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 (1.7μmol/cm(2)) or medium (2.4μmol/cm(2)) density, it is possible to increase the recovery of purified virus by 60% compared to a highly charged membrane (3.3μmol/cm(2)) 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.
Keywords: Adenovirus adsorption; Hydrogel grafting; Ligand density; Membrane chromatography; Virus purification.
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