The BCMA Lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. The particles contain a human BCMA (NM_ 001192) driven by a CMV promoter and a G418 selection marker (Figure 1).
Figure 1: Schematic of the lenti-vector used to generate the BCMA Lentivirus
Two vials (500 µl x 2) of lentivirus at a titer ≥107 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Formulation
The lentivirus particles were produced from HEK293T cells. They are supplied in cell culture medium containing 90% DMEM + 10% FBS.
UniProt #
Q02223
Background
B-cell maturation antigen (BCMA, also known as CD269 or tumor necrosis factor receptor superfamily member 17, TNFRSF17), is a cell surface receptor of the TNF receptor superfamily that recognizes B-cell activating factor (BAFF). BCMA is preferentially expressed in mature B lymphocytes and also on Multiple Myeloma (MM) cells. BCMA is a highly attractive target antigen for immunotherapy because of its restricted expression in non-malignant tissue but almost universal expression on MM cells. CAR T cells targeting BCMA have clinical anti-MM activity, and in 2017, the FDA granted BCMA CAR T-Cell immunotherapy breakthrough designation in Multiple Myeloma.
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.
Applications
Generate a stable cell line expressing human BCMA with G418 selection and limiting dilution
Shipping Temperature
-80°C
Notes
To generate a BCMA stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of G418 (as pre-determined from a killing curve) for antibiotics selection of transduced cells. Visit: https://bpsbioscience.com/cell-line-faq for guidelines on performing a kill curve.
Biosafety 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 and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.
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