Anti-CD72 (4G4) 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
CD72

Targeting Cell Type
T Cell

Targeting Diseases
Breast 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

Customize Your CAR Products

Published Data

Complete CAR data Cyt

Fig.1 In vitro cytotoxicity of CAR cells.

CAR Construction : NbD4-41BB-CD3ζ

Fig.1 In vitro cytotoxicity of CAR cells.

Three B-ALL cell lines (SEM, RS411, and NALM6) and three lymphoma cell lines (TOLEDO-DLBCL, Namalwa-Burkitt lymphoma, and JEKO-mantle cell lymphoma) evaluated susceptibility to either CD19 or CD72-directed CAR T cytotoxicity.

Nix, M. A., Mandal, K., Geng, H., Paranjape, N., Lin, Y. H. T., Rivera, J. M., ... & Wiita, A. P. (2021). Surface Proteomics Reveals CD72 as a Target for In Vitro–Evolved Nanobody-Based CAR-T Cells in KMT2A/MLL1-Rearranged B-ALLCD72 Nanobody-Based CAR-T in MLLr B-ALL. Cancer discovery, 11(8), 2032-2049.

Complete CAR data FuncS

Fig.2 Determination of cytokines release.

CAR Construction : NbD4-41BB-CD3ζ

Fig.2 Determination of cytokines release.

Cytokines released by different CD8 CAR-T cells after 1:1 coculture for 24 hours with the SEM B-ALL cell line.

Complete CAR data FCM

Fig.3 CAR-T cell proliferation assay.

CAR Construction : NbD4-41BB-CD3ζ

Fig.3 CAR-T cell proliferation assay.

Proliferation of CD4 and CD8 CAR-T cells cultured alone or at a 1:1 E:T ratio with either AMO1 tumor cells (multiple myeloma, CD19- and CD72- ) or SEM tumor cells (B-ALL, CD19+ and CD72+).

Complete CAR data BI

Fig.4 CD72 CAR-T cells eradicate tumors and prolong survival in cell line and xenograft models of B-ALL.

CAR Construction : NbD4-41BB-CD3ζ

Fig.4 CD72 CAR-T cells eradicate tumors and prolong survival in cell line and xenograft models of B-ALL.

A: MLLr B-ALL PDX and were treated on day 10 with different CAR-T cells; B, SEM B-ALL cells, treated on day 3 with different CAR-T cells; C, CD19-negative SEM B-ALL cells, treated on day 3 with different CAR-T cells.

CAR scFv data SPR

Fig.5 The binding affinity measurement of NbD4 to CD72.

CAR Construction :

Fig.5 The binding affinity measurement of NbD4 to CD72.

Biolayer interferometry demonstrated that NbD4 bound with surprisingly low affinity to recombinant CD72 (K D~800 nM), with both slow on rate (k on 8.38e4 M-1s-1) and rapid off rate (k off 6.82e-2s -1).

Nix, M. A., Temple, W. C., Karlon, W., Wang, D., Phojanakong, P., Steri, V., ... & Wiita, A. P. (2021). CD72 Nanobody-Based CAR-T Cells Have Potent Anti-Tumor Efficacy in B Cell Malignancies. Blood, 138, 1717.

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.