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

The scFv(CEA)-CD28-OX40-CD3zeta CAR-T lentivirus is a third-generation self-inactivating lentiviral vector designed to deliver a clinically inspired chimeric antigen receptor (CAR) comprising an anti-CEA single-chain variable fragment fused to CD28 and OX40 co-stimulatory domains and a CD3 zeta signaling motif. This vector is built on a modern lentiviral backbone, typically employing separate packaging and a SIN LTR structure, emphasizing genetic stability and safety, reducing the risk of promoter leakage and replication-competent lentivirus formation, while supporting durable and stable CAR expression in primary T cells. This vector design is valued for its ability to combine potent activation (CD28) with durability and memory support (OX40), ultimately driving robust T cell effector function through CD3ζ. This lentiviral platform broadly supports efficient transduction of human T cell subsets and stable genomic integration, which is particularly useful for downstream functional assays and longitudinal studies.

This lentiviral vector is suitable for generating CAR-T cells to assess their in vitro cytotoxicity against CEA-positive targets, analyze activation markers and cytokine release profiles, and explore T cell differentiation states, persistence, exhaustion, and memory formation under specific experimental conditions. Researchers can utilize it to study antigen density requirements and functional thresholds, compare the effects of co-stimulatory domains in parallel experimental designs, and map structure-function relationships of CAR signaling. It is commonly used in preclinical models related to colorectal cancer, gastric cancer, pancreatic cancer, and certain lung or breast cancers expressing CEA, enabling mechanistic studies of tumor cell killing mechanisms, resistance pathways, and the influence of the microenvironment on CAR-T function. The vector also supports the validation of combination therapy hypotheses—for example, studies involving co-administration with immune checkpoint inhibitors, targeted therapies, or cytokine support agents in a translational research setting—and can be used to develop and evaluate analytical workflows, including flow cytometry-based expression assessment, single-cell analysis, and transcriptomics.