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Anti-CD19 (4G7) h(41BB-CD3ζ) Multi-chain CAR, pCDCAR1 (CAR-LC296)


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

The vector of anti-CD19 chimeric antigen receptor (CAR) is based on the FcεRI receptor scaffold and constructed for the engineering of T cells to target human CD19. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-CD19 antibody linked to FcRα, FcRγ, and FcRβ linked to CD137 (4-1BB) and CD3ζ signaling domains. And the vector product was designed for the treatment of chronic lymphoid leukemia.

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Details

  • Target
  • CD19
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Chronic lymphocytic leukemia
  • Generation
  • Third
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Lentiviral
  • Receptor Construction
  • FcRα-scFv-CD8/FcRβ-ΔITAM-41BB-CD3ζ/FcRγ
  • Discription of Signaling Cassetes
  • 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 CD28 and OX40 can be used with CD3-zeta to transmit a proliferative/survival signal, or all three can be used together.

Target

  • Clone
  • 4G7
  • Host
  • Mouse
  • Target Species
  • Human
  • Gene Name
  • CD19
  • Synonyms
  • CD19;CD19; CD19 Molecule; CD19 Molecule; B-Lymphocyte Surface Antigen B4; T-Cell Surface Antigen Leu-12; Differentiation Antigen CD19; CD19 Antigen; B-Lymphocyte Antigen CD19; CVID3; B4;

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  • 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 Latest CAR Construction

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 Latest CAR Construction

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.

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.3 4G7 CAR constructs effectively recognize antigen CD19.

CAR Construction : 1G4 scfv-CD28-CD3ζ Latest CAR Construction

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.

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

CAR Construction : 1G4 scfv-CD28-CD3ζ Latest CAR Construction

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.

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

CAR Construction : 1G4 scfv-CD28-CD3ζ Latest CAR Construction

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.

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

CAR Construction : 1G4 scfv-CD28-CD3ζ Latest CAR Construction

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.

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

CAR Construction : 1G4 scfv-CD28-CD3ζ Latest CAR Construction

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

Luminescence value of each group of in vivo.

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

CAR Construction : 1G4 scfv-CD28-CD3ζ Latest CAR Construction

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

Percentage survival was calculated of in vivo.

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.

More Published Data More Published Data

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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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