Detection of Lentiviral Constructs for Release Testing of CAR-T Cells Using Digital Droplet PCR

The document describes the development and validation of a digital droplet PCR (ddPCR) method for detecting lentiviral constructs in CAR-T cell products. The aim was to ensure the absence of replication competent lentivirus (RCL) in these products, as required by FDA guidelines. The ddPCR method was shown to have several advantages over traditional qPCR, including improved limit of detection, elimination of the need for a standard curve, and reduced hands-on technical time. The study demonstrated the reproducibility, linearity, specificity, and sensitivity of the ddPCR method, making it a reliable test for the release of CAR-T products. The results confirmed that ddPCR could rapidly detect RCL, ensuring the safety of patient products. Future work will focus on adapting this technology to detect other targets in cell product manufacturing.