The Anti-mesothelin CAR lentiviruses are replication incompetent, HIV-based, VSV-G-pseudotyped lentiviral particles that are ready to infect almost all types of mammalian cells, including primary and non-dividing cells. These viruses transduce the ScFv (single-chain fragment variable) of anti-mesothelin (Clone P4) linked to a 2nd generation CAR (Chimeric Antigen Receptor) containing CD8 hinge and transmembrane domains, and the 4-1BB and CD3ζ signaling domains.
Mesothelin (MSLN) is a glycophosphatidylinositol (GPI) linked cell-surface protein that is produced as a ~70 kDa precursor protein and cleaved by Furin protease to generate the ~40 kDa mature form. MSLN is frequently overexpressed in mesothelioma, ovarian, pancreatic, and non-small cell lung cancers, while its expression in normal tissues is restricted to the mesothelial lining. MSLN is a tumor-associated antigen and has been an attractive target for targeted immunotherapy, including drug-conjugated antibodies and chimeric antigen receptor T cells (CAR-T Cells).
The lentiviruses were produced from HEK293T cells, concentrated, and resuspended in DMEM.
Biosafety
The lentiviruses are produced with a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.
Positive control for anti-mesothelin CAR evaluation in T cells; useful for transduction optimization.
Storage
Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C for up to 12 months from date of receipt. Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.
Shipping
-80°C
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Reviews of Anti-Mesothelin CAR Lentivirus (P4 ScFv-CD8-4-1BB-CD3zeta)