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Anti-HER2 (4D5) h(41BB-CD3ζ) CAR, pCDCAR1 (CAR-LC172)


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

The vector of anti-HER2 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human HER2. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-HER2 antibody linked to 4-1BB (CD137) and CD3ζ signaling domains. And the vector product was designed for the treatment of breast carcinomas.

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Details

  • Target
  • HER2
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Breast carcinomas
  • Generation
  • Second
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Lentiviral
  • 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
  • 4D5
  • Host
  • Mouse
  • Target Species
  • Human
  • Gene Name
  • erb-b2 receptor tyrosine kinase 2
  • Synonyms
  • NEU; NGL; HER2; TKR1; CD340; HER-2; MLN 19; HER-2/neu

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  • Published Data
Complete CAR data ELISA

Fig.1 Cytokine release assay.

CAR Construction : 4D5-4-1BB-CD3ζ Latest CAR Construction

Fig.1 Cytokine release assay.

HER2-directed CAR-T cells specifically secreted IFN-γ and TNF-α when cultured with HER2+ tumor cells.

Zhang, P. F., Huang, Y., Liang, X., Li, D., Jiang, L., Yang, X., ... & Wang, W. (2020). Enhancement of the antitumor effect of HER2‐directed CAR‐T cells through blocking epithelial‐mesenchymal transition in tumor cells. The FASEB Journal, 34(8), 11185-11199.

Complete CAR data BI

Fig.2 Antitumor efficacy of HER2-directed CAR-T cells in vivo.

CAR Construction : 4D5-4-1BB-CD3ζ Latest CAR Construction

Fig.2 Antitumor efficacy of HER2-directed CAR-T cells in vivo.

In vivo live imaging of the metastatic ovarian cancer model. The bioluminescent signals were lower in the CAR-T cells combined with A83-01 group than in other groups.

Zhang, P. F., Huang, Y., Liang, X., Li, D., Jiang, L., Yang, X., ... & Wang, W. (2020). Enhancement of the antitumor effect of HER2‐directed CAR‐T cells through blocking epithelial‐mesenchymal transition in tumor cells. The FASEB Journal, 34(8), 11185-11199.

Complete CAR data FuncS

Fig.3 Kaplan-Meier analysis of the overall survival for each group.

CAR Construction : 4D5-4-1BB-CD3ζ Latest CAR Construction

Fig.3 Kaplan-Meier analysis of the overall survival for each group.

CAR-T cells combined with A83-01 significantly prolonged the survival time of mice in the CAR T+ A83-01 group and significantly prolonged survival compared with all other treatment group.

Zhang, P. F., Huang, Y., Liang, X., Li, D., Jiang, L., Yang, X., ... & Wang, W. (2020). Enhancement of the antitumor effect of HER2‐directed CAR‐T cells through blocking epithelial‐mesenchymal transition in tumor cells. The FASEB Journal, 34(8), 11185-11199.

Complete CAR data ELISA

Fig.4 Cytokine production of 4D5 CAR-transduced T cells.

CAR Construction : 4D5-CD28-CD3ζ Latest CAR Construction

Fig.4 Cytokine production of 4D5 CAR-transduced T cells.

IFN-γ secretion of 4D5-28Z-transduced T cells following coculture with the indicated tumor lines.

Zhao, Y., Wang, Q. J., Yang, S., Kochenderfer, J. N., Zheng, Z., Zhong, X., ... & Morgan, R. A. (2009). A herceptin-based chimeric antigen receptor with modified signaling domains leads to enhanced survival of transduced T lymphocytes and antitumor activity. The Journal of Immunology, 183(9), 5563-5574.

Complete CAR data FuncS

Fig.5 Specific tumor cell lysis by 4D5 CAR-transduced T cells.

CAR Construction : 4D5-CD28-CD3ζ Latest CAR Construction

Fig.5 Specific tumor cell lysis by 4D5 CAR-transduced T cells.

4D5 CAR-transduced PBLs (4D5-28Z) were tested in a 51Cr release assay, where nontransduced PBLs (NV) served as a control.

Zhao, Y., Wang, Q. J., Yang, S., Kochenderfer, J. N., Zheng, Z., Zhong, X., ... & Morgan, R. A. (2009). A herceptin-based chimeric antigen receptor with modified signaling domains leads to enhanced survival of transduced T lymphocytes and antitumor activity. The Journal of Immunology, 183(9), 5563-5574.

Complete CAR data FuncS

Fig.6 Treatment of a breast cancer xenograft by ErbB2-CAR-transduced PBLs.

CAR Construction : 4D5-CD28-CD3ζ, 4D5-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.6 Treatment of a breast cancer xenograft by ErbB2-CAR-transduced PBLs.

