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Anti-EGFRvIII scFv h(CD28-FcεRIγ) CART, pCDCAR1 (CAR-T-2-M315-2G)


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 CD28 transmembrane domain/ endodomain and FcεRIγ 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
  • Second
  • Vector Name
  • pCDCAR1
  • Vector Length
  • 8kb
  • Vector Type
  • Lentiviral
  • Receptor Construction
  • scFv-CD28-FcεRIγ
  • Discription of Signaling Cassetes
  • CD28
    CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide co-stimulatorysignals 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.
    FcεRIγ
    The high-affinity IgE receptor, also known as FcεRI, or Fc epsilon RI, is the high-affinity receptor for the Fc region of immunoglobulin E (IgE). FcεRI is a key molecule involved in allergic reactions. It is a tetramer composed of 1alpha, 1 beta, and 2 gamma chains. The gamma chains are also subunits of other Fc receptors. The scFv-based CARs engineered to contain a signaling domain from FcεRIγ have been shown to deliver a potent signal for T cell activation and function.

Target

  • Clone
  • RAbDMvIII
  • 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
CAR scFv data ELISA

Fig.1 Calibration curve of the binding of RAbDMvIII and RAbvIII recombinant antibodies to coated EGFRvIII in the ELISA.

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Fig.1 Calibration curve of the binding of RAbDMvIII and RAbvIII recombinant antibodies to coated EGFRvIII in the ELISA.

The concentration range for RAbDMvIII and RAbvIII was 20 to 0.02 ug/ml (x axis) and the absorbance was read at 450 nm (Y axis). The assay was analyzed twice.

Gupta, P., Han, S. Y., Holgado-Madruga, M., Mitra, S. S., Li, G., Nitta, R. T., & Wong, A. J. (2010). Development of an EGFRvIII specific recombinant antibody. BMC biotechnology, 10(1), 1-13.

CAR scFv data FCM

Fig.2 NIH3T3 and HC2 cells were mixed in a 1:1 ratio prior to staining with RAbDMvIII revealing distinct identification of an EGFRvIII cell population.

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Fig.2 NIH3T3 and HC2 cells were mixed in a 1:1 ratio prior to staining with RAbDMvIII revealing distinct identification of an EGFRvIII cell population.

The dot blot (left panel) represents human-IgG1 isotype control.

Gupta, P., Han, S. Y., Holgado-Madruga, M., Mitra, S. S., Li, G., Nitta, R. T., & Wong, A. J. (2010). Development of an EGFRvIII specific recombinant antibody. BMC biotechnology, 10(1), 1-13.

CAR scFv data ELISA

Fig.3 Affinity determination of RAbDMvIII by Saturation ELISA.

CAR Construction : Latest CAR Construction

Fig.3 Affinity determination of RAbDMvIII by Saturation ELISA.

Saturation ELISA shows the Scatchard plot of the binding of RAbDMvIII to EGFRvIII.

Gupta, P., Han, S. Y., Holgado-Madruga, M., Mitra, S. S., Li, G., Nitta, R. T., & Wong, A. J. (2010). Development of an EGFRvIII specific recombinant antibody. BMC biotechnology, 10(1), 1-13.

CAR scFv data FCM

Fig.4 Specificity of RAbDMvII1 by Flow cytometry.

CAR Construction : Latest CAR Construction

Fig.4 Specificity of RAbDMvII1 by Flow cytometry.

Freshly dissociated human GBM tumor stained with RAbDMvIII shows
the presence of surface EGFRvIII on 13% of cells (right panel).

Gupta, P., Han, S. Y., Holgado-Madruga, M., Mitra, S. S., Li, G., Nitta, R. T., & Wong, A. J. (2010). Development of an EGFRvIII specific recombinant antibody. BMC biotechnology, 10(1), 1-13.

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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