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pCDCAR1 CAIX h(28ζ) (CAR-MZ202)


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

The vector of anti-Carbonic anhydrase IX (CAIX) chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human Carbonic anhydrase IX (CAIX). The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-Carbonic anhydrase IX (CAIX) antibody linked to CD28 and CD3ζ signaling domains. And the vector product was designed for the treatment of Clear cell renal cell carcinoma (ccRCC).

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Details

  • Target
  • Carbonic anhydrase IX (CA9)
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Clear cell renal cell carcinoma (ccRCC)
  • Generation
  • Second
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Lentiviral
  • 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 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

  • Clone
  • G36
  • Host
  • Mouse
  • Target Species
  • Human
  • Gene Name
  • CA9
  • Synonyms
  • CA9; CAIX; MN; carbonic anhydrase 9

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  • Published Data
FACS

Fig.1 Fluorescence-activated cell sorting analysis.

CAR Construction : G36 scFv-CD28-TCRζ (CD28 CAR) Latest CAR Construction

Fig.1 Fluorescence-activated cell sorting analysis.

Reporter gene ZsGreen was used to quantitate primary T cell transduction efficiency by the lentiviral CAR constructs. In addition, anti-CAIX scFv CARs were stained with CAIX-Fc fusion protein and C9-tag (TETSQVAPA) was stained with 1D4 antibody. Untransduced activated T-cells, LAK only were served as unstained cell control (i) or stained with second antibody ((ii) PE-anti-human IgG and (iii) APC-anti-mouse IgG) were used as staining controls.

Lo, A. S., Xu, C., Murakami, A., & Marasco, W. A. (2014). Regression of established renal cell carcinoma in nude mice using lentivirus-transduced human T cells expressing a human anti-CAIX chimeric antigen receptor. Molecular therapy oncolytics, 1, 14003.

WB

Fig.2 Western blot analysis.

CAR Construction : G36 scFv-CD28-TCRζ (CD28 CAR) Latest CAR Construction

Fig.2 Western blot analysis.

Molecular sizes of monomer/dimer structures of anti-CAIX (clone G36) CD28 and anti-CCR5 (clone A8) CD28 CARs, as well as endogenous TCRζ chain of untransduced T cells were indicated.

Lo, A. S., Xu, C., Murakami, A., & Marasco, W. A. (2014). Regression of established renal cell carcinoma in nude mice using lentivirus-transduced human T cells expressing a human anti-CAIX chimeric antigen receptor. Molecular therapy oncolytics, 1, 14003.

FuncS

Fig.3 Functional studies.

CAR Construction : G36 scFv-CD28-TCRζ (CD28 CAR) Latest CAR Construction

Fig.3 Functional studies.

Cytokine secretion. Anti-CAIX CART, irrelevant CART, or activated control T cells (LAK) were cocultivated overnight with kidney cancer cell lines sk-rc-52 (CAIX+) and sk-rc-59 (CAIX−) for cytokine production. One representative out of two to three results is shown.

Lo, A. S., Xu, C., Murakami, A., & Marasco, W. A. (2014). Regression of established renal cell carcinoma in nude mice using lentivirus-transduced human T cells expressing a human anti-CAIX chimeric antigen receptor. Molecular therapy oncolytics, 1, 14003.

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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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