Anti-EGFR (Matuzumab) h(4-1BB-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 vector

Receptor Construction
scFv-4-1BB-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 antigennonspecific 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

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Published Data

CAR scFv data FCM

Fig.1 EGFR expression detected by Matuzumab on FACS analysis of A431, Caski and C33A cells.

CAR Construction :

Fig.1 EGFR expression detected by Matuzumab on FACS analysis of A431, Caski and C33A cells.

Student's t test * P < 0.05, when compared to controls.

Meira, D. D., Almeida, V. H., Mororó, J. S., Caetano, M. S., Nóbrega, I. P., Batista, D., ... & Ferreira, C. G. (2011). Efficient Blockade of Akt signaling is a determinant factor to overcome resistance to Matuzumab. Molecular cancer, 10(1), 1-13.

CAR scFv data FCM

Fig.2 Combination of matuzumab and gemcitabine induces tumor cell apoptosis and cell cycle arrest in vitro.

CAR Construction :

Fig.2 Combination of matuzumab and gemcitabine induces tumor cell apoptosis and cell cycle arrest in vitro.

All measurements were performed in triplicate.

Kleespies, A., Ischenko, I., Eichhorn, M. E., Seeliger, H., Amendt, C., Mantell, O., ... & Bruns, C. J. (2008). Matuzumab short-term therapy in experimental pancreatic cancer: prolonged antitumor activity in combination with gemcitabine. Clinical Cancer Research, 14(17), 5426-5436.

CAR scFv data ELISA

Fig.3 Binding of mAbs cetuximab and matuzumab to peptide arrays.

CAR Construction :

Fig.3 Binding of mAbs cetuximab and matuzumab to peptide arrays.

Data are represented as mean±s.d.

Hartmann, C., Müller, N., Blaukat, A., Koch, J., Benhar, I., & Wels, W. S. (2010). Peptide mimotopes recognized by antibodies cetuximab and matuzumab induce a functionally equivalent anti-EGFR immune response. Oncogene, 29(32), 4517-4527.

CAR scFv data FCM

Fig.4 Renca-lacZ/EGFR cells were incubated with mAbs cetuximab or matuzumab either without pretreatment, or after pretreatment of cells with a 20-fold molar excess of anti-peptide antibodies or pre-immune serum as indicated.

CAR Construction :

Control cells were treated only with secondary antibody.

CAR scFv data FuncS

Fig.5 Phage binding to scFv fragments of cetuximab or matuzumab were selected in independent sets of experiments from libraries of M13KE bacteriophage displaying randomized linear 7mer, linear 12mer or cyclic 7mer (C7C).

CAR Construction :

Cyclic 7mer (C7C) peptides using IPTG-induced cultures of E. coli HB101 harboring pIB-Tx-scFv(225) or pIB-Tx-scFv(72000).

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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.