VSV Pseudotyped HCV E1E2 with Luciferase Reporter (H77 strain)
Catalog Number:
P2R77
Description:
This pseudotyped virus uses recombinant vesicular stomatitis virus (rVSV) to carry the full length E1E2 of Hepatitis C virus (HCV) (H77 strain) (GenBank: AAB67036.1). E1 and E2 are heavily glycosylated type I transmembrane proteins that associate to form a noncovalent heterodimer – E1E2. E2 is the major target for neutralizing mAbs. As pseudovirus infectivity of rVSV without its original G is restricted to a single round of replication, and this pseudovirus is constructed only to carry full-length E1E2 of this viral strain, 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 ~100,000-fold higher than uninfected control (cell alone as background in blue). We also evaluated the neutralizing activity of one positive antibody RV36 (Cat.No. X81H49) by using this pseudotyped virus. See figure
Applications:
- Anti-Hepatitis C virus neutralizing antibody screening at high throughput
- Anti-Hepatitis C virus drug screening at high throughput
- Hepatitis C virus vaccine efficacy evaluation at high throughput
- Hepatitis C virus 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:
Hepatitis C virus (HCV), H77 strain
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 500-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 1,000 reactions (100 μl diluted virus) or 2,500 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 with at least one duplicate for 30-60 min at 37°C. Then add 100 μl target cells (Huh-7 or others expressing 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 free with pseudovirus purchase
