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 (Biohippo #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 CD3zeta signaling domain.
Mycoplasma testing: The cell line has been screened using Lonza MycoAlert Mycoplasma Detection kit (Lonza, #LT07-318) to confirm the absence of Mycoplasma species.
Host Species: human
Supplied as Each vial contains 2 x 10^6 cells in 1 ml of 10% DMSO and 90% FBS
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.
Biohippo 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 CD3zeta 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-1alpha 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 (Biohippo #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-gamma 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.
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
Host Cell
Jurkat
Shipping Temperature
-80°C (dry ice)
Note
License Disclosure: Purchase of this cell line grants you with a 10-year license to use this cell line in your immediate laboratory, for research use only. This license does not permit you to share, distribute, sell, sublicense, or otherwise make the cell line available for use to other laboratories, departments, research institutions, hospitals, universities, or biotech companies. The license does not permit the use of this cell line in humans or for therapeutic or drug use. The license does not permit modification of the cell line in any way. Inappropriate use or distribution of this cell line will result in revocation of the license and result in an immediate cease of sales and distribution of Biohippo products to your laboratory. Biohippo does not warrant the suitability of the cell line for any particular use, and does not accept any liability in connection with the handling or use of the cell line. Modifications of this cell line, transfer to another facility, or commercial use of the cells may require a separate license and additional fees; contact [email protected] for details. Publications using this cell line should reference Biohippo Inc. Warning: Avoid freeze/thaw cycles.
Storage Stability
Immediately upon receipt, store in liquid nitrogen.
Reference
1. Immune checkpoint blockade and CAR-T cell therapy in hematologic malignancies.Wang et al. J Hematol Oncol. 2019 Jun 11;12(1):59-78. 2. Chimeric antigen receptor T cell therapy for multiple myeloma. Hasegawa et al. Inflamm Regen. 2019 Jun 4;39:10-14. 3. Novel targets for the treatment of relapsing multiple myeloma. Giuliani et al. Expert Rev Hematol. 2019 Jun 3:1-16. 4. Anti-CD19 antibodies in the future management of multiple myeloma. Gavriatopoulou et al. Expert Rev Anticancer Ther. 2019 Apr;19(4):319-326.