mOKT3-scFv[Puro] Lentivirus

The mOKT3-scFv[Puro] lentivirus is a ready-to-use, research-grade lentiviral preparation engineered to deliver a membrane-anchored single-chain variable fragment (scFv) derived from the murine anti-CD3 monoclonal antibody OKT3, and includes a puromycin resistance cassette for antibiotic selection. This vector is based on a third-generation lentiviral backbone, enabling stable genomic integration and persistent, consistent OKT3 scFv surface expression in a wide range of transfectable cell types, including many primary and difficult-to-transfect cells. The membrane-bound form restricts the scFv to the plasma membrane, promoting localized and sustained binding to CD3 on adjacent T cells while minimizing dilution or loss associated with secreted forms. The co-expressed puromycin marker allows for rapid enrichment of positive cells and long-term maintenance of engineered cell populations, simplifying the establishment of homogeneous cell lines and the construction of reproducible assay systems. By integrating antigen-binding capability and selection functionality into a single vector, this system reduces workflow complexity and variability compared to multi-vector approaches.

This construct is highly suitable for a variety of applications in T cell biology, synthetic immunology, and immuno-oncology research. Engineered cells expressing surface OKT3 scFv can serve as versatile artificial antigen-presenting cells or feeder cells to drive CD3-dependent T cell activation and expansion in vitro, enabling the study of proximal TCR/CD3 signaling events, downstream transcriptional programs, and functional readouts such as cytokine release and cytotoxicity. This platform supports the construction of controlled co-culture systems, mimicking immune synapse formation, and allowing for the dissection of variables in specific contexts, including the roles of co-stimulatory ligands, adhesion molecules, or inhibitory checkpoints when combined with other engineered features. In drug discovery, stable OKT3-scFv expressing cell lines can be used to evaluate small molecule or biologic modulators of T cell activation, analyze structure-function relationships, and assess candidate therapies affecting CD3-mediated signaling.