Comparison of the performance of anion exchange membrane materials for adenovirus purification using laterally-fed membrane chromatography

Kawka, Karina; Wilton, Amanda N.; Redmond, Eric J.; Medina, Maria Fe C.; Lichty, Brian D.; Ghosh, Raja; Latulippe, David R.

doi:10.1016/j.bej.2022.108417

Cited by 7 publications

(2 citation statements)

References 25 publications

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Section: Downstream Processing Efficiently Eliminates Wtad5mentioning

confidence: 99%

“…Centrifugation was followed by a lysis and clarification step. Mustang Q membrane adsorber was used as the first purification step since it is established that this ion exchange chromatography (IEX) membrane is able to efficiently retain wtAd5 (Kawka et al, 2022). To optimize rAAV2 recovery while maximizing wtAd5 binding to the membrane, the ionic strength of the loading buffer (containing NaCl concentrations ranging 200‐400 mM) was evaluated.…”

Section: Downstream Processing Efficiently Eliminates Wtad5mentioning

confidence: 99%

See 1 more Smart Citation

Towards a scalable bioprocess for rAAV production using a HeLa stable cell line

Escandell

Moura

Carvalho

et al. 2023

Biotech & Bioengineering

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The majority of recombinant adeno‐associated viruses (rAAV) approved for clinical use or in clinical trials areproduced by transient transfection using the HEK293 cell line. However, this platform has several manufacturing bottlenecks at commercial scales namely, low product quality (full to empty capsid ratio <20% in most rAAV serotypes), lower productivities obtained after scale‐up and the high cost of raw materials, in particular of Good Manufacturing Practice grade plasmid DNA required for transfection. The HeLa‐based stable cell line rAAV production system provides a robust and scalable alternative to transient transfection systems. Nevertheless, the time required to generate the producer cell lines combined with the complexity of rAAV production and purification processes still pose several barriers to the use of this platform as a suitable alternative to the HEK293 transient transfection. In this work we streamlined the cell line development and bioprocessing for the HeLaS3‐based production of rAAV. By exploring this optimized approach, producer cell lines were generated in 3‐4 months, and presented rAAV2 volumetric production (bulk) > 3 × 1011 vg/mL and full to empty capsids ratio (>70%) at 2 L bioreactor scale. Moreover, the established downstream process, based on ion exchange and affinity‐based chromatography, efficiently eliminated process related impurities, including the Adenovirus 5 helper virus required for production with a log reduction value of 9. Overall, we developed a time‐efficient and robust rAAV bioprocess using a stable producer cell line achieving purified rAAV2 yields > 1 × 1011 vg/mL. This optimized platform may address manufacturing challenges for rAAV based medicines.

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Section: Downstream Processing Efficiently Eliminates Wtad5mentioning

confidence: 99%

Section: Downstream Processing Efficiently Eliminates Wtad5mentioning

confidence: 99%