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pCDCAR1 EPCAM h(BBζ) (CAR-WJ0756)


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

The vector of anti-EPCAM chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target Mouse EPCAM. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-EPCAM antibody linked to 41BB and CD3ζ signaling domains. And the vector product was designed for the treatment of Colorectal cancer.

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Details

  • Target
  • EPCAM
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Colorectal cancer
  • Generation
  • Second
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Lentiviral vector
  • Receptor Construction
  • scFv-41BB-CD3ζ
  • 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
  • G8.8
  • Host
  • Rat
  • Target Species
  • Mouse
  • Gene Name
  • epithelial cell adhesion molecule
  • Synonyms
  • ESA; KSA; M4S1; MK-1; DIAR5; EGP-2; EGP40; KS1/4; MIC18; TROP1; EGP314; HNPCC8; TACSTD1

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  • Published Data
Complete CAR data FCM

Fig.1 Flow cytometry  of EpCAM expression  on the  indicated target cell lines.

CAR Construction : G8.8 scFv-41BB-CD28ζ Latest CAR Construction

Fig.1 Flow cytometry of EpCAM expression on the indicated target cell lines.

3T3 cells do not express EpCAM antigen, while both 4T1 (BALB/c breast cancer cell line) and MC38 (C57BL/6J colon cancer cell line) cells are EpCAM positive.

Qin, D., Li, D., Zhang, B., Chen, Y., Liao, X., Li, X., Alexander, P. B., Wang, Y., & Li, Q. J. (2020). Potential lung attack and lethality generated by EpCAM-specific CAR-T cells in immunocompetent mouse models. Oncoimmunology, 9(1), 1806009.

Complete CAR data Cyt

Fig.2 BALB/c derived EpCAM CAR-T cells specifically kill 4T1 cells in vitro.

CAR Construction : G8.8 scFv-41BB-CD28ζ Latest CAR Construction

Fig.2 BALB/c derived EpCAM CAR-T cells specifically kill 4T1 cells in vitro.

4T1 and 3T3 cell lines were incubated with CAR-T or untransduced control T cells at the indicated E:T ratios for 12 h. CAR-T cytotoxicity was calculated by measuring LDH in the culture medium, which is released by lysed cells. Means of triplicate wells per group are shown. Data were analyzed from two independent experiments and presented as the mean ± SEM.

Qin, D., Li, D., Zhang, B., Chen, Y., Liao, X., Li, X., Alexander, P. B., Wang, Y., & Li, Q. J. (2020). Potential lung attack and lethality generated by EpCAM-specific CAR-T cells in immunocompetent mouse models. Oncoimmunology, 9(1), 1806009.

Complete CAR data FuncS

Fig.3 In vivo activity of mouse EpCAM CAR-T cells.

CAR Construction : G8.8 scFv-41BB-CD28ζ Latest CAR Construction

Fig.3 In vivo activity of mouse EpCAM CAR-T cells.

4T1 tumor volumes after treatment with EpCAM CAR-T or control T cells. BALB/c mice were inoculated with 1 × 105 4T1-luciferase cells on the right flank, after which 3 × 106 EpCAM CAR-T (orange) or control T cells (gray) were intravenously injected into tumor-bearing mice on day 7. Tumor volumes were monitored twice a week (n=5 mice per arm).

Qin, D., Li, D., Zhang, B., Chen, Y., Liao, X., Li, X., Alexander, P. B., Wang, Y., & Li, Q. J. (2020). Potential lung attack and lethality generated by EpCAM-specific CAR-T cells in immunocompetent mouse models. Oncoimmunology, 9(1), 1806009.

Complete CAR data IHC

Fig.4 Alveolar cells express EpCAM and are targeted by EpCAM CAR-T cells.

CAR Construction : G8.8 scFv-41BB-CD28ζ Latest CAR Construction

Fig.4 Alveolar cells express EpCAM and are targeted by EpCAM CAR-T cells.

Representative IHC staining of EpCAM antigen expression of intestine (left) and lung (right) collected on day 3 after CAR-T cell transfer. Scale bar=200 µm (upper row), 100 µm (lower row).

Qin, D., Li, D., Zhang, B., Chen, Y., Liao, X., Li, X., Alexander, P. B., Wang, Y., & Li, Q. J. (2020). Potential lung attack and lethality generated by EpCAM-specific CAR-T cells in immunocompetent mouse models. Oncoimmunology, 9(1), 1806009.

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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