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pCDCAR1 ROR1 h(28ζ) (CAR-YF113)


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

The vector of anti-ROR1 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human ROR1. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-ROR1 antibody linked to CD28 and CD3ζ signaling domains. And the vector product was designed for the treatment of ROR1 -positive leukemia, mantle cell lymphoma, breast- cancer, lung cancer.

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Details

  • Target
  • ROR1
  • Targeting Cell Type
  • T cell
  • Targeting Diseases
  • Ror1 -Positive Leukemia, Mantle Cell Lymphoma, Breast- Cancer, Lung 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 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 cell 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
  • R11
  • Host
  • Rabbit
  • Target Species
  • Human
  • Gene Name
  • Receptor tyrosine kinase like orphan receptor 1
  • Synonyms
  • ROR1; NTRKR1; dJ537F10.1

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  • Published Data
CAR scFv data FCM

Fig.1 The epitopes of IgG1 R11, R12, and Y31 were mapped with five immobilized Fc fusion proteins that consisted of just one or two extracellular domains of human ROR1.

CAR Construction : Latest CAR Construction

Fig.1 The epitopes of IgG1 R11, R12, and Y31 were mapped with five immobilized Fc fusion proteins that consisted of just one or two extracellular domains of human ROR1.

Columns indicate mean values and error bars indicate standard deviation values of side-by-side triplicates.

Yang, J., Baskar, S., Kwong, K. Y., Kennedy, M. G., Wiestner, A., & Rader, C. (2011). Therapeutic potential and challenges of targeting receptor tyrosine kinase ROR1 with monoclonal antibodies in B-cell malignancies. PloS one, 6(6), e21018.

CAR scFv data SPR

Fig.2 Epitope mapping studies by surface plasmon resonance.

CAR Construction : Latest CAR Construction

Fig.2 Epitope mapping studies by surface plasmon resonance.

Shown are Biacore 6100 sensorgrams obtained for the binding of IgG1 R11, R12, and Y31 to immobilized Fc-hROR1.

Yang, J., Baskar, S., Kwong, K. Y., Kennedy, M. G., Wiestner, A., & Rader, C. (2011). Therapeutic potential and challenges of targeting receptor tyrosine kinase ROR1 with monoclonal antibodies in B-cell malignancies. PloS one, 6(6), e21018.

CAR scFv data SPR

Fig.3 Affinity measurements of R12 by surface plasmon resonance.

CAR Construction : Latest CAR Construction

Fig.3 Affinity measurements of R12 by surface plasmon resonance.

The mAbs were injected at five or six different concentrations (shown in different colors) ranging from 1.5 to 100 nM. Each concentration was tested in duplicates.

Yang, J., Baskar, S., Kwong, K. Y., Kennedy, M. G., Wiestner, A., & Rader, C. (2011). Therapeutic potential and challenges of targeting receptor tyrosine kinase ROR1 with monoclonal antibodies in B-cell malignancies. PloS one, 6(6), e21018.

CAR scFv data FCM

Fig.4 Flow cytometry was used to analyze the binding of IgG1 R11 (green line; 5 mg/mL), R12 (blue line; 1 mg/mL), and Y31 (red line; 5 mg/mL) to (A) the surface of JeKo-1 (top) and HBL-2 cells (bottom).

CAR Construction : Latest CAR Construction

Fig.4 Flow cytometry was used to analyze the binding of IgG1 R11 (green line; 5 mg/mL), R12 (blue line; 1 mg/mL), and Y31 (red line; 5 mg/mL) to (A) the surface of JeKo-1 (top) and HBL-2 cells (bottom).

Biotinylated IgG1 in combination with FITC-CD19/APC-CD5 were detected with PE-streptavidin. The y axis depicts the number of events, the x axis the fluorescence intensity.

Yang, J., Baskar, S., Kwong, K. Y., Kennedy, M. G., Wiestner, A., & Rader, C. (2011). Therapeutic potential and challenges of targeting receptor tyrosine kinase ROR1 with monoclonal antibodies in B-cell malignancies. PloS one, 6(6), e21018.

CAR scFv data FCM

Fig.5 PI staining followed by flow cytometry was used to analyze the CDC potency of IgG1 R11, R12, and Y31 in comparison to IgG1 P14 (negative control), and rituximab.

CAR Construction : Latest CAR Construction

Fig.5 PI staining followed by flow cytometry was used to analyze the CDC potency of IgG1 R11, R12, and Y31 in comparison to IgG1 P14 (negative control), and rituximab.

The percentage of dead cells (PI-positive) after RTX-mediated CDC was 70.7% (JeKo-1), 54.4% (HBL-2), and 12.7% (CLL).

Yang, J., Baskar, S., Kwong, K. Y., Kennedy, M. G., Wiestner, A., & Rader, C. (2011). Therapeutic potential and challenges of targeting receptor tyrosine kinase ROR1 with monoclonal antibodies in B-cell malignancies. PloS one, 6(6), e21018.

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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