S-trimer Protein (Omicron variant aka B.1.1.529 lineage) with furin cleavage site mutation & T4 fibritin trimerization motif was expressed in mammanlian cell (human cells) expression system with 6 HIS tag at the C-terminus.
Bioactivity validation ACE2 binding;
Immunogen in Sandwich Elisa, lateral-flow tests,and other immunoassays;
PurityPurity: ≥95% (SDS-PAGE)
ApplicationSpike protein & ACE2 competition binding assay for efficacy evaluation of COVID-19 vaccines and therapeutic antibodies.
Immunogen in Elisa,lateral-flow tests,and other immunoassays;
Standard substance
The antigen can also be used in drug discovery including antibody screening and lead compound candidates assay.
predicted Molecular Mass136kDa
Formulation & ReconstitutionLyophilized from GM's Protein Stability Buffer2 (PSB2,Confidential Ingredients) or PBS (pH7.4);
For PSB2, reconstituted with 0.9% sodium chloride;
For PBS, reconstituted with ddH2O.
StorageStore at -20℃ to -80℃ under sterile conditions. Avoid repeated freeze-thaw cycles.

COVID-19 recombinant SARS-CoV2 (2019 nCoV coronavirus) antigen:
Recombinant 2019-nCoV(SARS-CoV-2) Spike S-trimer protein S6P mutant (F817P, A892P, A899P, A942P, K986P, V987P).

The target gene encoding Met1-Gln1208 (RR682G, R683S, R685S, F817P, A892P, A899P, A942P, K986P, V987P) was expressed with 6 HIS tag at the C-terminus. Proline substitutions (F817P, A892P, A899P, A942P, K986P, V987P) and S1/S2 cleavage site (furin cleavage sequence) mutation of PRRAR to PGSAS at residues 682–685 (R682G, R683S, R685S) are introduced to stabilize the trimeric prefusion state of SARS-CoV-2 S protein and abolish the furin cleavage site, respectively. T4 fibritin trimerization motif is added to C-terminal of the protein to improve the stability of spike trimer protein.

Suitable for variants of Spike protein & ACE2 competition binding assay for efficacy evaluation of COVID-19 vaccines and therapeutic antibodies.

Validated in functional ELISA, other immunoassays, the antigen can also be used in drug discovery including antibody screening and lead compound candidates assay.



Information of SARS-CoV-2 (2019nCOV) Spike protein trimer

The world is in midst of the COVID-19 pandemic caused by SARS-CoV-2 (2019nCoV) infection.The Spike protein (S-protein) of SARS-CoV-2 (2019nCoV) mediates receptor (ACE2) binding and cell entry and is the dominant target of the immune system. The highly flexible Spike protein (S-protein), with its mobile domains, transitions from closed to open conformations to expose its receptor-binding site and, subsequently, from prefusion to postfusion conformations to mediate fusion of viral and cellular membranes. SARS-CoV-2 (2019nCoV) Spike protein (S-protein) derivatives are components of vaccine candidates and diagnostic assays, as well as tools for research into the biology and immunology of SARS-CoV-2. We designed Spike mutations that allow the production of thermostable, disulfide-bonded Spike-protein trimers that are trapped in the closed, prefusion state. Structures of the disulfide-stabilized and non-disulfide-stabilized proteins reveal distinct closed and locked conformations of the Spike protein trimer.

The thermostable, disulfide-bonded Spike-protein trimers construct we make is:
1) S1/S2 cleavage site (furin cleavage sequence) mutant: Modification of the coding sequence of the multibasic S1/S2 cleavage site (furin cleavage sequence) PRRAR to PGSAS at residues 682–685 (R682G, R683S, R685S).
2) S-P6 mutant: proline substitutions: F817P, A892P, A899P, A942P, K986P, V987P.
3) Trimerization statues: add exogenous trimerization signal peptides: T4 fibritin trimerization motif and a polyhistidine tag at the C-terminus.

It has been demonstrated that the designed, thermostable, closed Spike trimer can be used in serological assays in COVID-19 test. This trimer protein has potential applications as a reagent for serology, virology and as an immunogen of COVID-19 related diagnostics and vaccine development. GeneMedi also offers pre-made gene ORF mutation plasmids of Spike trimer mutation(thermostable, disulfide-bonded Spike-protein trimers).

The pre-made gene ORF vector of Spike trimer mutation is Codon Optimized for mamamlian and can be used for mammalian expression.

GeneMedi also provides pre-made the lentivirus, adenovirus and AAV vector for the gene ORF plasmids of 2019 nCoV (SARS2 coronavirus).

Reference:
Xiong X, Qu K, Ciazynska KA, Hosmillo M, Carter AP, Ebrahimi S, Ke Z, Scheres SHW, Bergamaschi L, Grice GL, Zhang Y; CITIID-NIHR COVID-19 BioResource Collaboration, Nathan JA, Baker S, James LC, Baxendale HE, Goodfellow I, Doffinger R, Briggs JAG. A thermostable, closed SARS-CoV-2 spike protein trimer. Nat Struct Mol Biol. 2020 Oct;27(10):934-941. doi: 10.1038/s41594-020-0478-5. Epub 2020 Jul 31. PMID: 32737467; PMCID: PMC7116388.