Anti-PSMA (3D8) 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
PSMA

Targeting Cell Type
T cell

Targeting Diseases
Prostate 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 co-stimulatory 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 cell 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

Complete CAR data FCM

Fig.1 Detection of PSMA cell surface expression on PC3-PSMA cells by 3D8 mAb.

CAR Construction : 3D8 scFv-28ζ

Fig.1 Detection of PSMA cell surface expression on PC3-PSMA cells by 3D8 mAb.

PC3 and PC3-PSMA cellswere analyzedby flowcytometry staining with 3D8 mAb followed by incubation with secondary PE-conjugated anti-mouse IgG antibody (solid line).

Ma, Q., Gomes, E. M., Lo, A. S. Y., & Junghans, R. P. (2014). Advanced generation anti‐prostate specific membrane antigen designer T Cells for prostate cancer immunotherapy. The prostate, 74(3), 286-296.

Complete CAR data FCM

Fig.2 Expression of anti-PSMA CARs in T cells.

CAR Construction : 3D8 scFv-28ζ

Fig.2 Expression of anti-PSMA CARs in T cells.

Expression of IgTCR and IgCD28 TCR.

Complete CAR data WB

Fig.3 Homodimer formation of IgTCR and IgCD28 TCR.

CAR Construction : 3D8 scFv-28ζ

Fig.3 Homodimer formation of IgTCR and IgCD28 TCR.

Positions of monomer and dimer of TCRz, IgTCR and IgCD28 TCR are indicated on the right. Molecular mass markers (kDa) are indicated on the left.

Complete CAR data ELISA

Fig.4 Superior cytokine production by 2ndgen dTc.Non-transduced,IgTCR andIgCD28 TCR transduced T cells were incubated with control PC3 and PC3-PSMA tumor cells for 24 hr.

CAR Construction : 3D8 scFv-28ζ

Fig.4 Superior cytokine production by 2ndgen dTc.Non-transduced,IgTCR andIgCD28 TCR transduced T cells were incubated with control PC3 and PC3-PSMA tumor cells for 24 hr.

Supernatants were harvested and assayed for IL2 and IFNg by ELISA.

Complete CAR data FuncS

Fig.5 Proliferation of dTc in response to PSMAþ tumor cells.

CAR Construction : 3D8 scFv-28ζ

Fig.5 Proliferation of dTc in response to PSMAþ tumor cells.

Total cell proliferation.Non-transduced control,IgTCR and IgCD28TCR transduced T cells with 45% transduction efficiency for both were stimulated with g-irradiated PC3 and PC3-PSMA tumor cells for 3 days,cultured for a further 6 days, and counted on days noted.

Complete CAR data FuncS

Fig.6 GD2 sugar moiety and correlation of anti-GD2 mAb affinity with clinical efficacy.

CAR Construction : 3D8 scFv-28ζ

Fig.6 GD2 sugar moiety and correlation of anti-GD2 mAb affinity with clinical efficacy.

Data are represented as mean±s.d.

Hassani, M., Hajari Taheri, F., Sharifzadeh, Z., Arashkia, A., Hadjati, J., van Weerden, W. M., ... & Abolhassani, M. (2019). Construction of a chimeric antigen receptor bearing a nanobody against prostate a specific membrane antigen in prostate cancer. Journal of Cellular Biochemistry, 120(6), 10787-10795.

Complete CAR data FCM

Fig.7 Schematic presentation of NBPII‐CAR and expression of NBPII‐CAR on electroporated T cells.

CAR Construction : 3D8 scFv-28ζ

Fig.7 Schematic presentation of NBPII‐CAR and expression of NBPII‐CAR on electroporated T cells.

Tumor cells were stained with PE-conjugated anti-PSMA antibody or isotype control antibody and analyzed by flow cytometry.

Complete CAR data ELISA

Fig.8 Recognition of PSMA as target cells.

CAR Construction : 3D8 scFv-28ζ

Fig.8 Recognition of PSMA as target cells.

After 24 hours of coculturing T cells with target cells (PSMA+ LNCaP and PSMA− DU-145 cells at E:T ratios 1:1 and 3:1.

Complete CAR data FuncS

Fig.9 Proliferation of engineered T cells upon encountering PSMA.

CAR Construction : 3D8 scFv-28ζ

Fig.9 Proliferation of engineered T cells upon encountering PSMA.

Data are normalized with the growth rate of negative control cell and reported as a percentage.

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.

pCDCAR1 PSMA h(28ζ) (CAR-ZP027)

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