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Application: Screening for neutralizing antibodies for the Omicron variant of SARS-CoV-2 Spike in ACE2-HEK293 cells
Description: The pandemic coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). As the first step of the viral replication, the virus attaches to the host cell surface before entering the cell. The viral Spike protein recognizes and attaches to the Angiotensin-Converting Enzyme 2 (ACE2) receptor found on the surface of type I and II pneumocytes, endothelial cells, and ciliated bronchial epithelial cells. Drugs targeting the interaction between the Spike protein of SARS-CoV-2 and ACE2 may offer protection against the viral infection. A variant called B.1.1.529 (also known as the Omicron Variant) was identified in South Africa in November of 2021. This variant has a large number of mutations that allow the virus to spread more easily and quickly than other variants.The Spike (B.1.1.529 Variant) (SARS-CoV-2) Pseudotyped Lentiviruses were produced with SARS-CoV-2 B.1.1.529 Variant Spike (Genbank Accession #QHD43416.1 with B.1.1.529 mutations; see below for details) as the envelope glycoproteins instead of the commonly used VSV-G. These pseudovirions contain the eGFP gene driven by a CMV promoter, therefore, the spike-mediated cell entry can be determined via eGFP fluorescence. The Spike (B.1.1.529 Variant) (SARS-CoV-2) pseudotyped lentivirus can be used to measure the activity of neutralizing antibody against SARS-CoV-2 B.1.1.529 variant in a Biosafety Level 2 facility.The Spike Omicron pseudovirus has been validated for use with target cells ACE2-HEK293 (which overexpress ACE2; BPS Bioscience, #79951).Spike Mutations in B.1.1.529 Variant:A67V, Δ69-70, T95I, G142D, Δ143-145, Δ211, L212I, ins214EPE, G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, L981F
Formulation: The lentiviruses were produced from HEK293T cells. Supplied in medium containing 90% DMEM + 10% FBS.
Storage Stability: Lentiviruses are shipped with dry ice. For long term storage, it is recommended to store the virus at -80°C.
Supplied As: The titer will vary with each lot; the exact value is provided with each shipment.
Warnings: Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.Biosafety: None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.
Biosafety Level: BSL-2