This pseudotyped virus uses recombinant vesicular stomatitis virus (rVSV) to carry the S protein of SARS-CoV-2 (GenBank: MN908947) with multiple mutations initially identified in variant of Omicron CH.1.1. The S has 18-aa cytoplasmic tail truncation for optimal infection.
- 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
Description: 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 over 10,000-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 all of these three antibodies showed abolished neutralizing activity against this variant.
Size: 200 μl with recommended 30-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 60 reactions (100 μl diluted virus) or 150 reactions if using reduced amount of pseudovirus (40 μl diluted virus).
Use protocol: Incubate 100 μl diluted pseudovirus with 100 μl of your sample in each well of 96-well plate with at least one duplicate for 30 to 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 to 60 min at 37 °C. Then add 25 μl target cells. Add 150 μl fresh media next day and read signal on Day 3. To be noted for neutralizing titer (such as EC50) calculation, more duplication leads to more accurate reading.
Packaging: Shipping with dry ice. Require −80 °C storage. Multiple freeze/Thaw cycles will reduce its sensitivity. Recommend only one cycle. Aliquot after the first thaw.
Caution: Handle it in biosafety cabinet in BSL-2. Contacted tips and tubes should be decontaminated by 10% disinfecting bleach. Pseudoviruses are intended for research use only.
