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pCDCAR1 EGFRvIII h(ζ) (CAR-YF429)

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

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Details

  • Target
  • EGFRvIII
  • Targeting Cell Type
  • T Cell
  • Targeting Diseases
  • Glioma
  • Generation
  • First
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Lentiviral vector
  • Receptor Construction
  • scFv-CD3ζ
  • Discription of Signaling Cassetes
  • 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 CD21-zeta to transmit a proliferative/survival signal, or all three can be used together.

Target

  • Clone
  • MAb 139
  • Host
  • Human
  • Target Species
  • Human
  • Gene Name
  • Epidermal Growth Factor Receptor
  • Synonyms
  • EGFR;EGFR; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor; Receptor Tyrosine-Protein Kinase ErbB-1; Erb-B2 Receptor Tyrosine Kinase 1; Proto-Oncogene C-ErbB-1; EC 2.7.10.1; ERBB1; ERBB; HER1; Epidermal Growth Factor Receptor (Avian Erythroblastic Leukemia Viral (V-Erb-B) Oncogene Homolog); Erythroblastic Leukemia Viral (V-Erb-B) Oncogene Homolog (Avian);

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  • Published Data
Complete CAR data FCM

Fig.1 EGFRvIII mCAR retroviral gene delivery to murine T cells confers antigen-specific activity.

CAR Construction : MAb 139 scFv-28ζ Latest CAR Construction

Fig.1 EGFRvIII mCAR retroviral gene delivery to murine T cells confers antigen-specific activity.

VM/Dk-derived glioma SMA560vIII shows EGFRvIII expression compared with SMA560 by surface antibody staining (L8A4) and flow cytometry.

Sampson, J. H., Choi, B. D., Sanchez-Perez, L., Suryadevara, C. M., Snyder, D. J., Flores, C. T., ... & Johnson, L. A. (2014). EGFRvIII mCAR-Modified T-Cell Therapy Cures Mice with Established Intracerebral Glioma and Generates Host Immunity against Tumor-Antigen LossEGFRvIII mCARs for Malignant Glioma. Clinical Cancer Research, 20(4), 972-984.

Complete CAR data FuncS

Fig.2 EGFRvIII mCAR T-cell therapy against intracerebral tumors requires lymphodepletive host-conditioning for efficacy, and can be abrogated by soluble peptide administration.

CAR Construction : MAb 139 scFv-28ζ Latest CAR Construction

Fig.2 EGFRvIII mCAR T-cell therapy against intracerebral tumors requires lymphodepletive host-conditioning for efficacy, and can be abrogated by soluble peptide administration.

To evaluate the impact of lymphopenia on the antitumor efficacy of EGFRvIII mCAR T cells, VM/Dk mice (8 per group) with 3-day established intracerebral SMA560vIII tumors.

Sampson, J. H., Choi, B. D., Sanchez-Perez, L., Suryadevara, C. M., Snyder, D. J., Flores, C. T., ... & Johnson, L. A. (2014). EGFRvIII mCAR-Modified T-Cell Therapy Cures Mice with Established Intracerebral Glioma and Generates Host Immunity against Tumor-Antigen LossEGFRvIII mCARs for Malignant Glioma. Clinical Cancer Research, 20(4), 972-984.

Complete CAR data FCM

Fig.3 Treatment of mice bearing EGFRvIIIPOS tumors with mCARs provides long-term protection against EGFRvIIINEG tumor

CAR Construction : MAb 139 scFv-28ζ Latest CAR Construction

Fig.3 Treatment of mice bearing EGFRvIIIPOS tumors with mCARs provides long-term protection against EGFRvIIINEG tumor

SMA560 (EGFRvIIINEG) gliomas were maintained in vitro or passaged in a mouse subcutaneously for 14 days, then harvested and both were stained for EGFRvIII expression using L8A4 mAb and analyzed by flow cytometry.

Sampson, J. H., Choi, B. D., Sanchez-Perez, L., Suryadevara, C. M., Snyder, D. J., Flores, C. T., ... & Johnson, L. A. (2014). EGFRvIII mCAR-Modified T-Cell Therapy Cures Mice with Established Intracerebral Glioma and Generates Host Immunity against Tumor-Antigen LossEGFRvIII mCARs for Malignant Glioma. Clinical Cancer Research, 20(4), 972-984.

Complete CAR data FCM

Fig.4 Development of CARs targeting EGFRvIII. NIH-3T3 cells and BHK cells were transduced with a retroviral vector expressing EGFRvIII and a neomycin resistance gene.

CAR Construction : MAb 139 scFv-28ζ Latest CAR Construction

Fig.4 Development of CARs targeting EGFRvIII. NIH-3T3 cells and BHK cells were transduced with a retroviral vector expressing EGFRvIII and a neomycin resistance gene.

Based on the amino acid sequence for anti-EGFRvIII mAbs, scFv genes were synthesized and fused to T-cell signaling domains from CD28 and CD3f followed by insertion into c-retroviral vector MSGV1 (diagram).

Morgan, R. A., Johnson, L. A., Davis, J. L., Zheng, Z., Woolard, K. D., Reap, E. A., ... & Rosenberg, S. A. (2012). Recognition of glioma stem cells by genetically modified T cells targeting EGFRvIII and development of adoptive cell therapy for glioma. Human gene therapy, 23(10), 1043-1053.

Complete CAR data ELISA

Fig.5 Equivalent function of second- and third-generation anti-EGFRvIII CAR vectors.

CAR Construction : MAb 139 scFv-28ζ Latest CAR Construction

Fig.5 Equivalent function of second- and third-generation anti-EGFRvIII CAR vectors.

Using anti-EGFRvIII human mAb 139, a third-generation CAR vector was assembled using CD28, 4-1BB, and CD3f T-cell signaling domains.

Morgan, R. A., Johnson, L. A., Davis, J. L., Zheng, Z., Woolard, K. D., Reap, E. A., ... & Rosenberg, S. A. (2012). Recognition of glioma stem cells by genetically modified T cells targeting EGFRvIII and development of adoptive cell therapy for glioma. Human gene therapy, 23(10), 1043-1053.

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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