VSV-Pseudovirus_SARS-CoV-2 D614G mutant Spike with luciferase reporter
Catalog Number:
M32H50
Description:
This pseudotyped virus uses recombinant vesicular stomatitis virus (rVSV) to carry a mutated S protein from D (Asp) to G (Gly) at site of 614 of SARS-Cov-2 (GenBank: MN908947), which has a 18-aa cytoplasmic tail truncation for optimal infection. D614G mutant was found to be prevalent from April 2020 and enhances viral transmission (Cell 182, 1-16; 2020). 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 the undiluted pseudovirus (red) generates signal 100,000-fold higher than uninfected control (cell alone as background in blue), and 10-fold higher than WT pseudovirus (green). We also evaluated the neutralizing activity of one SARS2 Positive control mAb by using this pseudotyped virus, see results.
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)
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 300-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 600 reactions (100 μl diluted virus) or 1,500 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 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:
