All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.
The vector of anti-EGFR chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human EGFR. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-EGFR antibody linked to 41BB and CD3ζ signaling domains. And the vector product was designed for the treatment of Human primary glioblastoma.
CAR Construction : Fig.1 SPR/Biacore experiments showing the inhibition of EGF binding to eEGFR with increasing amounts of Nimotuzumab. Each antibody was added in molar excess to a constant concentration of 600 nmol/L of eEGFR and flowed over the EGF molecules immobilized to a level of 500 response units (RU) on a CM5 chip. Talavera, A., Friemann, R., Gómez-Puerta, S., Martinez-Fleites, C., Garrido, G., Rabasa, A., ... & Moreno, E. (2009). Nimotuzumab, an antitumor antibody that targets the epidermal growth factor receptor, blocks ligand binding while permitting the active receptor conformation. Cancer research, 69(14), 5851-5859. |
CAR Construction : Fig.2 Recognition of the EGFR chimera by Nimotuzumab, measured by FACS. In the dot plots shown in the first three panels, the Y axis corresponds to size scattering. Talavera, A., Friemann, R., Gómez-Puerta, S., Martinez-Fleites, C., Garrido, G., Rabasa, A., ... & Moreno, E. (2009). Nimotuzumab, an antitumor antibody that targets the epidermal growth factor receptor, blocks ligand binding while permitting the active receptor conformation. Cancer research, 69(14), 5851-5859. |
CAR Construction : Nimotuzumab scFv-28ζ Fig.3 Numeric expansion of Cetux-CAR+ and Nimo-CAR+ T cells was determined by calculating fold expansion of CD3+CAR+ T cells, determined by flow cytometry, during each stimulation cycle. Data represented as mean ± SD, n=5. Statistical analysis by two-tailed student's t-test. Caruso, H. G., Hurton, L. V., Najjar, A., Rushworth, D., Ang, S., Olivares, S., ... & Cooper, L. J. (2015). Tuning sensitivity of CAR to EGFR density limits recognition of normal tissue while maintaining potent antitumor activity. Cancer research, 75(17), 3505-3518. |
CAR Construction : Nimotuzumab scFv-28ζ Fig.4 Representative histograms of expression of tEGFR on EL4 cells relative to cell lines negative for EGFR. Nimo-CAR+ T cells have impaired response to low density of EGFR. Caruso, H. G., Hurton, L. V., Najjar, A., Rushworth, D., Ang, S., Olivares, S., ... & Cooper, L. J. (2015). Tuning sensitivity of CAR to EGFR density limits recognition of normal tissue while maintaining potent antitumor activity. Cancer research, 75(17), 3505-3518. |
CAR Construction : Nimotuzumab scFv-28ζ Fig.5 Representative histograms of Intracellular and surface expression of CAR determined by flow cytometry. Data representative of three independent donors. Caruso, H. G., Hurton, L. V., Najjar, A., Rushworth, D., Ang, S., Olivares, S., ... & Cooper, L. J. (2015). Tuning sensitivity of CAR to EGFR density limits recognition of normal tissue while maintaining potent antitumor activity. Cancer research, 75(17), 3505-3518. |
CAR Construction : Nimotuzumab scFv-28ζ Fig.6 Relative tumor growth as assessed by serial BLI of tumor. Significant difference in BLI between mice with no treatment vs. treatment (n=6) with Cetux-CAR+ T cells (n=6, p < 0.01) reached at day 25, two-way ANOVA (Sidak’s post-test). Nimo-CAR+ T cells exhibit impaired targeting of low-density EGFR cells in vivo. Caruso, H. G., Hurton, L. V., Najjar, A., Rushworth, D., Ang, S., Olivares, S., ... & Cooper, L. J. (2015). Tuning sensitivity of CAR to EGFR density limits recognition of normal tissue while maintaining potent antitumor activity. Cancer research, 75(17), 3505-3518. |
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