P2-mutated Spike protein trimer variant - (K986P,V987P, S1/S2 cleavage site (furin cleavage sequence)-mutant, trimerization modified) mutation SARS-CoV-2(2019nCoV) Pseudotyped virus production and Pseudovirus Based Neutralization Assay

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GeneMedi Spike P2& S1/S2 cleavage site (furin cleavage sequence) mutant SARS-CoV-2(2019nCoV) Pseudotyped virus production and Pseudovirus Based Neutralization Assay.

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 P2-mutated Spike protein trimer variant vector (P2-mutant,S1/S2 cleavage site（furin cleavage sequence）-mutant, trimerization modified)

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.
2) P2 mutant: proline substitutions at residues 986 and 987.
3) Trimerization statues: add exogenous trimerization signal peptides.

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 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).