Anti-EGFR (806) h(CD28-CD3ζ) CAR, pCDCAR1 vector

All products and services are For Research Use Only and CANNOT be used in the treatment or diagnosis of disease.

Details

Target
EGFR

Targeting Cell Type
T cell

Targeting Diseases
Melanoma

Generation
Second

Vector Name
pCDCAR1

Vector Length
~8kb

Vector Type
Lentiviral

Receptor Construction
scFv-CD28-CD3ζ

Discription of Signaling Cassetes
CD28
CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide co-stimulatory signals 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.
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

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Published Data

CAR scFv data FCM

Fig.1 FACS analysis of parental stably expressing wt (U87MG.wtEGFR).

CAR Construction :

Fig.1 FACS analysis of parental stably expressing wt (U87MG.wtEGFR).

The negative control (irrelevant antibody) fluorescence is plotted on each panel (black line).

Panousis, C., Rayzman, V. M., Johns, T. G., Renner, C., Liu, Z., Cartwright, G., ... & Scott, A. M. (2005). Engineering and characterisation of chimeric monoclonal antibody 806 (ch806) for targeted immunotherapy of tumours expressing de2-7 EGFR or amplified EGFR. British journal of cancer, 92(6), 1069-1077.

CAR scFv data Cyt

Fig.2 Immune effector function of mAb 806.

CAR Construction :

Fig.2 Immune effector function of mAb 806.

Representative results from three separate experiments are shown.

Complete CAR data FuncS

Fig.3 806 CAR T cells exhibit low or no affinity for EGFR expressed on primary astrocytes and keratinocytes.

CAR Construction : 806 scFv-28ζ

Fig.3 806 CAR T cells exhibit low or no affinity for EGFR expressed on primary astrocytes and keratinocytes.

Individual bioluminescence was assessed in 806 and 2173 CAR T cell treated groups.

Thokala, R., Binder, Z. A., Yin, Y., Zhang, L., Zhang, J. V., Zhang, D. Y., ... & O’Rourke, D. M. (2021). High-Affinity Chimeric Antigen Receptor With Cross-Reactive scFv to Clinically Relevant EGFR Oncogenic Isoforms. Frontiers in Oncology, 11.

Complete CAR data FuncS

Fig.4 Anti-tumor activity of 806 CAR T cells in in vivo.

CAR Construction : 806 scFv-28ζ

Fig.4 Anti-tumor activity of 806 CAR T cells in in vivo.

Individual bioluminescence was assessed in 806 and 2173 CAR T cell treated groups

Complete CAR data IHC

Fig.5 High fidelity GBM organoids demonstrate cross-reactivity of 806 CAR.

CAR Construction : 806 scFv-28ζ

Fig.5 High fidelity GBM organoids demonstrate cross-reactivity of 806 CAR.

wtEGFR staining in both GBO lines provided additional evidence of the cross-reactive nature of the 806 CAR.

Complete CAR data Cyt

Fig.6 In vitro characterization of 806 EGFR CAR T cells.

CAR Construction : 806 scFv-28ζ

Fig.6 In vitro characterization of 806 EGFR CAR T cells.

Antigen specific cytolytic activity of 806 and 2173 CAR T cells against cell lines expressing EGFR and its variants.

More Published Data More Published Data

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.

pCDCAR1 EGFR h(28ζ) (CAR-LC098)

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