Anti-CEA (Fl1-39) h(CD28-CD3ζ) CAR, pSBCAR1 vector

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

Details

Target
CEA

Targeting Cell Type
T cell

Targeting Diseases
Colon Cancer

Generation
Second

Vector Name
pSBCAR1

Vector Length
~6kb

Vector Type
Sleeping Beauty (SB) transposon

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 ELISA

Fig.2 The CEA binding activity of serially diluted 45scFvLCHα was evaluated by a sandwich ELISA.

CAR Construction :

Fig.2 The CEA binding activity of serially diluted 45scFvLCHα was evaluated by a sandwich ELISA.

Antigen binding activity of isolated 45scFvLCHα.

ARAKAWA, F., HARUNO, M., KUROKI, M., KANDA, H., WATANABE, T., MISUMI, Y., & MATSUOKA, Y. (1993). Construction and Expression of Two Mouse—Human Chimeric Antibodies with High Specificity and Affinity for Carcinoembryonic Antigen. Hybridoma, 12(4), 365-379.

CAR scFv data Inhibit

Fig.3 Competitive inhibition assays of chimeric antibodies against parental MAbs for CEA binding in SPEIAs using purified biotinylated CEA.

CAR Construction :

Fig.3 Competitive inhibition assays of chimeric antibodies against parental MAbs for CEA binding in SPEIAs using purified biotinylated CEA.

Fl 1-39 was dried onto wells of 96-well plates.

CAR scFv data ELISA

Fig.4 Reactivity of mouse MAbs (Fll-35 and Fll-39) with CEA and CEA-related antigens.

CAR Construction :

Fig.4 Reactivity of mouse MAbs (Fll-35 and Fll-39) with CEA and CEA-related antigens.

A SPEIAs using antigen-coated plates and a biotinylated goat anti-human y-chain antibody or a biotinylated horse anti-mouse IgG(H+L) antibody.

CAR scFv data FCM

Fig.1 Detection of the expressed F39scFv/CIR receptors in the transfectoma by flow cytometry.

CAR Construction :

Fig.1 Detection of the expressed F39scFv/CIR receptors in the transfectoma by flow cytometry.

Cell surface expression of the F39scFv CIR receptor protein in the transfectoma was demonstrated by FACS,

Arakawa, F., Shibaguchi, H., Xu, Z. H. O. N. G. W. E. I., & Kuroki, M. (2002). Targeting of T cells to CEA-expressing tumor cells by chimeric immune receptors with a highly specific single-chain anti-CEA activity. Anticancer research, 22(6C), 4285-4289.

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.

Anti-CEA (Fl1-39) h(CD28-CD3ζ) CAR, pSBCAR1 (CAR-SB-LX0161)

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