Addressing common challenges in lentiviral vector clarification using closed, automated counterflow centrifugation systems

The field of cell and gene therapy is rapidly growing. In particular, the use of lentiviruses in CAR-T applications is becoming of higher importance. Lentiviruses are enveloped viruses that are rather large (100-150nm) and are therefore very fragile due to the membrane surrounding the capsid produced when lentiviruses are budding from cells. Large enveloped viruses pose serious challenges during the downstream operation, leading to poor recovery and low infectivity and thus high manufacturing costs. One key challenge in the downstream process of lentivirus production is the clarification step where you need to separate this large virus from the cells that produced it.

 

In this webinar, we describe how the technique of counterflow centrifugation (CFC) provides a gentle but powerful technology for lentiviral vector clarification, allowing for effective separation of producing cells and lentiviral vectors by size. Additionally, we describe the benefits of using automated, closed counterflow centrifugation systems with gamma-irradiated consumables, as they provide the highest containment and product safety protection, while allowing for a better bioburden control strategy. We showcase two closed, automated cell processing systems, the Gibco™ CTS™ Rotea Counterflow Centrifugation System from Thermo Fisher Scientific and the Ksep400 from Sartorius, to demonstrate at different scales how they efficiently clarify lentiviral vector containing culture broth through the removal of cells, resulting in clarified output with strongly reduced turbidity, while maintaining high lentiviral titer upon recovery. These closed CFC systems bring a novel solution to the clarification of lentivirus, improving both containment and recovery yields.

 

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