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pCDCAR1 PSCA h(ζ) (CAR-YF411)


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 and 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 vector
  • 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 CD21-zeta to transmit a proliferative/survival signal, or all three can be used together.

Target

  • Clone
  • bm2B3
  • Host
  • Mouse
  • 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.

CAR Construction : bm2B3 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.

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.2 PSCA CARs effectively treat established tumors in NSG mice

CAR Construction : bm2B3 scFv-CD28-4-1BB-CD3ζ Latest CAR Construction

Fig.2 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 IHC

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

CAR Construction : bm2B3 scFv-CD28-4-1BB-CD3ζ Latest CAR Construction

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

Immunohistochemical analysis of FAP in PSCA-28Z CAR-treated tumor and control GFP-treated tumor.

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.4 iMC Provides Rimiducid-Dependent Costimulation for CAR-T Cells

CAR Construction : bm2B3 scFv-28ζ Latest CAR Construction

Fig.4 iMC Provides Rimiducid-Dependent Costimulation for CAR-T Cells

T cells transduced (n = 3) with iMC plus PSCA.z were cultured in media without exogenous IL-2 and stimulated with 10 nM Rim on a weekly basis, where indicated.

Foster, A. E., Mahendravada, A., Shinners, N. P., Chang, W. C., Crisostomo, J., Lu, A., ... & Spencer, D. M. (2017). Regulated expansion and survival of chimeric antigen receptor-modified T cells using small molecule-dependent inducible MyD88/CD40. Molecular therapy, 25(9), 2176-2188.

Complete CAR data ELISA

Fig.5 T cells were transduced with EGFPluc or with both EGFPluc and iMC-PSCA.z-encoding-retrovirus and subsequently cultured with HPAC-RFP tumor cells
at a 1:10 effector-to-tumor (E:T) cell ratio for 48 hr with variable Rim concentrations (0–250 nM) (n = 3).

CAR Construction : bm2B3 scFv-28ζ Latest CAR Construction

Fig.5 T cells were transduced with EGFPluc or with both EGFPluc and iMC-PSCA.z-encoding-retrovirus and subsequently cultured with HPAC-RFP tumor cells at a 1:10 effector-to-tumor (E:T) cell ratio for 48 hr with variable Rim concentrations (0–250 nM) (n = 3).

Data are represented as mean±s.d.

Foster, A. E., Mahendravada, A., Shinners, N. P., Chang, W. C., Crisostomo, J., Lu, A., ... & Spencer, D. M. (2017). Regulated expansion and survival of chimeric antigen receptor-modified T cells using small molecule-dependent inducible MyD88/CD40. Molecular therapy, 25(9), 2176-2188.

Complete CAR data FuncS

Fig.6 EGFPluc and iMC-PSCA.z/EGFPluc-modified
T cells were cultured with HPAC-RFP tumor cells at an E:T ratio of 1:20 and imaged over 7 days (n = 2). T cells transduced with iMC-PSCA.z/EGFPluc were stimulated with or
without 2 nM Rim on day 0 of culture initiation.

CAR Construction : bm2B3 scFv-28ζ Latest CAR Construction

Fig.6 EGFPluc and iMC-PSCA.z/EGFPluc-modified T cells were cultured with HPAC-RFP tumor cells at an E:T ratio of 1:20 and imaged over 7 days (n = 2). T cells transduced with iMC-PSCA.z/EGFPluc were stimulated with or without 2 nM Rim on day 0 of culture initiation.

Validation of a Unified Vector Encoding iMC and PSCA.z CAR

Foster, A. E., Mahendravada, A., Shinners, N. P., Chang, W. C., Crisostomo, J., Lu, A., ... & Spencer, D. M. (2017). Regulated expansion and survival of chimeric antigen receptor-modified T cells using small molecule-dependent inducible MyD88/CD40. Molecular therapy, 25(9), 2176-2188.

Complete CAR data BLI

Fig.7 Tumor BLI was assessed weekly (dotted lines indicate individual animals and full circles show group average).

CAR Construction : bm2B3 scFv-28ζ Latest CAR Construction

Fig.7 Tumor BLI was assessed weekly (dotted lines indicate individual animals and full circles show group average).

These data are representative of three independent experiments.

Foster, A. E., Mahendravada, A., Shinners, N. P., Chang, W. C., Crisostomo, J., Lu, A., ... & Spencer, D. M. (2017). Regulated expansion and survival of chimeric antigen receptor-modified T cells using small molecule-dependent inducible MyD88/CD40. Molecular therapy, 25(9), 2176-2188.

Complete CAR data TCA

Fig.8 T cells were cotransduced with iMC-PSCA.z and EGFPluc and injected i.v. into tumor-free NSG mice. Mice (n = 5 per group) were either left untreated or treated with a single i.p. injection of Rim at 5 mg/kg 24 hr post-T cell injection.

CAR Construction : bm2B3 scFv-28ζ Latest CAR Construction

Fig.8 T cells were cotransduced with iMC-PSCA.z and EGFPluc and injected i.v. into tumor-free NSG mice. Mice (n = 5 per group) were either left untreated or treated with a single i.p. injection of Rim at 5 mg/kg 24 hr post-T cell injection.

Radiance was calculated from in vivo BLI measurements on days +1, +8, and +30 post-T cell injection (LoD or limit of detection for luminescence).

Foster, A. E., Mahendravada, A., Shinners, N. P., Chang, W. C., Crisostomo, J., Lu, A., ... & Spencer, D. M. (2017). Regulated expansion and survival of chimeric antigen receptor-modified T cells using small molecule-dependent inducible MyD88/CD40. Molecular therapy, 25(9), 2176-2188.

Complete CAR data BI

Fig.9 NSG mice (n = 5 per group) were engrafted with HPAC-EGFPluc and treated with 1.0 106 iMC-PSCA.z-modified T cells or NT cells after 7 days.

CAR Construction : bm2B3 scFv-28ζ Latest CAR Construction

Fig.9 NSG mice (n = 5 per group) were engrafted with HPAC-EGFPluc and treated with 1.0 106 iMC-PSCA.z-modified T cells or NT cells after 7 days.

Mice subsequently received weekly i.p. vehicle (Veh) or Rim injections of 5 mg/kg

Foster, A. E., Mahendravada, A., Shinners, N. P., Chang, W. C., Crisostomo, J., Lu, A., ... & Spencer, D. M. (2017). Regulated expansion and survival of chimeric antigen receptor-modified T cells using small molecule-dependent inducible MyD88/CD40. Molecular therapy, 25(9), 2176-2188.

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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