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pCDCAR1 CD20 h(28ζ), T (CAR-T-2-M302-2Z)


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

The vector of anti-CD20 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human CD20. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-CD20 antibody linked to CD28 transmembrane domain/ endodomain and CD3-zeta signaling domains. And the vector product was designed for the treatment of CD20+ malignancies, B-cell lymphoma, mantle cell lymphoma, CD20+ diffuse large cell lymphoma (DLCL), leukemias.

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Details

  • Target
  • CD20
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • CD20+ malignancies; B-cell lymphoma; mantle cell lymphoma; CD20+ diffuse large cell lymphoma (DLCL); leukemias
  • 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-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.
    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
  • 1F5
  • Host
  • Mouse
  • Target Species
  • Human
  • Gene Name
  • membrane spanning 4-domains A1
  • Synonyms
  • B1; S7; Bp35; CD20; CVID5; MS4A2; LEU-16; BA0185

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  • Published Data
WB

Fig.1 Detection of 1F5 scFv-Ig by Western blot analysis.

CAR Construction : Latest CAR Construction

Fig.1 Detection of 1F5 scFv-Ig by Western blot analysis.

1F5 scFv-Ig from COS cell supernatants were detected by probing Western blots using alkaline phosphatase-conjugated GAH after separating these proteins on reducing (A) and nonreducing (B) SDS-PAGE gels. Neg., Supernatants from cells transfected with pCDM8 vector without inserts.

Daming. S., Oliver. W. P., Theta. T., Martha. S., Hayden, J. A. (1999). Characterization of scFv-Ig Constructs Generated from the Anti-CD20 mAb 1F5 Using Linker Peptides of Varying Lengths1. J Immunol 1 June, 162 (11), 6589-6595.

FCM

Fig.2 1F5 scFv-Ig binding demonstrated by flow cytometry.

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Fig.2 1F5 scFv-Ig binding demonstrated by flow cytometry.

A total of 105 Jurkat or Ramos cells were incubated with PBS or 50 μg/ml 1F5 scFv-Ig for 45 min at 4°C followed by washing and incubation with FITC-labeled GAH for 30 min at 4°C.

Daming. S., Oliver. W. P., Theta. T., Martha. S., Hayden, J. A. (1999). Characterization of scFv-Ig Constructs Generated from the Anti-CD20 mAb 1F5 Using Linker Peptides of Varying Lengths1. J Immunol 1 June, 162 (11), 6589-6595.

ELISA

Fig.3 Binding activity of 1F5 scFv-Ig demonstrated by ELISA.

CAR Construction : Latest CAR Construction

Fig.3 Binding activity of 1F5 scFv-Ig demonstrated by ELISA.

A total of 105 Ramos cells were incubated with different concentrations of 1F5 scFv-Ig, followed by incubation with a peroxidase-conjugated GAH second Ab.

Daming. S., Oliver. W. P., Theta. T., Martha. S., Hayden, J. A. (1999). Characterization of scFv-Ig Constructs Generated from the Anti-CD20 mAb 1F5 Using Linker Peptides of Varying Lengths1. J Immunol 1 June, 162 (11), 6589-6595.

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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