Lentivirus (LV) Production

Lentiviral vectors are one of the most commonly used viral vectors for therapeutic purposes due to their specific functional properties, such as their ability to infect non-dividing cells and packaging capacity. The RNA-based vector has been designed to deliver genes to mammalian cells safely and effectively, and there are several therapies using lentiviral vectors currently under development and in human clinical trials. Developing an efficient process for producing a lentiviral vector-based therapy is not easy. Purification of these 80-120 nm envelope vectors is challenging due to their low stability.   

There are multiple strategies for developing lentiviral vectors, but most follow the same basic process, with some variation as needed.   

Manufacturers are increasingly looking to produce viral vectors for gene therapy in suspension cell cultures, as the pressure increases for increased scalability. The cell seed train for suspension cultures generally occurs in smaller bioreactors, which may also double as small batch production vessels.  As production increases, larger volume bioreactors are needed. Viral vector production in suspension culture usually requires additional testing and optimization. Our experienced team of applications experts can work with you to determine the best solution for your upstream processes.

Ion exchange purification is the next step in this downstream process, to remove empty capsids, DNA and host cell proteins (HCP).  Adsorptive membrane technology offers an efficient and effective option for this step.

Manufacturing for clinical purposes requires the production of highly pure and biologically active vectors that meet regulatory requirements. A last sterilizing grade (0.2 µm) filtration step is usually needed before product is sent for final filling.