Quantifying Lentiviral Physical Titer Using The QIAcuity dPCR System

By Miroslav Vraneš, Anne Schieren, Philipp Rink, Corinna Hochstein, Jonathan Thorn, Mario Quinto, Julius Albers, Domenica Martorana, Andreas Hecker, Ha minh Tri Pham, Anida Mesihovic Karamitsos, and Francesca Di Pasquale, QIAGEN

Developing safe and effective cell and gene therapies is critical to advancing treatments for a wide range of diseases. At the heart of many of these therapies lies the use of lentiviral vectors (LVs), a powerful and reliable tool for delivering therapeutic genes into target cells. However, to ensure the safety, consistency, and efficacy of these therapies, lentiviral preparations must undergo stringent quality control processes. This includes the accurate quantification of viral vector genome titers, determination of vector copy number (VCN), and detection of impurities.

Digital PCR (dPCR) has emerged as a superior method for these tasks, which offers unmatched precision, accuracy, and reproducibility compared to traditional quantitative PCR (qPCR). In response to the need for more efficient workflows, we have developed a direct reverse transcription digital PCR (RT-dPCR) approach for estimating LV titers. This one-step method, enabled by the CGT Lentivirus Lysis Kit, allows titering directly from crude lysates without RNA purification. This innovative method eliminates the need for RNA extraction or purification, yet delivers performance on par with conventional RT-dPCR techniques that rely on purified RNA. The workflow has been validated across various sample types and demonstrates robust performance, even after extended lysis incubation. Our streamlined dPCR workflow significantly enhances the characterization of lentiviral vectors during biopharmaceutical manufacturing.

It is especially suited for high-throughput environments and helps reduce turnaround times in QC and process development settings. Take the next step in advancing your gene therapy development—explore how our direct RT-dPCR solution can elevate your lentiviral vector analytics and accelerate your path to safe, effective treatments.