Description
Pre-made Lentivirus express the CAR construct of “3xFLAG-anti- HLA-A2-ScFV (VL-VH)-CD8 hinge-4-1BB-CD3ζ”. It contains the GFP-Puromycin dual marker, allowing the selection of the transduced cells via antibiotic killing, or via GFP positive cell sorting.
This construct, contains the N-term 3xFLAG tagged ” HLA-A2-scFv [VH-(G4S)x4-VL]” recognition domain, linked to 4-1BB stimulatory domain, expressed under the enhance EF1a promoter. The lentivector’s core structure is represented by the scheme below.
HLA-A2 is a major histocompatibility complex (MHC) class I molecule involved in presenting antigens to cytotoxic T cells. HLA-A2 is not a traditional target for CAR-T therapy, as CARs typically recognize cell surface antigens rather than MHC molecules. However, the context in which HLA-A2 is mentioned might involve its role in antigen presentation. CARs anti HLA is used to engineering human regulatory T cells (Tregs), and is a promising strategy to induce immune tolerance in transplants. This CAR construct assembled the coding sequence of the human monoclonal anti-HLA-A2 antibody (clone BB7.2 and SN607DB hybridoma). (see Reference1, Reference2).
see details in Product Manual.
4-1BB (CD137) is a surface glycoprotein present on activated T Lymphocytes, CD4+ and CD8+ T cells. It binds to its ligand expressed on antigen-presenting cells (macrophages and activated B cells). It promotes T-cell antitumor activity. 4-1BB is one of the most widely studied co-stimulatory domain used in CAR design.
CD3ζ (T-cell receptor zeta) is expressed by T cells and NK cells. It together with T-cell receptor and CD3γ, δ, ε chain, forms the TCR-CD3 complex. CD3-zeta is the most commonly used activation component of CARs. It transmits an activation signal to the T cell after the antigen is bound. It can be coupled with additional co-stimulatory signaling for the complete activation.
Negative Controls: To validate the specificity of the targeted CAR-T killing, you can use the negative control CAR lentivirus: the CAR-ctr7 that has the same lentivector backbone but removed the recognition domain.
Amount: 200 ul/vial at titer of 1×108 IFU/ml, concentrated lentivirus provided in PBS solution, premixed with 10x polybrene (60ug/ml), premade, ready to ship in dry-ice package.
CAT#: LVP1667
