Anti-RoR1 (R11) h(41BB-CD3ζ) CAR, pCDCAR1 vector

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

Details

Target
RoR1

Targeting Cell Type
T cell

Targeting Diseases
Lymphoma, breast cancer, ovarian cancer, lung cancer, pancreatic cancer and prostate cancer

Generation
Second

Vector Name
pCDCAR1

Vector Length
~8kb

Vector Type
Lentiviral vector

Receptor Construction
scFv-4-1BB-CD3ζ

Discription of Signaling Cassetes
41BB
CD137 (also known as 4-1BB) is a surface co-stimulatory glycoprotein originally described as present on activated T lymphocytes, which belongs to the tumor necrosis factor (TNF) receptor superfamily. It is expressed mainly on activated CD4+ and CD8+ T cells, and binds to a high-affinity ligand (4-1BBL) expressed on several antigen-presenting cells such as macrophages and activated B cells. On the basis of preclinical observation, this molecule can promote the persistence of antigen-specific and antigen-nonspecific chimeric antigen receptor T-cells to significantly increases antitumor activity.
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

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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 :

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 :

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.

CAR scFv data SPR

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

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.

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 :

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.

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 :

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).

More Published Data More Published Data

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.