Anti-CD19 CAR Lentivirus (CD19 ScFv-CD8-4-1BB-CD3ζ, eGFP)

Catalog #

78775

$1,350 *

Size: 50 µl

Qty

Bulk/Custom

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*US Pricing only. For international pricing, please contact your local distributor.

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Description

The anti-CD19 CAR lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to ready to transduce most mammalian cells, including primary and non-dividing cells. These viruses transduce the ScFv portion of anti-CD19 (clone FMC63) linked to 2nd generation CAR (Chimeric Antigen Receptor), containing CD8 hinge and transmembrane domains, 4-1BB and CD3ζ signaling domains. This construct also includes an IRES- enhanced green fluorescent protein (eGFP) sequence downstream of anti-CD19 CAR cassette to facilitate analysis and sorting of transduced cells (Figure 1).

Figure 1. (A) Schematic of the lenti-vector used to generate the anti-CD19 CAR lentivirus. The vector is a SIN vector, and it contains eGFP. (B) Construct diagram showing components of the anti-CD19 CAR.

Note: This product transduces the same anti-CD19 CAR construct (CD19 ScFv-CD8-4-1BB-CD3ζ) as other available anti-CD19 CAR Lentiviruses (BPS Bioscience #78600, 78601 and 78602), but differ in key aspects. This product contains an eGFP reporter/selection marker so the transduced cells can be sorted based on eGFP expression. Please see table below.

BPS Bioscience Catalog #	Self-Inactivation (SIN)	Selection Marker
78600	no	puromycin
78601	yes	no
78602	yes	puromycin
78775	yes	eGFP

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Product Data Gallery

Expression of anti-CD19 CAR and eGFP in T cells transduced with anti-CD19 CAR lentivirus.

Cytotoxicity profile of T cells transduced with anti-CD19 CAR Lentivirus against CD19/Firefly Luciferase CHO target cells.

Product Info

Storage and Usage

Citations

Supplied As

50 µl of anti-CD19 CAR at a titer ≥ 3x10⁸ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Materials Required But Not Supplied

Name	Ordering Information
PBMC, Frozen	BPS Bioscience #79059
Human Interleukin-2	BPS Bioscience #90184
EasySep™ Human CD4⁺ T Cell Isolation Kit	Stemcell Technologies #17952
EasySep™ Human CD8⁺ T Cell Isolation Kit	Stemcell Technologies #17953
Human CD3/CD28/CD2 T Cell Activator	Stemcell Technologies #10970
PE-Labeled Anti-FMC63 scFv Monoclonal Antibody	Acrobiosystems #FM3-HPY53-25tests
CD19 / Firefly Luciferase - CHO Recombinant Cell Line	BPS Bioscience #79714
Firefly Luciferase - CHO Recombinant Cell Line	BPS Bioscience #79725
ONE-Step™ Luciferase Assay System	BPS Bioscience #60690

Formulation

The lentiviruses were produced from HEK293T cells, concentrated, and resuspended in DMEM. Virus particles can be packaged in custom formulations by special request, for an additional fee.   

Background

CD19 (also known as Cluster of Differentiation 19, B-lymphocyte surface antigen B4, or CVID3) is a glycoprotein expressed at the surface of B lymphocytes through most phases of B cell maturation. It is strictly required for B cell terminal differentiation. Mutations in the CD19 gene cause severe immune-deficiency syndromes associated with impaired antibody production such as CVID3 (common variable immuno-deficiency 3). The majority of B cell malignancies express normal to high levels of CD19, making it a nearly ideal target for cancer immunotherapy. Blinatumomab, a CD19/CD3 bi-specific T cell engager (BiTE) has been approved for relapsed/refractory B precursor ALL (Acute lymphoblastic leukemia) and CD19 was the target of the first approved CAR-T cell therapy. Studies of CD19 function and expression profiles will continue to broaden our knowledge and support broader applications in cancer therapy.

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.

Growth Media

T Cell Medium: TCellM™ (BPS Bioscience #78753) supplemented with 10 ng/ml Interleukin-2 (BPS Bioscience #90184).

Media Required for the Proposed Assay

Thaw Medium 3 (BPS Bioscience #60186): F-12K Medium (Kaighn's Modification of Ham's F-12 Medium) supplemented with 10% FBS, 1% Penicillin/Streptomycin.

Applications

Positive control for anti-CD19 CAR evaluation in T cells.
Transduction optimization experiments.
Generate anti-CD19 CAR-T cells (for research use only, not for therapeutic purposes).

Shipping Temperature

-80°C

Notes

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.

License Disclosure
Visit bpsbioscience.com/license for the label license and other key information about this product.

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For all further questions, please email [email protected].

Warnings

Avoid freeze/thaw cycles

Scientific Category

Immunotherapy

Regulatory Status

Research Use Only

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Anti-CD19 CAR Lentivirus (CD19 ScFv-CD8-4-1BB-CD3ζ, eGFP)

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