Lentivirus Pseudotyped Influenza A HPAI H5N1 virus HaNaM with Luciferase Reporter (clade 2.3.4.4b, 2022 strain)

Catalog Number:

S45R27

Description:

This pseudotyped virus uses lentivirus packaging to carry full length HA, NA and M of Highly Pathogenic Avian Influenza (HPAI) A virus (A/Pelecanus/Peru/VFAR-140/2022(H5N1), clade 2.3.4.4b) (GenBank: OQ565628.1). HPAI influenza H5N1 virus that is highly contagious among birds but also infect humans should be handled in BSL-4 labs. While 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 published neutralizing mAb 100F4 (Nat Commun. 2015:6:8855) and two control mAbs by using this pseudotyped virus. See figure

Applications:

• Anti-avian influenza virus neutralizing antibody screening at high throughput
• Anti-avian influenza virus drug screening at high throughput
• Avian influenza virus vaccine efficacy evaluation at high throughput
• Avian influenza 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:

Pathogenic Avian Influenza (HPAI) A virus, H5N1, clade 2.3.4.4b, 2022 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 10-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 20 reactions (100 μl diluted virus) or 50 reactions if using reduced amount of pseudovirus (40 μl diluted virus).

Use protocol:

A complete protocol is supplied with the shipping of 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 (293T or other lung cells 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 included with pseudovirus purchase