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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 CD4 transmembrane domain and FcεRIγ signaling domains. And the vector product was designed for the treatment of glioma.
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CAR Construction :
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. |
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CAR Construction :
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. |
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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. |
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CAR Construction :
Fig.4 Specificity of RAbDMvII1 by Flow cytometry. Freshly dissociated human GBM tumor stained with RAbDMvIII shows 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. |
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