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Anti-EGFR (Nimotuzumab) h(41BB-CD3ζ) CAR, pCDCAR1 (CAR-MZ011)

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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-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.

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Details

  • Target
  • EGFR
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Human primary glioblastoma
  • 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
  • Nimotuzumab
  • Host
  • Humanized
  • 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 SPR

Fig.1 SPR/Biacore experiments showing the inhibition of EGF binding to eEGFR with increasing amounts of Nimotuzumab.

CAR Construction : Latest 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 scFv data FCM

Fig.2 Recognition of the EGFR chimera by Nimotuzumab, measured by FACS.

CAR Construction : Latest 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.

Complete CAR data FuncS

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.

CAR Construction : Nimotuzumab scFv-28ζ Latest CAR Construction

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.

Complete CAR data FuncS

Fig.4 Representative histograms of expression of tEGFR on EL4 cells relative to cell lines negative for EGFR.

CAR Construction : Nimotuzumab scFv-28ζ Latest CAR Construction

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.

Complete CAR data FCM

Fig.5 Representative histograms of Intracellular and surface expression of CAR determined by flow cytometry.

CAR Construction : Nimotuzumab scFv-28ζ Latest CAR Construction

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.

Complete CAR data FuncS

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).

CAR Construction : Nimotuzumab scFv-28ζ Latest CAR Construction

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.

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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