Overview of CAR-T Cell Generation Using Optimized Plasmid Design and Lentiviral Vector Production

Gene modified cell therapy such as CAR-T cells is one of the most promising advanced therapy medicinal products (ATMP) to fight against cancer. These therapies typically use lentiviral vectors to transfer a gene of interest and modify patient’s cells. All the raw material used during the lentiviral vector manufacturing must be carefully selected based on different parameters such as quality, performance, supplier, etc. 
In this project, we generated CAR-T cells based on different plasmid constructs expressing a fusion protein CAR to track transduction efficiency. We first built different plasmid constructs using the plasmid engineering service from Polyplus, then encapsulate this gene of interest using a lentiviral vector produced with FectoVIR^®-LV, transduce CD4+ CD8+ T cells from peripheral blood with this lentiviral vector, expand them for 12 days and finally test the efficiency of the transduced CAR-T cells using a killing assay.