Anti-VAR2CSA (X12X74) h(ICOSBBζ) CAR-Jurkat (XS-1222-LX478)

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All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

The Anti-VAR2CSA (X12X74) h(ICOSBBζ) CAR-Jurkat-NFAT-Luc cell line is a stable cell line derived from anti-VAR2CSA CAR lentivirus transduction using Jurkat-NFAT-luciferase cell as effector cell. This transduced CAR lentiviral vector was constructed to express scFv of an anti-VAR2CSA antibody linked to the ICOS-4-1BB-CD3ζ signaling domains. The CAR-Jurkat cell line constitutively expressing the firefly luciferase gene under the control of NFAT response elements to facilitate subsequent research. And the recombinant cell product targeted VAR2CSA for CAR-T studies in human solid tumors such as melanomas, neuroblastoma.

Specifications

Species
Human

Host Cell Type
T lymphocyte

Overexpressed Gene
NFAT-Luciferase Reporter

Size
Containing ≥ 2 X 10*6 / vial Frozen cells

Culture Condition
Suspension

Growth Pattern
Exponential growth phase

Formulation
Frozen cells

Cell Purity
>90%

Cell Viability
>80%

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).

Warnings
Avoid multiple freeze/thaw cycles

Research Use Only
Our recombinant Jurkat cells 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

Oncogenicity
Positive (In vitro 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-Jurkat 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-Jurkat 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-Jurkat cells or use immediately.
Note: Jurkat 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. Checkpoint inhibitors screening;
5. Safety switches and regulators of CAR-Jurkat functions study;
6. Pre-clinical study in vivo models;
7. CAR-Jurkat signaling, tumor microenvironment;
8. Proof of concept studies for clinical trials;

CAR Design

Target
VAR2CSA

Target Species
Human

scFv Clone
X12X74

scFv Host Spcies
Mouse

CAR Construction
scFv-ICOS-4-1BB-CD3ζ

CAR Signaling Cassetes
ICOS
CD278 or ICOS (Inducible T-cell COStimulator) is a CD28-superfamily costimulatory molecule that is expressed on activated T cells. It is thought to be important for Th2 cells in particular. The protein encoded by this gene belongs to the CD28 and CTLA-4 cell-surface receptor family. It forms homodimers and plays an important role in cell-cell signaling, immune responses, and regulation of cell proliferation.
41BB
CD137 (also known as 4-1BB) is a surface co-stimulatory glycoprotein originally described as present on activated T lymphocytes, which belongs to the tumor necrosis factor (TNF) receptor superfamily. It is expressed mainly on activated CD4+ and CD8+ T cells, and binds to a high-affinity ligand (4-1BBL) expressed on several antigen-presenting cells such as macrophages and activated B cells. On the basis of preclinical observation, this molecule can promote the persistence of antigen-specific and antigen-nonspecific chimeric antigen receptor T-cells to significantly increases antitumor activity.
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 ICOS and 4-1BB can be used with CD3-zeta to transmit a proliferative/survival signal, or all three can be used together.

CAR Vector Name
pCDCAR1

CAR Vector length
~8kb

CAR Vector Type
Lentiviral vector

CAR Generation
Third

Targeting Diseases
solid tumors such as melanomas, neuroblastoma

GeneID
36312543

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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.

Anti-VAR2CSA (X12X74) h(ICOSBBζ) CAR-Jurkat (XS-1222-LX478)

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