Impact of grafting on the design of new membrane adsorbers for adenovirus purification
•Hydrogel grafted membranes are more suitable for virus purification than direct grafted ones.•Virus recovery is better for hydrogel grafted ligand densities of 1.6–2.7 μmol/cm2 and increased NaCl in the feed.•Direct grafting requires ligand densities >4.5 μmol/cm2 and NaCl concentrations <25 mM for improved virus recovery.•DNA's 10%-DBC increases 5-fold when the ligand density is increased from 1.7 to 3.3 μmol/cm2; this is not seen for virus.•A novel 96-well plate system coupled with a small-scale device minimizes the amount of bulk material used for this study.
The impacts of quaternary amine ligand density and matrix structure, namely hydrogel grafted and directly grafted, on state-of-the-art chromatographic membranes operated in bind-and-elute mode were evaluated for the purification of adenovirus serotype 5. The experiments were performed on a 96-well plate membrane holder, which is a convenient high-throughput screening tool for obtaining the best operating conditions for a process yield optimization. The results show that the hydrogel-grafted membranes are more suitable for virus purification than the directly grafted ones. By reducing the number of grafted ligands to low (1.7 μmol/cm2) or medium (2.4 μmol/cm2) density, it is possible to increase the recovery of purified virus by 60% compared to a highly charged membrane (3.3 μmol/cm2) that yielded a recovery rate lower than 30%. In the reported experiments, Sartobind® Q, chosen as benchmark comparison, provides a better compromise between high recovery and large dynamic binding capacity. Overall, this work contributes to the understanding and development of new membrane adsorbers specifically designed for virus purification.
Journal: Journal of Biotechnology - Volume 181, 10 July 2014, Pages 1–11