VSV-Pseudovirus Lassa Virus (Josiah Strain) Glycoprotein Complex with Luciferase Reporter

Catalog Number:

M85H4

Description:

This pseudotyped virus uses recombinant vesicular stomatitis virus (rVSV) to carry glycoprotein complex (GPC) of Lassa virus (LASV) Josiah Strain (GenBank: P08669). LASV causes Lassa fever that is a severe hemorrhagic manifestations. GPC including GP1, GP2 and an unusual stable signal peptide (SSP) is responsible for receptor binding and fusion of the virion with a cell membrane. 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 >100,000-fold higher than uninfected control (cell alone as background in blue). We also evaluated the neutralizing activity of a neutralizing antibody RV32 (Catalog # M17T52) with a negative control by using this pseudotyped virus. The results showed the neutralization titer for RV32 is 1 ng/ml. See figure

Applications:

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

Lassa virus (LASV), virus caused Lassa fever.

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 500-1,000-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 1,000-2,000 reactions (100 μl diluted virus) or 2,500-5,000 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 (Vero 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 (20 μg) is free with pseudovirus purchase