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pCDCAR1 CAIX h(28γ), T (CAR-T-2-M305-2G)


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

The vector of anti-CAIX chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human CAIX. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-CAIX antibody linked to CD28 transmembrane domain/ endodomain and FcεRIγ signaling domains. And the vector product was designed for the treatment of renal cell carcinoma (RCC).

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Details

  • Target
  • Carbonic anhydrase IX (CA9)
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Renal cell carcinoma (RCC)
  • Generation
  • Second
  • Vector Name
  • pCDCAR1
  • Vector Length
  • 8kb
  • Vector Type
  • Lentiviral
  • Receptor Construction
  • scFv-CD28-FcεRIγ
  • Discription of Signaling Cassetes
  • CD28
    CD28 (Cluster of Differentiation 28) is one of the proteins expressed on T cells that provide co-stimulatorysignals 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.
    FcεRIγ
    The high-affinity IgE receptor, also known as FcεRI, or Fc epsilon RI, is the high-affinity receptor for the Fc region of immunoglobulin E (IgE). FcεRI is a key molecule involved in allergic reactions. It is a tetramer composed of 1alpha, 1 beta, and 2 gamma chains. The gamma chains are also subunits of other Fc receptors. The scFv-based CARs engineered to contain a signaling domain from FcεRIγ have been shown to deliver a potent signal for T cell activation and function.

Target

  • Clone
  • G250
  • 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 : G250 scFv-41BB-CD28ζ Latest CAR Construction

Fig.1 Fluorescence-activated cell sorting analysis.

CAR-positive rate of lentivirus-transduced NK92 cells. NK92 cells were transduced with empty lentivirus vector (Ctrl-NK92) or lentivirus containing the CAIX-specific CAR encoding sequence (CAR-NK92) and were selected with puromycin.

Zhang, Q., Xu, J., Ding, J., Liu, H., Li, H., Li, H., Lu, M., Miao, Y., Wang, Z., Fu, Q., & Zheng, J. (2018). Bortezomib improves adoptive carbonic anhydrase IX‑specific chimeric antigen receptor‑modified NK92 cell therapy in mouse models of human renal cell carcinoma. Oncology reports, 40(6), 3714–3724.

WB

Fig.2 Western blot analysis.

CAR Construction : G250 scFv-41BB-CD28ζ Latest CAR Construction

Fig.2 Western blot analysis.

Western blot analysis of the CAR expression in NK92 cells with a monoclonal anti-human CD3ζ antibody. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was also detected as an internal control.

Zhang, Q., Xu, J., Ding, J., Liu, H., Li, H., Li, H., Lu, M., Miao, Y., Wang, Z., Fu, Q., & Zheng, J. (2018). Bortezomib improves adoptive carbonic anhydrase IX‑specific chimeric antigen receptor‑modified NK92 cell therapy in mouse models of human renal cell carcinoma. Oncology reports, 40(6), 3714–3724.

FuncS

Fig.3 Functional studies.

CAR Construction : G250 scFv-41BB-CD28ζ Latest CAR Construction

Fig.3 Functional studies.

Specific cytokine release of CAIX-specific CAR-NK92 cells against CAIX+ cells.

Zhang, Q., Xu, J., Ding, J., Liu, H., Li, H., Li, H., Lu, M., Miao, Y., Wang, Z., Fu, Q., & Zheng, J. (2018). Bortezomib improves adoptive carbonic anhydrase IX‑specific chimeric antigen receptor‑modified NK92 cell therapy in mouse models of human renal cell carcinoma. Oncology reports, 40(6), 3714–3724.

Cyt

Fig.4 Cytotoxicity assay.

CAR Construction : G250 scFv-41BB-CD28ζ Latest CAR Construction

Fig.4 Cytotoxicity assay.

Specific cytotoxicity exhibited by CAIX-specific CAR-NK92 cells against CAIX+ target cells.

Zhang, Q., Xu, J., Ding, J., Liu, H., Li, H., Li, H., Lu, M., Miao, Y., Wang, Z., Fu, Q., & Zheng, J. (2018). Bortezomib improves adoptive carbonic anhydrase IX‑specific chimeric antigen receptor‑modified NK92 cell therapy in mouse models of human renal cell carcinoma. Oncology reports, 40(6), 3714–3724.

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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