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Anti-EphA2 (B208)-FcRɣ+PI3K CAR THP1 (CARTHP-W0666)

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

The anti-EphA2 chimeric antigen receptor (CAR) THP1 cell line is a stable cell line developed using anti-EphA2 CAR lentivirus transfection. This transfected CAR lentiviral vector is constructed to express a scfv of anti-EphA2 antibody (B208) linked to FcRɣ+PI3K signaling domains.The anti-EphA2 CAR engineered THP1 cells can be used to be differentiated into macrophage cells for in vitro or in vivo phagocytosis studies. This CAR-THP1 cell line product can be used for the researches of GBM, glioma.

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  • Marker:
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Details

  • Target
  • EphA2
  • Cell Type
  • THP1
  • Cell Background
  • THP-1 is a human leukemia monocytic cell line, which has been extensively used to study monocyte/macrophage functions, mechanisms, signaling pathways, and nutrient and drug transport. This cell line has become a common model to estimate modulation of monocyte and macrophage activities. CAR engineered THP1 cells can be differentiated into macrophages using PMA (Phorbol 12-myristate 13-acetate). Once differentiated (M0 macrophages), they were incubated with IL-4 and IL-13 in order to obtain M2 polarized macrophages or with IFN-gamma and LPS for classical macrophage activation (M1).
  • Targeting Diseases
  • GBM, glioma
  • CAR Expression
  • Positive
  • CAR Positive Rate
  • >95% for stable cell pool (please submit an inquiry if single clonal cell line is needed.)
  • Cell Viability
  • >90%
  • Mycoplasma Testing
  • Negative
  • Handling Notes
  • Frozen cells should be thawed immediately upon receipt and grown according to handling procedure to ensure cell viability and proper assay performance.
    Note: Do not freeze the cells upon receipt as it may result in irreversible damage to the cell line.
    Disclaimer: We cannot guarantee cell viability if the cells are not thawed immediately upon receipt and grown according to handling procedure.
  • Safety Consideration
  • Biosafety Level 1
  • Restriction
  • Research use only
  • Size
  • 2 million cells*2 vials
  • Storage
  • Frozen cells should be stored in a liquid nitrogen tank (-150°C~-190°C) for long term.

CAR Components

  • Promoter
  • EF1a
  • scFv
  • B208
  • Hinge
  • CD8
  • TM
  • CD8
  • ICD
  • FcRɣ+PI3K
  • Marker
  • GFP or N/A
  • Receptor Construction
  • scFv-hinge(CD8)-TM(CD8)-ICD(FcRɣ+PI3K)
  • Discription of Signaling Cassetes
  • FcRɣ
    FcRɣ is used as the CAR intracellular signaling domain. As a phagocytosis receptor, FcRɣ has cytosolic Immunoreceptor Tyrosine-based Activation Motifs (ITAMs) that are phosphorylated by Src family kinases. Based on this mechanism, FcRɣ signaling can induce macrophage phagocytosis which has been demonstrated in the published article.
    Fc gamma receptors belong to the group of non-catalytic tyrosine-phosphorylated receptors which share a similar signalling pathway involving phosphorylation of tyrosine residues. The receptors generate signals within their cells through an important activation motif known as an immunoreceptor tyrosine-based activation motif (ITAM). An ITAM is present in the intracellular tail of FcγRIIA, and its phosphorylation induces phagocytosis in macrophages. FcγRI and FcγRIIIA do not have an ITAM but can transmit an activating signal to their phagocytes by interacting with another protein that does. This adaptor protein is called the Fcγ subunit and, like FcγRIIA, contains the two YXXL sequences that are characteristic of an ITAM.

    PI3K
    Previous work demonstrated that PI3K signaling is important for internalization of large targets. To increase PI3K recruitment to the CAR-P, we fused the portion of the CD19 cytoplasmic domain (amino acids 500 to 534) that recruits the p85 subunit of PI3K to the CAR. A CAR-P containing the p85 recruitment motif alone (CAR-P-PI3K) was able to induce some whole cell engulfment. Assembling an array of motifs designed to recruit distinct phagocytic effectors can increase CAR-P activity.

Target

  • Target
  • EphA2
  • Target Species
  • Human
  • Clone
  • B208
  • Host
  • Mouse
  • Gene Name
  • EPH receptor A2
  • Introduction
  • This gene belongs to the ephrin receptor subfamily of the protein-tyrosine kinase family. EPH and EPH-related receptors have been implicated in mediating developmental events, particularly in the nervous system. Receptors in the EPH subfamily typically have a single kinase domain and an extracellular region containing a Cys-rich domain and 2 fibronectin type III repeats. The ephrin receptors are divided into 2 groups based on the similarity of their extracellular domain sequences and their affinities for binding ephrin-A and ephrin-B ligands. This gene encodes a protein that binds ephrin-A ligands. Mutations in this gene are the cause of certain genetically-related cataract disorders.
  • Alternative Names
  • ECK; CTPA; ARCC2; CTPP1; CTRCT6; EPHA2; EPH receptor A2; Erythropoetin producing hepatocellular carcinoma A2

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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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