Anti-CEACAM5 (hMN-14) 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
CEACAM5

Targeting Cell Type
T cell

Targeting Diseases
Colorectal cancer

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 costimulatory 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 Histograms of antibody binding to carcinoembryonic antigen (CEA) expressing MKN-45.

CAR Construction :

Fig.1 Histograms of antibody binding to carcinoembryonic antigen (CEA) expressing MKN-45.

The secondary antibody only served as controls (gray-filled histograms).

Shinmi, D., Nakano, R., Mitamura, K., Suzuki‐Imaizumi, M., Iwano, J., Isoda, Y., ... & Masuda, K. (2017). Novel anticarcinoembryonic antigen antibody–drug conjugate has antitumor activity in the existence of soluble antigen. Cancer Medicine, 6(4), 798-808.

CAR scFv data FuncS

Fig.2 Binding activity of anti-carcinoembryonic antigen (CEA) antibodies to membrane-bound CEA on MKN-45 under the presence or
absence of soluble CEA.

CAR Construction :

Fig.2 Binding activity of anti-carcinoembryonic antigen (CEA) antibodies to membrane-bound CEA on MKN-45 under the presence or absence of soluble CEA.

All measurements were performed in triplicate.

CAR scFv data Cyt

Fig.3 In vitro cytotoxicity of 15-1-32-vcMMAE (open circle), labetuzumab-vcMMAE (closed circle), naked 15-1-32 (open square),
naked labetuzumab (closed square) on carcinoembryonic antigen (CEA)-positive gastric cancer cell lines, MKN-45.

CAR Construction :

Fig.3 In vitro cytotoxicity of 15-1-32-vcMMAE (open circle), labetuzumab-vcMMAE (closed circle), naked 15-1-32 (open square), naked labetuzumab (closed square) on carcinoembryonic antigen (CEA)-positive gastric cancer cell lines, MKN-45.

NT stands for not antibody treated.

Complete CAR data FuncS

Fig.4 IFN-γ quantitation in the medium at 12 and 24 h after coculture of short spacer CEACAM5 CAR-transduced.

CAR Construction : hMN-14 scFv-28ζ

Fig.4 IFN-γ quantitation in the medium at 12 and 24 h after coculture of short spacer CEACAM5 CAR-transduced.

Data are representative of three independent experiments with similar results. ns represents nonsignificance. ****P < 0.0001 (by two-way ANOVA statistical analysis).

Lee, J. K., Bangayan, N. J., Chai, T., Smith, B. A., Pariva, T. E., Yun, S., ... & Witte, O. N. (2018). Systemic surfaceome profiling identifies target antigens for immune-based therapy in subtypes of advanced prostate cancer. Proceedings of the National Academy of Sciences, 115(19), E4473-E4482.

Complete CAR data ELISA

Fig.5 Relative viability over time of CEACAM5-negative MSKCC EF1 target cells or CEACAM5-positive NCI-H660 target cells cocultured with long spacer CEACAM5 CAR-transduced T cells.

CAR Construction : hMN-14 scFv-28ζ

Fig.5 Relative viability over time of CEACAM5-negative MSKCC EF1 target cells or CEACAM5-positive NCI-H660 target cells cocultured with long spacer CEACAM5 CAR-transduced T cells.

Effector-to-target ratios varying from 1:5 to 2:1 are shown. SE measurements for three replicate wells at each timepoint are displayed. Data are representative of two independent experiments with similar results.

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.