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Application: Positive control for anti-CD20 CAR evaluation in T cells.Transduction optimization experiments.Generate anti-CD20 CAR-T cells (for research use only, not for therapeutic purposes).
Background: CD20 (also known as MS4A1) is a glycosylated phosphoprotein expressed on the cell surface of B cells. CD20 is a highly attractive target antigen for immunotherapy because it is highly expressed in more than 90% of patients with B-cell lymphoma. First approved in 1997, Rituximab (Rituxan) is a chimeric monoclonal antibody targeting CD20 and has been classified by the World Health Organization as an “Essential Medicine”. Since then, additional monoclonal antibodies against CD20 have been approved or are being tested in clinical trials for the treatment of multiple sclerosis (MS), chronic lymphocytic leukemia (CLL), follicular lymphoma, diffuse large B cell lymphoma (DLBCL), rheumatoid arthritis, non-Hodgkin’s lymphoma, systemic lupus erythematosus, and myalgic encephalomyelitis (chronic fatigue syndrome). More recently, anti-CD20-CD19 bispecific CAR-T cells have been developed to address concerns over potential relapse in cancer patients.
Description: The Anti-CD20 CAR Lentivirus (Clone 3H7-CD8-41BB-CD3ζ) are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce all types of mammalian cells, including primary and non-dividing cells. These viruses transduce the ScFv portion of anti- CD20 (clone 3H7) linked to a 2nd generation CAR (Chimeric Antigen Receptor), containing the CD8 hinge and transmembrane domains, 4-1BB and CD3ζ signaling domains (Figure 1).These lentiviruses have been validated by flow cytometry (to determine the CAR expression) and in co-culture cytotoxicity assays.A. B. Figure 1: Anti-CD20 CAR Lentivirus (Clone 3H7-CD8-41BB-CD3ζ) construct diagrams.(A) Schematic of the lenti-vector used to generate the anti-CD20 CAR lentivirus. (B) Construct diagram showing components of the anti-CD20 CAR.
Formulation: The lentiviruses were produced from HEK293T cells, concentrated, and resuspended in DMEM. Virus particles can be packaged in custom formulations upon special request, for an additional fee.
Storage Stability: 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.
Supplied As: 50 µl of anti-CD20 CAR at a titer ≥ 3 x 108 TU/ml. The titer may vary with each lot; the exact value is provided with each shipment.
Target: CD20
Uniprot: P11836
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
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