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pCDCAR1 CD22 h(28ζ) (CAR-LC061)


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

The vector of anti-CD22 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human CD22. The T cells are genetically modified through transduction with a retroviral vector expressing scFv of anti-CD22 antibody linked to CD28 and CD3ζ signaling domains. And the vector product was designed for the treatment of acute lymphoblastic leukemia.

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Details

  • Target
  • CD22
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Acute lymphoblastic leukemia
  • Generation
  • Second
  • Vector Name
  • pCDCAR1
  • Vector Length
  • ~8kb
  • Vector Type
  • Retroviral
  • 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
  • HA22
  • Host
  • Mouse
  • Target Species
  • Human
  • Gene Name
  • CD22
  • Synonyms
  • CD22;CD22; CD22 Molecule; CD22 Molecule; CD22 Antigen; Sialic Acid-Binding Ig-Like Lectin 2; B-Lymphocyte Cell Adhesion Molecule; T-Cell Surface Antigen Leu-14; SIGLEC-2;

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  • Published Data
Complete CAR data FuncS

Fig.1 Expression on transduced T cells of CD22 CARs in the second and third-generation constructs.

CAR Construction : HA22-CD28-CD3ζ, HA22-CD28-41BB-CD3ζ Latest CAR Construction

Fig.1 Expression on transduced T cells of CD22 CARs in the second and third-generation constructs.

CD22 CARs were detected by CD3-APC (y-axis) and CD22-Fc followed by an anti-IgG-Fc-FITC stain. Percent transduction is noted in the top-right quadrant of each plot.

Haso, W., Lee, D. W., Shah, N. N., Stetler-Stevenson, M., Yuan, C. M., Pastan, I. H., ... & Orentas, R. J. (2013). Anti-CD22–chimeric antigen receptors targeting B-cell precursor acute lymphoblastic leukemia. Blood, The Journal of the American Society of Hematology, 121(7), 1165-1174.

Complete CAR data ELISA

Fig.2 HA22 and BL22-derived CD22 CARs mediate lytic activity.

CAR Construction : HA22-CD28-41BB-CD3ζ Latest CAR Construction

Fig.2 HA22 and BL22-derived CD22 CARs mediate lytic activity.

The lytic activity HA22 and BL22-derived CD22 CARs was compared against ALL cell lines at the indicated E:T ratios in a 4-hour 51Cr-release assay.

Haso, W., Lee, D. W., Shah, N. N., Stetler-Stevenson, M., Yuan, C. M., Pastan, I. H., ... & Orentas, R. J. (2013). Anti-CD22–chimeric antigen receptors targeting B-cell precursor acute lymphoblastic leukemia. Blood, The Journal of the American Society of Hematology, 121(7), 1165-1174.

CAR scFv data FCM

Fig.3 Interaction between CD22 and HEK 293T cell surface-displayed scFv

CAR Construction : Latest CAR Construction

Fig.3 Interaction between CD22 and HEK 293T cell surface-displayed scFv

Four individual clones (PT.1, PT.2, PT.3, and PT.4) of scFvs with the same mutation (PT) isolated from a single-pass enrichment were tested for the scFv expression (scFv) on HEK 293T cells with anti-myc mAb detected by FITC and for the binding to the antigen with CD22-Fc (CD22) detected by PE.

Ho, M., Nagata, S., & Pastan, I. (2006). Isolation of anti-CD22 Fv with high affinity by Fv display on human cells. Proceedings of the National Academy of Sciences, 103(25), 9637-9642.

CAR scFv data FCM

Fig.4 Equilibrium binding titration curves to determine dissociation constants, KD.

CAR Construction : Latest CAR Construction

Fig.4 Equilibrium binding titration curves to determine dissociation constants, KD.

MFI (%) of PE is plotted versus the various concentrations of biotinylated CD22-Fc used to label surface-displayed BL22 (WT, KD = 5.8 nM, Bmax = 453), HA22 (KD = 2.5 nM, Bmax = 293), or mutant PT (KD = 1.2 nM, Bmax = 275) antibody.

Ho, M., Nagata, S., & Pastan, I. (2006). Isolation of anti-CD22 Fv with high affinity by Fv display on human cells. Proceedings of the National Academy of Sciences, 103(25), 9637-9642.

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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