Anti-EGFR (7A6F12) 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
Plasma membrane

Generation
Second

Vector Name
pCDCAR1

Vector Length
~8kb

Vector Type
Lentiviral vector

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 (B7.2) 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 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

Customize Your CAR Products

Published Data

CAR scFv data SPR

Fig.1 SPR analysis of the binding of FcγRIIIa with the glycoengineered cetuximab.

CAR Construction :

Fig.1 SPR analysis of the binding of FcγRIIIa with the glycoengineered cetuximab.

The color curves represent thefitting lines for the analysis of the SPR data.

Giddens, J. P., Lomino, J. V., DiLillo, D. J., Ravetch, J. V., & Wang, L. X. (2018). Site-selective chemoenzymatic glycoengineering of Fab and Fc glycans of a therapeutic antibody. Proceedings of the National Academy of Sciences, 115(47), 12023-12027.

CAR scFv data ELISA

Fig.2 The binding of glycoengineered cetuximab and commercial cetuximab with EGFR on A431 cell.

CAR Construction :

Fig.2 The binding of glycoengineered cetuximab and commercial cetuximab with EGFR on A431 cell.

Data are representative of triplicates.

CAR scFv data FCM

Fig.3 ADCC activity was assessed by measuring the percent CD107a +NK cell populations by FACS.

CAR Construction :

Fig.3 ADCC activity was assessed by measuring the percent CD107a +NK cell populations by FACS.

Data are representative of triplicates.

CAR scFv data Inhibit

Fig.4 Inhibition of EGF-stimulated EGFR phosphorylation by AFM24.

CAR Construction :

Fig.4 Inhibition of EGF-stimulated EGFR phosphorylation by AFM24.

Either AFM24 or comparators (RSV/CD16A scFv-IgAb, EGFR/RSV scFv-IgAb, or cetuximab) and subsequently stimulated with 100 ng/mL recombinant human EGF for 10 min at 37°C.

Wingert, S., Reusch, U., Knackmuss, S., Kluge, M., Damrat, M., Pahl, J., ... & Rajkovic, E. (2021, January). Preclinical evaluation of AFM24, a novel CD16A-specific innate immune cell engager targeting EGFR-positive tumors. In MAbs (Vol. 13, No. 1, p. 1950264). Taylor & Francis.

CAR scFv data ELISA

Fig.5 AFM24 retains high ADCC activity in the presence of competing IgG and at low effector-to-target (E:T) ratio.

CAR Construction :

Fig.5 AFM24 retains high ADCC activity in the presence of competing IgG and at low effector-to-target (E:T) ratio.

One representative experiment out of two is shown for each figure. Error bars represent SD.

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.