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Anti-PSCA (Ha1-4.117 ) h(CD3ζ) CAR, pCDCAR1 (CAR-MZ038)


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

The vector of anti-PSCA chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human PSCA. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-PSCA antibody linked to CD3ζ signaling domains. And the vector product was designed for the treatment of Pancreatic cancer.

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Details

  • Target
  • PSCA
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Pancreatic cancer
  • Generation
  • First
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Lentiviral
  • Receptor Construction
  • scFv-CD3ζ
  • Discription of Signaling Cassetes
  • 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

  • Clone
  • Ha1-4.117
  • Host
  • Human
  • Target Species
  • Human
  • Gene Name
  • prostate stem cell antigen
  • Synonyms
  • PSCA;PSCA; prostate stem cell antigen; PRO232;

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  • Published Data
Complete CAR data FCM

Fig.1 FACS analysis of CAR expression in primary human T cells. OKT3-stimulated peripheral blood mononuclear cells were transduced twice with retroviral vectors encoding the indicated CAR constructs.

CAR Construction : Ha1-4.117 scFv-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.1 FACS analysis of CAR expression in primary human T cells. OKT3-stimulated peripheral blood mononuclear cells were transduced twice with retroviral vectors encoding the indicated CAR constructs.

A Herceptin-based anti-Her2/neu CAR was used as a positive control. SA-PE: cells incubated with secondary staining (streptavidin-PE) alone as isotype control.

Abate-Daga, D., Lagisetty, K. H., Tran, E., Zheng, Z., Gattinoni, L., Yu, Z., ... & Morgan, R. A. (2014). A novel chimeric antigen receptor against prostate stem cell antigen mediates tumor destruction in a humanized mouse model of pancreatic cancer. Human gene therapy, 25(12), 1003-1012.

Complete CAR data FCM

Fig.2 Surface expression of anti-PSCA and anti-MSLN CARs.

CAR Construction : Ha1-4.117 scFv-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.2 Surface expression of anti-PSCA and anti-MSLN CARs.

Histograms showing results of protein-L staining, analyzed by FACS, of primary human T lymphocytes engineered to express the two
versions of Ha1-4.117-based anti-PSCA CARs or the antiMSLN CAR.

Abate-Daga, D., Lagisetty, K. H., Tran, E., Zheng, Z., Gattinoni, L., Yu, Z., ... & Morgan, R. A. (2014). A novel chimeric antigen receptor against prostate stem cell antigen mediates tumor destruction in a humanized mouse model of pancreatic cancer. Human gene therapy, 25(12), 1003-1012.

Complete CAR data IHC

Fig.3 Immunohistochemistry staining of PSCA CAR-treated tumors.

CAR Construction : Ha1-4.117 scFv-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.3 Immunohistochemistry staining of PSCA CAR-treated tumors.

Immunohistochemistry of tumor sections showing strong PSCA staining in control groups and greatly reduced staining in PSCA CAR-treated mice. Scale bars represent 2000 lm in the top row and 200 lm in the bottom row.

Abate-Daga, D., Lagisetty, K. H., Tran, E., Zheng, Z., Gattinoni, L., Yu, Z., ... & Morgan, R. A. (2014). A novel chimeric antigen receptor against prostate stem cell antigen mediates tumor destruction in a humanized mouse model of pancreatic cancer. Human gene therapy, 25(12), 1003-1012.

Complete CAR data FuncS

Fig.4 PSCA CARs effectively treat established tumors in NSG mice.

CAR Construction : Ha1-4.117 scFv-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.4 PSCA CARs effectively treat established tumors in NSG mice.

Growth curves after treatment with CARexpressing human T cells or controls, in two independent experiments.

Abate-Daga, D., Lagisetty, K. H., Tran, E., Zheng, Z., Gattinoni, L., Yu, Z., ... & Morgan, R. A. (2014). A novel chimeric antigen receptor against prostate stem cell antigen mediates tumor destruction in a humanized mouse model of pancreatic cancer. Human gene therapy, 25(12), 1003-1012.

