Lentiviral vectors (LVVs) are increasingly used in treatment of acquired and inherited genetic diseases. Production and purification of the LVVs remains challenging due to low titers from upstream processes and the low stability of the virus. A typical LVV purification strategy might include an AEX step positioned after clarification and before tangential flow filtration and final filtration. Thus, anion-exchange (AEX) chromatography has a key role in purification of LVVs. As the AEX step is positioned directly after clarification, relatively large volumes have to be processed. This makes membrane chromatography an ideal choice as it can be operated at and above 10 membrane volumes per minute. Despite widespread application of AEX membrane chromatography, there has been little published effort to understand the scalability of membrane devices when transitioning from filter plate experiments to small and then larger capsules. Here we implemented a Design of Experiment (DOE) approach to optimize the bind and elute conditions of LVVs in Mustang Q filter plate experiments and then transferred these conditions to 0.18 mL and 1 mL Mustang XT Acrodisc® Units. Gradient and step elution experiments were carried out to understand the role of salt and additives in desorption of LVV from the membrane surface. In addition, we compared the performance of Mustang Q devices to other AEX sorbents. Under optimized conditions, the observed capacity of Mustang Q capsules was 2.4 x 1011 to 1.5 x 1012 transfecting particles/mL of membrane. In summary, a scalable LVV purification protocol was developed for facile transition from Mustang Q filter plate to larger XT capsules.

Understanding scalability of Mustang® Q devices in lentiviral vector purification using DOE approach

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.