Anti-CD117 (CK6) h(41BB-CD3ζ) CAR, pMMLV vector

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

Recombinant Moloney murine leukemia virus (MMLV) retroviral vectors serve as efficient viral vector tools for introducing genes permanently into a wide variety of dividing cells, and provide an alternative to modify primary T cells. Creative biolabs has developed retroviral CAR vector pMMLV CD117 (CK6) h(BBζ), which is constructed for the engineering of T cells to target human CD117. The T cells are genetically modified through transduction with a retroviral vector expressing scFv of anti-CD117 antibody linked to CD137 (4-1BB) and CD3-zeta signaling domains. And the vector product was designed for the treatment of Gastrointestinal stromal tumors, Mast cell disease, Acute myelogenous lukemia, Piebaldism.

Details

Target
CD117

Targeting Cell Type
T cell

Targeting Diseases
Gastrointestinal stromal tumors, Mast cell disease, Acute myelogenous lukemia, Piebaldism

Generation
Second

Vector Name
pMMLV

Vector Length
~6kb

Vector Type
Recombinant Moloney murine leukemia virus (MMLV) retroviral vector

Receptor Construction
scFv-41BB-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 antigennonspecific 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 4-1BB and OX40 can be used with CD3-zeta to transmit a proliferative/survival signal, or all three can be used together.

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Published Data

CAR scFv data ELISA

Fig.1 CK6 binding to immobilized human c-Kit protein (EC50 = 0.06 nM) in ELISA.

CAR Construction :

Fig.1 CK6 binding to immobilized human c-Kit protein (EC50 = 0.06 nM) in ELISA.

One representative experiment is shown with EC50.

Lebron, M. B., Brennan, L., Damoci, C. B., Prewett, M. C., O’Mahony, M., Duignan, I. J., ... & Kotanides, H. (2014). A human monoclonal antibody targeting the stem cell factor receptor (c-Kit) blocks tumor cell signaling and inhibits tumor growth. Cancer biology & therapy, 15(9), 1208-1218.

CAR scFv data FCM

Fig 2. Flow cytometry staining analysis of CK6 binding to c-Kit receptor expressing SCLC tumor cell lines.

CAR Construction :

Fig 2. Flow cytometry staining analysis of CK6 binding to c-Kit receptor expressing SCLC tumor cell lines.

Histograms indicate fluorescence signal of unstained cells (solid gray fill) and cells stained with either CK6 (black solid line) or human IgG (black dashed line).

CAR scFv data FCM

Fig 3. Flow cytometry staining analysis of CK7 binding to c-Kit receptor expressing melanoma tumor cell lines.

CAR Construction :

Fig 3. Flow cytometry staining analysis of CK7 binding to c-Kit receptor expressing melanoma tumor cell lines.

Histograms indicate fluorescence signal of unstained cells (solid gray fill) and cells stained with either CK6 (black solid line) or human IgG (black dashed line).

CAR scFv data FCM

Fig 4. Flow cytometry staining analysis of CK8 binding to c-Kit receptor expressing leukemia cell lines.

CAR Construction :

Fig 4. Flow cytometry staining analysis of CK8 binding to c-Kit receptor expressing leukemia cell lines.

Histograms indicate fluorescence signal of unstained cells (solid gray fill) and cells stained with either CK6 (black solid line) or human IgG (black dashed line).

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.

Anti-CD117 (CK6) h(41BB-CD3ζ) CAR, pMMLV (CAR-MV-01LX453)

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