scFv(CD33)-OX40-CD3zeta CAR-T Lentivirus

The architecture of CARs has evolved over the past few years with modifications to the intracellular domain. First-generation CARs contained only a signaling domain derived from the CD3 zeta chain. While these CARs could activate T cells, their in vivo anti-tumor activity was suboptimal due to low cytotoxicity and proliferation of T cells expressing these CARs. Second-generation CARs, in addition to the CD3 zeta signaling domain, also possess an intracellular costimulatory domain. As a result, T cells expressing these second-generation CARs have significantly improved in vivo proliferation, expansion, and activation persistence.

Creative Biogene''s CAR-expressing lentiviral vectors can be used to produce lentivirus and efficiently deliver the CAR expression cassette into T cells. The CAR-expressing lentiviral vector is first cloned in Escherichia coli. The CAR expression cassette contains the scFv domain, hinge region, transmembrane region, intracellular CD3 zeta signaling, and costimulatory domains, located between the lentiviral vector''s two long terminal repeats (LTRs). After construction, the lentiviral vector is transfected into packaging cells along with a helper plasmid. In the packaging cells, the DNA fragment between the two LTRs is transcribed into RNA, which is then packaged into viral particles by viral proteins expressed by the helper plasmid. The packaged live virus is released into the supernatant, which can be directly collected or further concentrated to transfect target cells.