VSV-Pseudovirus_SARS-CoV-2 Omicron BA.2.86 Strain Spike with Luciferase Reporter
Catalog Number:
W19M63
Description:
This pseudotyped virus uses recombinant vesicular stomatitis virus (rVSV) to carry spike (S) protein of SARS-CoV-2 (GenBank: MN908947) with multiple mutations initially identified in variant of Omicron BA.2.86, including: Ins16:MPLF, T19I, R21T, LPP24-26del, A27S, S50L, H69-, V70-, V127F, G142D, Y144-, F157S, R158G, N211-, L212I, V213G, L216F, H245N, A264D, I332V, G339H, K356T, S371F, S373P, S375F, T376A, R403K, D405N, R408S, K417N, N440K, V445H, G446S, N450D, L452W, N460K S477N, T478K, N481K, V483-, E484K, F486P, Q498R, N501Y, Y505H, E554K, A570V, D614G, P621S, H655Y, I670V, N679K, P681R, N764K, D796Y, S939F, Q954H, N969K, P1143L. The spike protein has a 18-aa cytoplasmic tail truncation for optimal infection. As pseudovirus infectivity of rVSV without its original G is restricted to a single round of replication, the pseudotypes can be handled using BSL-2 containment practices. Infection of cells with this pseudotyped virus carrying luciferase reporter results in high level luciferase activity. See our titration result showing that the starting 2-fold diluted pseudovirus generates signal ~50,000-fold higher than uninfected control (cell alone as background in blue). We also evaluated the neutralizing activity of one own S2-specific antibody RV6, one commercial antibody LY-CoV1404 (bebtelovimab) and one published antibody COVA1-16 (RBD-specific) by using this pseudotyped virus. The results showed that bebtelovimab showed the abolished neutralizing activity against this variant, and COVA1-16 with significant reduced activity, compared with neutralization against WT strain (Cat # P14N41). The neutralization titer for S2-specific antibody RV6 is 80 ng/ml. See figure
Applications:
- Anti-SARS-CoV-2 neutralizing antibody screening at high throughput
- Anti-SARS-CoV-2 drug screening at high throughput
- COVID-19 vaccine efficacy evaluation at high throughput
- SARS-CoV-2 pseudovirus transduction of target cells for viral entry, receptor recognition, cellular tropism and functional studies, such as ADCC analysis
Biosafety Level:
Handle it in biosafety cabinet in BSL-2. Contacted tips & tubes should be decontaminated by 10% disinfecting bleach
Organism:
SARS-Related Coronavirus 2 (virus caused COVID-19), Omicron BA.2.86 variant.
Storage and shipping condition:
Shipping with dry ice. Require -80°C storage. Multiple freeze/Thaw cycles will reduce its sensitivity. Recommend only 1 cycle. Aliquot after the first thaw.
Size:
200 μl with recommended 20-fold dilution. We recommend to use it at a dilution fold where the signal of pseudovirus infection is 1,000-fold higher than uninfected control (cell alone as background), although 100-1,000-fold high is acceptable. Can be used for 40 reactions (100 μl diluted virus) or 100 reactions if using reduced amount of pseudovirus (40 μl diluted virus).
Use protocol:
The complete protocol is shipped with the product.
Briefly, incubate 100 μl diluted virus with 100 μl your sample in each well of 96-well plate with at least one duplicate for 30-60 min at 37°C. Then add 100 μl target cells (Vero E6 or others expressing ACE2 receptor). Read luciferase signal next day. For protocol using reduced amount, incubate 40 μl diluted virus with 10 μl your sample in each well of 96-well plate for 30-60 min at 37°C. Then add 25 μl target cells. Add 150ul fresh media next day and read signal on Day 3.
Note:
- Pseudoviruses are intended for Research Use Only
- A positive control mAb is included with pseudovirus purchase
Reference:
