VSV Pseudovirus_Bombali Ebola GP with Luciferase Reporter

Catalog Number:

T42Y99

Description:

This pseudotyped virus uses recombinant vesicular stomatitis virus (rVSV) to carry GP protein of Bombali Ebolavirus (GenBank: YP_009513277.1). Bombali virus (BOMV) is a new ebolavirus and reported in 2018. 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 ~ 20,000-fold higher than uninfected control (cell alone as background in blue). This pseudovirus has been tested to neutralize against a broad neutralizing antibody RV27 and a published cross-reactive antibody FVM04 with a lesser extent (Cell Rep. 2016; 15: 1514–1526). See picture.

Applications:

• Anti-Ebolavirus neutralizing antibody screening at high throughput
• Anti-Ebolavirus drug screening at high throughput
• Ebolavirus vaccine efficacy evaluation at high throughput
• Ebolavirus 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:

Filovirus family: Ebolavirus Bombali virus (BOMV)

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 50-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 100 reactions (100 μl diluted virus) or 250 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). 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