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The vector of anti-CD20 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target human CD20. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-CD20 antibody linked to CD28 transmembrane domain/ endodomain and CD137 (4-1BB), CD3-zeta signaling domains. And the vector product was designed for the treatment of CD20+ malignancies , B-cell lymphoma, mantle cell lymphoma, CD20+ diffuse large cell lymphoma (DLCL), leukemias.
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CAR Construction :
Fig.1 Detection of 1F5 scFv-Ig by Western blot analysis. 1F5 scFv-Ig from COS cell supernatants were detected by probing Western blots using alkaline phosphatase-conjugated GAH after separating these proteins on reducing (A) and nonreducing (B) SDS-PAGE gels. Neg., Supernatants from cells transfected with pCDM8 vector without inserts. Daming. S., Oliver. W. P., Theta. T., Martha. S., Hayden, J. A. (1999). Characterization of scFv-Ig Constructs Generated from the Anti-CD20 mAb 1F5 Using Linker Peptides of Varying Lengths1. J Immunol 1 June, 162 (11), 6589-6595. |
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CAR Construction :
Fig.2 1F5 scFv-Ig binding demonstrated by flow cytometry. A total of 105 Jurkat or Ramos cells were incubated with PBS or 50 μg/ml 1F5 scFv-Ig for 45 min at 4°C followed by washing and incubation with FITC-labeled GAH for 30 min at 4°C. Daming. S., Oliver. W. P., Theta. T., Martha. S., Hayden, J. A. (1999). Characterization of scFv-Ig Constructs Generated from the Anti-CD20 mAb 1F5 Using Linker Peptides of Varying Lengths1. J Immunol 1 June, 162 (11), 6589-6595. |
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CAR Construction :
Fig.3 Binding activity of 1F5 scFv-Ig demonstrated by ELISA. A total of 105 Ramos cells were incubated with different concentrations of 1F5 scFv-Ig, followed by incubation with a peroxidase-conjugated GAH second Ab. Daming. S., Oliver. W. P., Theta. T., Martha. S., Hayden, J. A. (1999). Characterization of scFv-Ig Constructs Generated from the Anti-CD20 mAb 1F5 Using Linker Peptides of Varying Lengths1. J Immunol 1 June, 162 (11), 6589-6595. |
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