Lentiviral Downstream Process Optimisation and Design of Experiments on CIM QA 0.05 mL Monolithic 96-Well Plates

Lentiviral vectors are being increasingly used as tools for stable integration of large gene inserts into the genomes of both dividing and non-dividing cells. With 3rd and 4th generation of lentiviral vectors being incompetent of replication, they also offer a relatively safe tool for more widespread use. Currently, lentiviral based therapies are primarily ex-vivo, such as CAR-T, however recent animal studies have successfully demonstrated their use in-vivo as well. To enable more widespread use, as well as development of in-vivo therapies in humans, better downstream processes for lentiviral purification will have to be developed.

In this work we present the results from initial buffer optimization and following DoEs, through which we successfully increased infectious lentiviral recovery. Interestingly, salt concentration was determined as the most important factor in loading sample for high infectious recovery of lentivirus from CIM QA column. We also present results from PATfix analytics, which we used to determine sample purity and composition.