Anti-GPC3 ( 4G7) h(CD28-CD3ζ) CAR, pCDCAR1 vector

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

Details

Target
GPC3

Targeting Cell Type
T cell

Targeting Diseases
hepatocellular carcinoma

Generation
Second

Vector Name
pCDCAR1

Vector Length
~8kb

Vector Type
Lentiviral vector

Receptor Construction
scFv-CD28-CD3ζ

Discription of 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.

Target

Customize Your CAR Products

Published Data

Complete CAR data FuncS

Fig.1 IL-2 release analysis of 4G7 CAR constructs co-clutured with target cell.

CAR Construction : 1G4 scfv-CD28-CD3ζ

Fig.1 IL-2 release analysis of 4G7 CAR constructs co-clutured with target cell.

CD19 CAR Jurkat (2e5 cells) co-cultured with 2e4 NALM-6 cells for 21 h. IL-2 level secreted by CD19 CAR Jurkat was measured through ELISA assay.

Kang, C. H., Kim, Y., Lee, H. K., Lee, S. M., Jeong, H. G., Choi, S. U., & Park, C. H. (2020). Identification of potent CD19 scFv for CAR T cells through scFv screening with NK/T-cell line. International Journal of Molecular Sciences, 21(23), 9163.

Complete CAR data FuncS

Fig.2 Tumor-killing ability of eight different 4G7-CAR

CAR Construction : 1G4 scfv-CD28-CD3ζ

Fig.2 Tumor-killing ability of eight different 4G7-CAR

Tumor-killing ability of eight different CD19 CAR KHYG-1 cells against luciferase-expressing NALM-6. CD19 CAR KHYG-1 cells (E:T ratio 3:1 or 10:1) were co-cultured with luciferase-expressing NALM-6 (1.5 × 104 cells) for 7 h.

Complete CAR data FuncS

Fig.3 4G7 CAR constructs effectively recognize antigen CD19.

CAR Construction : 1G4 scfv-CD28-CD3ζ

Fig.3 4G7 CAR constructs effectively recognize antigen CD19.

FMC63 CAR KHYG-1 or 4G7 CAR KHYG-1 cells were incubated with NALM-6 at different E:T ratios (1:1, 3:1, and 10:1) and different incubation times (6 h and 23 h) to see their lytic activity.

Complete CAR data FuncS

Fig.4 FMC63 and 4G7 CAR T cells effectively responded to CD19-positive NALM-6 cells.

CAR Construction : 1G4 scfv-CD28-CD3ζ

Fig.4 FMC63 and 4G7 CAR T cells effectively responded to CD19-positive NALM-6 cells.

Tumor-killing ability of FMC63, 4G7, and FITC CAR T cells against luciferase-expressing NALM-6 and U937. FMC63, 4G7, and FITC CAR T cells (2 × 105 cells) were co-cultured with luciferase-expressing NALM-6 (2 × 104 cells) or U937 (2 × 104 cells) for indicated incubation times at the E:T ratio 10:1. Then, luminescence values were measured.

Complete CAR data FuncS

Fig.5 Cytokine release analysis of 4G7 CAR constructs co-clutured with target cell.

CAR Construction : 1G4 scfv-CD28-CD3ζ

Fig.5 Cytokine release analysis of 4G7 CAR constructs co-clutured with target cell.

FMC63, 4G7, or FITC CAR T cells (1.5 × 105 cells) were co-cultured with NALM-6 cells (1.5 × 104 cells) or U937 cells (1.5 × 104 cells) for 21 h at the E:T ratio 10:1. The amount of IFN-γ and IL-2 secreted by CAR T cells was measured through the ELISA assay.

Complete CAR data FuncS

Fig.6 4G7 CAR T cells eliminated acute-lymphoblastic-leukemia-cell NALM-6 in vivo.

CAR Construction : 1G4 scfv-CD28-CD3ζ

Fig.6 4G7 CAR T cells eliminated acute-lymphoblastic-leukemia-cell NALM-6 in vivo.

NSG mice were injected intravenously (i.v.) with 5E6 Luc-expressing NALM-6 cells. Next day, mice were administered with 1E7 4G7 CAR T cells or phosphate buffered saline (PBS) control (i.v.). From day 5, tumor progression was observed via bioluminescence imaging.

Complete CAR data FuncS

Fig.7 4G7 CAR T cells eliminated acute-lymphoblastic-leukemia-cell NALM-6 in vivo.

CAR Construction : 1G4 scfv-CD28-CD3ζ

Fig.7 4G7 CAR T cells eliminated acute-lymphoblastic-leukemia-cell NALM-6 in vivo.

Luminescence value of each group of in vivo.

Complete CAR data FuncS

Fig.8 4G7 CAR T cells eliminated acute-lymphoblastic-leukemia-cell NALM-6 in vivo.

CAR Construction : 1G4 scfv-CD28-CD3ζ

Fig.8 4G7 CAR T cells eliminated acute-lymphoblastic-leukemia-cell NALM-6 in vivo.

Percentage survival was calculated of in vivo.

More Published Data More Published Data

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.