Anti-CD19 CAR-T Cells

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78171

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Description

The anti-CD19 CAR-T cells are produced by high-titer lentiviral transduction of human primary CD4+CD8+ T cells using the anti-CD19 CAR Lentivirus (CD19 ScFv-CD8-4-1BB-CD3ζ; SIN Vector, BPS Bioscience #78601). These ready-to use CAR-T cells express an anti-CD19 CAR consisting of the ScFv (Single chain fragment variable) of anti-CD19 (clone FMC63) linked to a 2^nd generation CAR (Chimeric Antigen Receptor) containing CD8 hinge and transmembrane domains, and the 4-1BB and CD3ζ signaling domains (Figure 1).

These CAR-T cells have been validated using flow cytometry (to determine the CAR expression) and co-culture cytotoxicity assays.

Figure 1: Construct diagram showing components of the anti-CD19 CAR expressed in anti-CD19 CAR-T cells.

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Product Data Gallery

Expression of anti-CD19 CAR in anti-CD19 CAR-T cells

Luciferase-based cytotoxicity assay using CD19-Luciferase CHO cells as the target cells

Luciferase-based cytotoxicity assay using Firefly Luciferase-Raji Recombinant Cell Line as the target cells

Product Info

Storage and Usage

Citations

Supplied As

Each vial contains 2 x 10⁶ cells in 1 ml of CryoStor® CS10 (Stemcell Technologies)

Materials Required But Not Supplied

Name	Ordering Information
Human Interleukin-2	BPS Bioscience #90184
Human CD3/CD28/CD2 T Cell Activator	Stemcell technologies #10970
CD19/Firefly Luciferase CHO Cell Line	BPS Bioscience #79714
Firefly Luciferase CHO Cell Line	BPS Bioscience #79725
Firefly Luciferase Raji Cell Line	BPS Bioscience #78622
Firefly Luciferase K562 Cell Line	BPS Bioscience #78621
Untransduced T Cells (Negative Control for CAR-T cells)	BPS Bioscience #78170
ONE-Step™ Luciferase Assay System	BPS Bioscience #60690
Luminometer

Recommended anti-CD20 CAR-T Cell Medium: TCellM™ (BPS Bioscience #78753) supplemented with 10 ng/ml Interleukin-2 (BPS Bioscience #90184).

Mycoplasma Testing

The cells have been screened to confirm the absence of Mycoplasma species.

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, which is 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). In addition, CD19 was the target of the first approved CAR-T cell therapy.

Storage/Stability

Cells are shipped in dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage.

Applications

Use as positive control for anti-CD19 CAR-T cells
Screen modulators of anti-CD19 CAR-T cytotoxicity
Design and optimize co-culture cytotoxicity assays

Shipping Temperature

-80°C

Notes

Biosafety
The anti-CD19 CAR-T cells are produced with the third generation 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 and are not present in the lentivirus particle.

Troubleshooting Guide
Visit Cell Line FAQs for more information
For all further questions, please email [email protected].

Warnings

Avoid freeze/thaw cycles

Donors have been screened and determined negative for:

Hepatitis B (anti-HBc EIA, HBsAg EIA)
Hepatitis C (anti-HCV EIA)
Human Immunodeficiency Virus (HIV-1/HIV-2 plus O)
Human T-Lymphotropic Virus (HTLV-I/II)
HIV-1/HCV/HBV
West Nile Virus
Trypanasoma cruzi

Note: Testing cannot guarantee that any sample is completely virus-free. These cells should be treated as potentially infectious and appropriate biological safety level 2 precautions should be used.

Scientific Category

CAR-T

Regulatory Status

Research Use Only

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Anti-CD19 CAR-T Cells

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