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The anti-CD20 chimeric antigen receptor (CAR) Jurkat cell line is a stable cell line derived from anti-CD20 CAR lentivirus transduction. This transduced CAR lentiviral vector was constructed to express scFv of an anti-CD20 antibody linked to the TM28-CD28-CD3ζ signaling domains. And this recombinant cell product can be used to target human CD20 and treat human CD22 and Acute lymphoblastic leukemia (ALL).
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Specifications
Species
Human
Host Cell Type
T lymphocyte
Overexpressed Gene
CD20
Culture Condition
Suspension
Growth Pattern
Exponential growth phase
Formulation
Containing ≥ 1 X 10*6 / vial frozen cells
Cell Purity
>95%
Cell Viability
>90%
Mycoplasma Testing
The cell line has been screened using the luciferase based mycoplasma detection kit to confirm the absence of mycoplasma species.
Shipping
Dry ice
Storage
Frozen cells should be stored in a liquid nitrogen tank (-150°C~-190°C). Once reconstituted, the cells may be used for up to five days if properly stored at 2°C - 8°C in the buffer provided.
Warnings
Avoid multiple freeze/thaw cycles
Research Use Only
Our recombinant Jurkat cell are for research use only, not for diagnostic or therapeutic use.
Quality Control
Cultures are screened for the presence of bacteries, yeast, fungi and mycoplasma (DNA amplification). Growth media are also certified based on U.S. Public Health Service Guidelines.
Tumorgenicity
Positive (In vitro/vivo transformation assay)
Oncogenicity
Positive (In vitro soft agarose assay and life-time studies )
Sterility Testing
Creative Biolabs provides sterility testing in accordance with USP and EP regulations. All of our sterility testing is performed in an isolator or clean room environments. The cell line has been screened using the membrane filtration testing methods to confirm the absence of aerobic, anaerobic and fungi microorganisms.
Identity Testing
Identity testing is required for newly established cell lines. Isoenzyme analysis is used to confirm the identity of the species of a cell line. Alternative methods for identity testing include DNA fingerprinting, STR analysis and karyology.
Virological Safety Testing
A broad range of viruses is susceptible to affecting human cell lines. We can provide in vivo/vitro virus saftey assays by utilizing various animal systems. These viruses include: adventitious viruses, bovine viruses, human and simian viruses, porcine viruses, retrovirus and rodent viruses.
Genetic Stability Testing
We perform cell genetic stability studies under ICH guidelines. We can provide guidance on the appropriate testing program upon your requirements.
PROTOCOL
1. Thaw CAR-T cell samples quickly in a 37°C water bath until all visible ice has melted. Thaw time for a 1 ml sample in a cryovial is 2-3 minutes. Cryovials should be cool to the touch when removed from the water bath. 2. Dilute cell/medium mixture immediately with CAR-T cell culture medium. This can be performed in a single step. The dilution medium should be between 20-37°C. A dilution ratio of 1:10 (sample:medium) or greater is recommended. 3. Plate cells appropriately according to the experimental conditions of assays. 4. Culture the control lentivirus CAR-T cells or use immediately Note: T cells that are used immediately after thawing have the highest level of viability.
Application
1. Compound screening; 2. Antibody screening; 3. Co-stimulatory and activation domain comparison; 4. Personalized medicine and donor variations for CAR-T screening; 5. Checkpoint inhibitors; 6. Safety switches and regulators of CAR-T functions; 7. Pre-clinical in vivo models; 8. Treg and T memory cells in CAR-T setting; 9. CAR-T signaling, tumor microenvironment; 10. Proof of concept studies for clinical trials;
Images
Figure 1 Structure
CAR Design
Target
CD20
Target Species
Human
CAR Construction
scFv-TM28-CD28-CD3ζ
CAR Signaling Cassetes
CD28 CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide costimulatory signals required for T cell activation and survival. CD28 is the receptor for CD80 (B7.1) and CD86 (B7.2) proteins which are expressed on antigen-presenting cells (APC). CD28 modulates the primary TCR/CD3ζ signal in a different fashion than the late costimulatory elements OX40 and 4-1BB. CD28 enhances the expression of downstream regulators that impact on T-cell proliferation, death, differentiation, and effector functions. CAR+ T cells containing the CD28 endodomain showed strikingly enhanced sustained T cell activation, growth, survival. And CD28 results in a brightly expressed, stable receptor as the transmembrane domain. Including CD28 costimulatory domains in CARs led to enhanced anti-malignancy efficacy. CD3ζ CD3ζ, also known as T-cell receptor zeta, which together with T-cell receptor and CD3γ, δ , ε chain, forms the TCR-CD3 complex. ζ was expressed independently from the complex. The zeta chain plays an important role in coupling antigen recognition to several intracellular signal-transduction pathways. CD3-zeta, which contains 3 ITAMs, is the most commonly used endodomain component of CARs. It transmits an activation signal to the T cell after antigen is bound. CD3-zeta may not provide a fully competent activation signal and additional co-stimulatory signaling is needed. For example, chimeric CD28 and OX40 can be used with CD3-zeta to transmit a proliferative/survival signal, or all three can be used together.
Cannot find the desired product? Don't worry, just try our online CAR and CAR cell customizing system, which offers full options to meet all unique needs, including but not limited to conventional or unconventional CAR constructs, as well as a variety of vectors and cells. The customization process can be completed with just a few simple clicks, please feel free to try it out.
CAR and CAR Cell Customizing System
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For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.
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