Presentation Information
[P04-519]Sustainable Detergents for Higher AAV Yields and Effective Viral Inactivation
○Takao Ito1, Kakolie Banerjee2, Taylor Goldstone2, John Preston2, Phillip Szymanski2, Akshat Gupta2, Alice Antonello3, Corinne Miller2 (1. Merck Ltd. (An Affiliate of Merck KGaA, Darmstadt, Germany) (Japan), 2. MilliporeSigma (An Affiliate of Merck KGaA, Darmstadt, Germany) (USA), 3. Merck KGaA. (Germany))
Keywords:
Adeno-associated virus,Detergent,Lysis,Viral inactivation,Viral vector
Adeno–associated viruses (AAV) are an increasingly important gene therapy vector, commonly produced in HEK293 cells. Their recovery requires cell lysis using detergents to release intracellular particles and prevent aggregation by disrupting hydrophobic interactions with host–derived impurities. As biopharmaceutical manufacturing places greater emphasis on sustainability, interest has grown in eco–friendly detergent alternatives that maintain process performance while reducing environmental impact. This has driven the development of new detergents designed to support efficient bioprocessing together with improved environmental profiles. Higher AAV yield enables gene therapy manufacturers to increase productivity and reduce overall manufacturing costs. In AAV manufacturing, detergents can also serve dual purposes by inactivating enveloped viruses during cell lysis. In this study, we evaluate the new detergents and benchmark their viral inactivation performance against commonly used detergents.
AAV5–expressing HEK293 cell suspensions were used to evaluate detergent–assisted lysis and viral inactivation. Two newly developed detergents, Deviron®13–S9 (non–ionic) and Deviron® C16 (zwitterionic), were assessed for lysis efficiency and viral inactivation performance. Tween™ 20, a widely used non–ionic detergent, served as the benchmark control. To examine detergent performance, each detergent was tested at multiple concentrations. Following lysis, samples were incubated, clarified by centrifugation, and AAV capsid titers in the supernatant were quantified by ELISA. To assess the contribution of ionic strength to reducing aggregation and improving recovery, sodium chloride (NaCl) was included in parallel lysis conditions. Salt addition was selected due to its known ability to minimize AAV aggregation with host–derived impurities and thereby enhance capsid solubility. For viral inactivation studies, model enveloped viruses (xMuLV and PRV) were spiked into HEK293 cell suspensions prior to detergent treatment. Viral titers were quantified by TCID50 assays following incubation, and log reduction values (LRV) were calculated relative to no–detergent controls.
Both Deviron detergents produced higher AAV capsid recovery than Tween™ 20, reflecting improved lytic performance and reduced aggregation. Salt addition further enhanced capsid recovery, consistent with its known effect on minimizing AAV complexation with impurities. In viral inactivation assays, both Deviron detergents achieved robust inactivation, with LRV >5 for PRV and >6 for xMuLV, comparable to or exceeding performance of Tween™ 20.
This work highlights the efficacy of these detergents in enhancing AAV production while ensuring viral safety during manufacturing. Their dual functionality–improving capsid yield and providing effective viral inactivation–supports the broader adoption of sustainable, eco–friendly alternatives in gene therapy manufacturing processes.
AAV5–expressing HEK293 cell suspensions were used to evaluate detergent–assisted lysis and viral inactivation. Two newly developed detergents, Deviron®13–S9 (non–ionic) and Deviron® C16 (zwitterionic), were assessed for lysis efficiency and viral inactivation performance. Tween™ 20, a widely used non–ionic detergent, served as the benchmark control. To examine detergent performance, each detergent was tested at multiple concentrations. Following lysis, samples were incubated, clarified by centrifugation, and AAV capsid titers in the supernatant were quantified by ELISA. To assess the contribution of ionic strength to reducing aggregation and improving recovery, sodium chloride (NaCl) was included in parallel lysis conditions. Salt addition was selected due to its known ability to minimize AAV aggregation with host–derived impurities and thereby enhance capsid solubility. For viral inactivation studies, model enveloped viruses (xMuLV and PRV) were spiked into HEK293 cell suspensions prior to detergent treatment. Viral titers were quantified by TCID50 assays following incubation, and log reduction values (LRV) were calculated relative to no–detergent controls.
Both Deviron detergents produced higher AAV capsid recovery than Tween™ 20, reflecting improved lytic performance and reduced aggregation. Salt addition further enhanced capsid recovery, consistent with its known effect on minimizing AAV complexation with impurities. In viral inactivation assays, both Deviron detergents achieved robust inactivation, with LRV >5 for PRV and >6 for xMuLV, comparable to or exceeding performance of Tween™ 20.
This work highlights the efficacy of these detergents in enhancing AAV production while ensuring viral safety during manufacturing. Their dual functionality–improving capsid yield and providing effective viral inactivation–supports the broader adoption of sustainable, eco–friendly alternatives in gene therapy manufacturing processes.
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