scFv(EPCAM)-CD28-OX40-CD3zeta CAR-T Lentivirus

Lentiviral vectors have become the gold standard in CAR-T cell engineering due to their ability to stably integrate chimeric antigen receptor transgenes into the host cell genome, thereby achieving long-term stable protein expression across multiple generations of daughter cells. This particular vector employs a self-inactivating (SIN) long terminal repeat (LTR) design, significantly reducing the risk of insertional mutations and preventing the generation of replicative lentiviruses (RCLs) within the host cell. Compared to traditional retroviral vectors or non-viral transfection methods, these lentiviral particles exhibit superior transduction capabilities, effectively transducing both dividing and non-dividing cells, especially difficult-to-modify primary human T lymphocytes. High-titer production of these particles enables efficient gene delivery while minimizing cytotoxicity, thus maintaining the physiological state, stemness, and proliferative potential of T cells. Furthermore, the modular design of this vector is optimized to accommodate the large and complex gene sequences of third-generation CAR constructs—including scFv, hinge/transmembrane regions, and three distinct intracellular signaling domains—without affecting viral genome stability or overall infectious titer.

The primary application of this scFv(EPCAM)-CD28-OX40-CD3ζ CAR-T lentivirus lies in the development and preclinical evaluation of advanced cellular immunotherapies targeting EpCAM-positive solid tumors. EpCAM (epithelial cell adhesion molecule) is a well-characterized transmembrane glycoprotein highly expressed in various epithelial-derived cancers, including colorectal, gastric, breast, prostate, and lung cancers, while its expression is limited in normal tissues. By transducing T cells with this lentivirus, researchers can generate highly efficient effector cells that specifically recognize and eliminate malignant cells expressing EpCAM. These lentiviral particles are indispensable tools for in vitro cytotoxicity assays, cytokine release assays, and in vivo efficacy studies using humanized xenograft models. They enable detailed investigations of CAR-T cell migration, their ability to infiltrate dense tumor tissue, and potential off-target toxicity.