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 BA. The S has 18-aa cytoplasmic tail truncation for optimal infection.

• 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

Description: Infection of cells with this pseudotyped virus carrying luciferase reporter results in high level luciferase activity. Titration result shows that the starting 2-fold diluted pseudovirus generates signal approximately 50,000-fold higher than uninfected control (cell alone as background in blue). It was also evaluated the neutralizing activity of one 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. The neutralization titer for S2-specific antibody RV6 is 80 ng/ml.

Use protocol: Use a small aliquot to test your own systems and use our data as reference. Briefly, 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.