Adoptively transferred (i.v.) 4D5-28Z or 4D5-CD8-28BBZ- or SP6-28Z-transduced PBLs were administered to animals bearing human breast tumor.

Zhao, Y., Wang, Q. J., Yang, S., Kochenderfer, J. N., Zheng, Z., Zhong, X., ... & Morgan, R. A. (2009). A herceptin-based chimeric antigen receptor with modified signaling domains leads to enhanced survival of transduced T lymphocytes and antitumor activity. The Journal of Immunology, 183(9), 5563-5574.

Complete CAR data ELISA

Fig.7 HER2-CAR T cells administered intrathecally to non-human primates (NHP) did not cause toxicity.

CAR Construction : 4D5-4-1BB-CD3ζ Latest CAR Construction

Fig.7 HER2-CAR T cells administered intrathecally to non-human primates (NHP) did not cause toxicity.

Summary of CSF cytokine levels after infusion of T-HER2 target cells and HER2 CAR T cells.

Nellan, A., Rota, C., Majzner, R., Lester-McCully, C. M., Griesinger, A. M., Mulcahy Levy, J. M., ... & Lee, D. W. (2018). Durable regression of Medulloblastoma after regional and intravenous delivery of anti-HER2 chimeric antigen receptor T cells. Journal for immunotherapy of cancer, 6(1), 1-14.

Complete CAR data FCM

Fig.8 Interferon gamma (IFN-γ) release.

CAR Construction : 4D5-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.8 Interferon gamma (IFN-γ) release.

CAR-T cells were co-cultured with MDA.MB.468, MDA.MB.231, SKOV3, or MDA.MB.468.Her2 cells with Brefeldin A protein transport
inhibitor for 6 h to detect interferon gamma (IFN-c) release.

Siegler, E., Li, S., Kim, Y. J., & Wang, P. (2017). Designed ankyrin repeat proteins as Her2 targeting domains in chimeric antigen receptor-engineered T cells. Human gene therapy, 28(9), 726-736.

Complete CAR data FCM

Fig.9 Cytotoxicity assay.

CAR Construction : 4D5-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.9 Cytotoxicity assay.

CAR-T cells were co-cultured for 24 h with MDA.MB.468, MDA.MB.231,SKOV3, or MDA.MB.468.Her2 target cells at 1:1, 3:1, 5:1, or 10:1 effector-to-target ratios.

Siegler, E., Li, S., Kim, Y. J., & Wang, P. (2017). Designed ankyrin repeat proteins as Her2 targeting domains in chimeric antigen receptor-engineered T cells. Human gene therapy, 28(9), 726-736.

Complete CAR data FuncS

Fig.10 4D5 CAR-T cells inhibit tumor growth in vivo.

CAR Construction : 4D5-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.10 4D5 CAR-T cells inhibit tumor growth in vivo.

Mice bearing SKOV3 cells were treated with 10 million NT, 4D5, G3, or 929 CAR-T cells on day 14 via tail-vein injection.

Siegler, E., Li, S., Kim, Y. J., & Wang, P. (2017). Designed ankyrin repeat proteins as Her2 targeting domains in chimeric antigen receptor-engineered T cells. Human gene therapy, 28(9), 726-736.

CAR scFv data SPR

Fig.11 Binding kinetics of the 4D5 antibodies.

CAR Construction : Latest CAR Construction

Fig.11 Binding kinetics of the 4D5 antibodies.

The increase of functional affinity obtained by multimerization was measured by sur-face plasmon resonance (BIAcore) on recombinant p185HER-2-ECD.

Willuda, J., Kubetzko, S., Waibel, R., Schubiger, P. A., Zangemeister-Wittke, U., & Plückthun, A. (2001). Tumor targeting of mono-, di-, and tetravalent anti-p185HER-2 miniantibodies multimerized by self-associating peptides. Journal of Biological Chemistry, 276(17), 14385-14392.

CAR scFv data SPR

Fig.12 Binding kinetics of the 4D5 antibodies.

CAR Construction : Latest CAR Construction

Fig.12 Binding kinetics of the 4D5 antibodies.

The increase of functional affinity obtained by multimerization was measured on living SK-OV-3 cells overexpressing p185HER-2.

Willuda, J., Kubetzko, S., Waibel, R., Schubiger, P. A., Zangemeister-Wittke, U., & Plückthun, A. (2001). Tumor targeting of mono-, di-, and tetravalent anti-p185HER-2 miniantibodies multimerized by self-associating peptides. Journal of Biological Chemistry, 276(17), 14385-14392.

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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