Complete CAR data FCM

Fig.5 Flow cytometry analysis of CD69 and CD25 abundance on preinfusion (day 12) CD8+ T cells that expressed GFP or CARs, as indicated.

CAR Construction : Ha1-4.117 scFv-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.5 Flow cytometry analysis of CD69 and CD25 abundance on preinfusion (day 12) CD8+ T cells that expressed GFP or CARs, as indicated.

Histograms are representative of 4 independent experiments.

Ramello, M. C., Benzaïd, I., Kuenzi, B. M., Lienlaf-Moreno, M., Kandell, W. M., Santiago, D. N., ... & Abate-Daga, D. (2019). An immunoproteomic approach to characterize the CAR interactome and signalosome. Science signaling, 12(568), eaap9777.

Complete CAR data WB

Fig.6 Western blot analysis of CD3ζ interaction in lysates of T cells
transduced with PSCA-28t28Z, PSCA-8t28BBZ, or GFP and immunoprecipitated for the
CAR scFv.

CAR Construction : Ha1-4.117 scFv-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.6 Western blot analysis of CD3ζ interaction in lysates of T cells transduced with PSCA-28t28Z, PSCA-8t28BBZ, or GFP and immunoprecipitated for the CAR scFv.

Data are represented as mean±s.d.

Ramello, M. C., Benzaïd, I., Kuenzi, B. M., Lienlaf-Moreno, M., Kandell, W. M., Santiago, D. N., ... & Abate-Daga, D. (2019). An immunoproteomic approach to characterize the CAR interactome and signalosome. Science signaling, 12(568), eaap9777.

Complete CAR data FCM

Fig.7 Flow cytometry analysis of CAR abundance on PSCA-8t28Z (green) and PSCA-8t(+24)28Z (light blue) and GFP (gray)-transduced T cells.

CAR Construction : Ha1-4.117 scFv-28ζ Latest CAR Construction

Fig.7 Flow cytometry analysis of CAR abundance on PSCA-8t28Z (green) and PSCA-8t(+24)28Z (light blue) and GFP (gray)-transduced T cells.

Histograms (left) are representative of 5 independent experiments.

Ramello, M. C., Benzaïd, I., Kuenzi, B. M., Lienlaf-Moreno, M., Kandell, W. M., Santiago, D. N., ... & Abate-Daga, D. (2019). An immunoproteomic approach to characterize the CAR interactome and signalosome. Science signaling, 12(568), eaap9777.

Complete CAR data WB

Fig.8 Western blot analysis of CD3ζ interaction in lysates of T cells
transduced with PSCA-28t28Z, PSCA-8t28BBZ, or GFP and immunoprecipitated for the
CAR scFv.

CAR Construction : Ha1-4.117 scFv-28ζ Latest CAR Construction

Fig.8 Western blot analysis of CD3ζ interaction in lysates of T cells transduced with PSCA-28t28Z, PSCA-8t28BBZ, or GFP and immunoprecipitated for the CAR scFv.

Data are represented as mean±s.d.

Ramello, M. C., Benzaïd, I., Kuenzi, B. M., Lienlaf-Moreno, M., Kandell, W. M., Santiago, D. N., ... & Abate-Daga, D. (2019). An immunoproteomic approach to characterize the CAR interactome and signalosome. Science signaling, 12(568), eaap9777.

Complete CAR data FuncS

Fig.9 CAR signaling upon antigen binding.

CAR Construction : Ha1-4.117 scFv-28ζ Latest CAR Construction

Fig.9 CAR signaling upon antigen binding.

Proteomics data analysis of the 40/751 phospho-proteins that were differentially abundant in CAR-T cells when compared to
background controls.

Ramello, M. C., Benzaïd, I., Kuenzi, B. M., Lienlaf-Moreno, M., Kandell, W. M., Santiago, D. N., ... & Abate-Daga, D. (2019). An immunoproteomic approach to characterize the CAR interactome and signalosome. Science signaling, 12(568), eaap9777.

More Published Data More Published Data

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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.

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