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Applications: Ideal as a positive control for anti-CD20 CAR evaluation in T cells; useful for transduction optimization.
Description: The anti-CD20 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 variable fragment) of anti-CD20 (clone Leu-16) linked to a 2nd generation CAR (Chimeric Antigen Receptor) containing CD8 hinge and transmembrane domains, and the 4-1BB and CD3ζ signaling domains .
Formulation: The lentiviruses were produced from HEK293T cells, concentrated, and resuspended in DMEM.Recommended CD4+CD8+ T Cell Medium: StemSpan SFEM (Stemcell Technologies, #09650) supplemented with 10% heat-inactivated FBS (Life Technologies, #10082147), 1% Penicillin/Streptomycin (Hyclone, #SV30010.01), plus 10 ng/ml IL-2 (BPS Bioscience, #90184).
Storage Stability: Lentiviruses are shipped with dry ice. For long term storage, it is recommended to store the virus at -80°C.
Supplied As: 50 µl of anti-CD20 CAR at a titer ≥108 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Warnings: Avoid repeated freeze/thaw cycles. Titers can drop significantly with each freeze/thaw cycle.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.
Biosafety Level: BSL-2
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