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pCDCAR1 Lewis Y h(28ζ) (CAR-ZP8394)

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

The vector of anti-Lewis Y chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target Human Lewis Y. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-Lewis Y antibody linked to CD28 and CD3ζ signaling domains. And the vector product was designed for the treatment of gastric cancer.

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Details

  • Target
  • Lewis Y
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • gastric 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 costimulatory 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
  • BR55-2
  • Host
  • Mouse
  • Target Species
  • Human
  • Gene Name
  • Lewis Y
  • Synonyms
  • Lewis Y

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  • Published Data
CAR scFv data FCM

Fig.2 Flow cytometric analysis of binding activity of mAbPand mAbMBR55-2 to the FcRI receptor.

CAR Construction : Latest CAR Construction

Fig.2 Flow cytometric analysis of binding activity of mAbPand mAbMBR55-2 to the FcRI receptor.

U937 cells treated with IFN to stimulate Fc receptorexpression were incubated with mAbPor mAbMBR55-2 or CD64-specific mAb m22-FITC.

Brodzik, R., Glogowska, M., Bandurska, K., Okulicz, M., Deka, D., Ko, K., ... & Koprowski, H. (2006). Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells. Proceedings of the National Academy of Sciences, 103(23), 8804-8809.

CAR scFv data ELISA

Fig.3 ELISA plates were coated overnight with the peptides, and reactivity of serial dilutions of the mAbs was detected.

CAR Construction : Latest CAR Construction

Fig.3 ELISA plates were coated overnight with the peptides, and reactivity of serial dilutions of the mAbs was detected.

Plates were then washed and incubated with post-immune serum from a patient immunized with conjugated P10s.

Nounamo, B., Jousheghany, F., Siegel, E. R., Post, S. R., Kelly, T., Ferrone, S., ... & Monzavi-Karbassi, B. (2023). VT68. 2: An Antibody to Chondroitin Sulfate Proteoglycan 4 (CSPG4) Displays Reactivity against a Tumor-Associated Carbohydrate Antigen. International Journal of Molecular Sciences, 24(3), 2506.

CAR scFv data FCM

Fig.1 MDA-MB-231 cells were preincubated with Caspase-3 Inhibitor Z-DEVD-FMK or DMSO (control) for 2 hours and then pre- and post-immune plasma were added and incubation continued overnight.

CAR Construction : Latest CAR Construction

Fig.1 MDA-MB-231 cells were preincubated with Caspase-3 Inhibitor Z-DEVD-FMK or DMSO (control) for 2 hours and then pre- and post-immune plasma were added and incubation continued overnight.

Cells then were harvested and stained with annexin V-FITC (FL1) and propidium iodide (FL3) using a live/dead assay kit.

Hutchins, L. F., Makhoul, I., Emanuel, P. D., Pennisi, A., Siegel, E. R., Jousheghany, F., ... & Kieber-Emmons, T. (2017). Targeting tumor-associated carbohydrate antigens: a phase I study of a carbohydrate mimetic-peptide vaccine in stage IV breast cancer subjects. Oncotarget, 8(58), 99161.

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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