Anti-CD19 CAR / NFAT (Luciferase) Reporter Jurkat Cell Line (CD19 SCFV-CD28-4-1BB-CD3ζ) | 79853

Anti-CD19 CAR / NFAT (Luciferase) Reporter Jurkat Cell Line (CD19 SCFV-CD28-4-1BB-CD3ζ) | 79853 | Gentaur US, UK & Europe Disrtribition

Category: CAR-T/Cell Line

Application: • Validate different CAR designs and constructs for their specificity, efficacy and potency before proceeding into patient-derived primary T cells.
• Predict the Mechanism of Action (MOA) of CAR.
• Intracellular co-stimulatory and activation

Background: The development of CAR T-cells is a complex process that requires multiple components in the workflow including I) screening and sequencing of mAbs that are specific to the cancer antigens; II) engineering and validation of scFv and scFv-CAR of different varieties for their specificities and activities; III) production of high titer lentivirus for CAR constructs; IV) isolation, activation and expansion of primary T cells from healthy donors or patients that exhibit a specific cellular phenotype; V) transduction of activated T cells with CAR-encoding lentivirus; V) validation of engineered CAR-T cells through FACS and functional analysis.

BPS Bioscience has developed a series of CAR-T products, including lentiviruses, reporter cell lines and fully validated functional CAR T-cells for a variety of targets such as CD19 and BCMA. In this product, anti-CD19 CAR and NFAT-luciferase reporter are co-transfected into a Jurkat cell line, where binding of CD19 to anti-CD19 scFv leads to the activation of CAR and luciferase reporter through NFAT. Anti-CD19 scFv linked to 3rd generation CAR (CD28 transmembrane and costimulatory domains, 4-1BB, and CD3ζ components) was cloned into a lentivector, and packaged using a safe, replication incompetent, VSV-G pseudotyped lentiviral packaging system, in which the gene of anti-CD19 CAR is driven by an EF-1α promotor. The anti-CD19 CAR reporter Jurkat cell line was generated by transducing the anti-CD19 CAR lentivirus into an NFAT-luciferase reporter Jurkat cell line. In these cells, the luciferase reporter is activated upon co-culture with CD19/CHO target cells (BPS Bioscience #79561), or Raji cells with endogenous CD19 expression. The anti-CD19 CAR /NFAT-luciferase reporter Jurkat cell line is a great system for primary screening of anti-CD19 CAR and predicting its mechanism of action before testing on patient-derived primary T cells. The same anti-CD19 CAR lentivirus was also used to transduce primary T cells to make primary anti-CD19-CAR T-cells, which showed IFN-γ production and cytotoxic killing of CD19+ tumor cells in co-culture experiments, indicating that there is a good correlation between the reporter activity in CAR reporter Jurkat cell line and functional activation of primary CAR T cells when co-cultured with target cells.

Description: Anti-CD19 CAR/NFAT-luciferase reporter Jurkat cell line is a double stable cell line expressing anti-CD19 CAR and NFAT-luciferase reporter. It is made from the anti-CD19 CAR lentivirus (BPS Bioscience #79851). The reporter cell line has been validated for anti CD19-CAR expression by FACS, and for luciferase reporter activation stimulated by target cells including CD19/CHO recombinant cell line can be used for primary screening and functional validation of anti-CD19 CAR construct and lentivirus before testing in primary T cells.

Anti-CD19 CAR consists of anti-CD19 scFv linked to 3rd generation CAR (Chimeric Antigen Receptor) containing CD28, 4-1BB co-stimulatory domains, and CD3ζ signaling domain.

Product Type: Cell Line

Shippement Condition: -80°C