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Anti-CD19 CAR negative control/ NFAT (Luciferase) Reporter Jurkat Cell Line (CD19 SCFV-CD28 transmembrane motif)

SKU : BHC18200222

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BPS Bioscience

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Mfr.No.	79854
Description	Anti-CD19 CAR negative control/NFAT-luciferase reporter Jurkat cell line is a double stable cell line expressing anti-CD19 CAR negative control and NFAT-luciferase reporter. The anti-CD19 CAR negative control consists of anti-CD19 scFv linked to the CD28 transmembrane motif without any intracellular signaling domains. The reporter cell line has been validated for anti- CD19 expression by FACS, while the stimulation by target cells including CD19/CHO recombinant cell line has not activated the luciferase reporter gene in this cell line. The cell line can be used for the negative control of anti-CD19 CAR/Jurkat-NFAT cell line (Biohippo, #79853). 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 steps 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; VI) 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 negative control and NFAT-luciferase reporter are co-transfected into a Jurkat cell line, where the anti-CD19 scFv binds to CD19, however, it does not induce the activation of CAR and luciferase reporter through NFAT as the intracellular activation motifs are missing. Anti-CD19 scFv linked to the CD28 transmembrane region was cloned into a lentivector, and packaged using a safe, replication incompetent, and VSV-G pseudotyped lentiviral packaging system, in which the gene of anti-CD19 CAR negative control is driven by an EF-1alpha promotor. Anti-CD19 CAR negative control Jurkat/NFAT reporter cell line was generated by the transduction of anti-CD19 CAR negative control lentivirus into an NFAT-luciferase reporter Jurkat cell line. In these cells, the luciferase reporter should not be activated upon co-culture with CD19/CHO target cells (Biohippo #79561).Anti-CD19 CAR/NFAT-luciferase reporter Jurkat cell line (Biohippo, #79853) 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 (Biohippo, #79851) 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. Intracellular co-stimulatory and activation domain comparison.
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.

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SKU	Supplier No.	Size	Your price	Quantity
BHC18200222	79854	2 vials	$1815		Add to Cart

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