Spike whole mutant SARS-CoV-2(2019nCoV) B.1.1.7 lineage Pseudotyped virus production and Pseudovirus Based Neutralization Assay
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Spike mutant variant of SARS-COV-2 (2019nCOV) B.1.1.7 lineage spreaded in UK
The world is in midst of the COVID-19 pandemic. Recently a novel SARS-COV-2 (2019nCOV) lineage, the B.1.1.7 lineage, with serials of site mutation, shows stronger infection ability in the UK. The SARS-COV-2 B.1.1.7 lineage carries a larger than a usual number of coronavirus genetic changes.
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.
In SARS-COV-2 B.1.1.7 lineage, most mutations and deletions occur in the coronavirus spike protein. These include spike position 501, one of the key contact residues in the receptor-binding domain (RBD), and experimental data suggest mutation N501Y can increase ACE2 receptor affinity (Starr et al. 2020) and P681H, one of 4 residues comprising the insertion that creates a furin cleavage site between S1 and S2 in spike. The S1/S2 furin cleavage site of SARS-CoV-2 is not found in closely related coronaviruses and has been shown to promote entry into respiratory epithelial cells and transmission in animal models (Hoffmann, Kleine-Weber, and Pöhlmann 2020; Peacock et al. 2020; Zhu et al. 2020). N501Y has been associated with increased infectivity and virulence in a mouse model (Gu et al. 2020). Both N501Y and P681H have been observed independently but not to our knowledge in combination before now.
Also present is the deletion of two amino acids at sites 69-70 in spike - this mutation is one of a number of recurrent deletions observed in the N terminal domain of Spike (McCarthy et al. 2020; Kemp et al. 2020) and has been seen in multiple lineages linked to several RBD mutations. For example, it arose in the mink-associated outbreak in Denmark on the background of the Y453F RBD mutation, and in humans in association with the N439K RBD mutation, accounting for its relatively high frequency in the global genome data (~3000 sequences).
GeneMedi pseudotype virus (pseudovirus) of SARS-COV-2 (2019nCOV) B.1.1.7 lineage
The outbreak of COVID-19, caused by SARS-CoV-2 (2019-nCoV), has been a global public health threat and caught the worldwide concern. GeneMedi has developed SARS-CoV-2 wildtype and mutation variants pseudovirus production system, from which the SARS-CoV-2 wildtype and mutation variants pseudotyped virus can be handled in biosafety level 2 (BSL-2).
GeneMedi develop COVID-19 related SARS-CoV-2 (2019-nCoV)Pseudotyped virus of SARS-CoV-2 Spike Mutation Variants including D614G,S943P,V367F, G476S, V483A, H49Y, Q239K, A831V, P1263L,D839Y/N/E:D839Y,D839N,D839E,P2-mutated Spike protein trimer variant (P2-mutant, S1/S2 cleavage site(furin cleavage sequence)-mutant, trimerization modified), Spike(S1+S2) - B.1.1.7 lineage, Spike(S1+S2)-N501Y mutation, Spike(S1+S2)-HV 69-70 Deletion mutation(ΔH69/ΔV70) and so on.
SARS-CoV-2 Pseudotyped virus with Spike D614G mutation:Increasing frequency and global distribution.
SARS-CoV-2 Pseudotyped virus with Spike S943P mutation:It is found only in Belgium
SARS-CoV-2 Pseudotyped virus with Spike V367F mutation:
SARS-CoV-2 Pseudotyped virus with Spike G476S mutation
SARS-CoV-2 Pseudotyped virus with Spike V483A mutation
SARS-CoV-2 Pseudotyped virus with Spike H49Y mutation
SARS-CoV-2 Pseudotyped virus with Spike Q239K mutation
SARS-CoV-2 Pseudotyped virus with Spike A831V mutation
SARS-CoV-2 Pseudotyped virus with Spike P1263L mutation
SARS-CoV-2 Pseudotyped virus with Spike D839Y/N/E-D839Y mutation
SARS-CoV-2 Pseudotyped virus with Spike D839Y/N/E-D839N mutation
SARS-CoV-2 Pseudotyped virus with Spike D839Y/N/E-D839E mutation
SARS-CoV-2 Pseudotyped virus with P2-mutated Spike protein trimer variant (P2-mutant, S1/S2 cleavage site (furin cleavage sequence)-mutant,trimerization modified) mutation
SARS-CoV-2 Pseudotyped virus with Spike of SARS-COV-2 B.1.1.7 lineage mutation
SARS-CoV-2 Pseudotyped virus with Spike(S1+S2) N501Y mutation mutation
SARS-CoV-2 Pseudotyped virus with Spike(S1+S2)-HV 69-70 Deletion mutation(ΔH69/ΔV70) mutation
GeneMedi’s Pseudovirus Based Neutralization Assay (PBNA) is a conventional assay method that is suitable for High-Throughout Screening (HTS) without live virus engaged. The Pseudovirus Based Neutralization Assay can be used for evaluating:
1) Neutralizing antibodies
2) Peptides blockers (peptide inhibitors)
3) Types of Vaccines3
4) Compounds targeting Spike induced cell-fusion.
1.SARS-CoV-2 Pseudotyped virus packaging and production
2.Effector cells: human ACE2 overexpression stable HEK293T cell lines
3.SARS-CoV-2(2019nCoV) Pseudotyped Virus Based Neutralization Assay service.