title: Impact of grafting on the design of new membrane adsorbers for adenovirus purification.
abstract: 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.
authors.affiliation: IBET - Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal.
authors.forename: Piergiuseppe
authors.initials: P
authors.lastname: Nestola
authors.affiliation: Sartorius Stedim Biotech, August-Spindler-Strasse 11, 37079 Gottingen, Germany.
authors.forename: Louis
authors.initials: L
authors.lastname: Villain
authors.affiliation: IBET - Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal.
authors.forename: Cristina
authors.initials: C
authors.lastname: Peixoto
authors.affiliation: IBET - Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal.
authors.forename: Duarte L
authors.initials: DL
authors.lastname: Martins
authors.affiliation: IBET - Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal.
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
authors.affiliation: IBET - Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal; Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal; Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: Manuel J T
authors.initials: MJ
authors.lastname: Carrondo
authors.affiliation: Requimte/CQFB, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal; IBET - Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal. Electronic address: pmota@fct.unl.pt.
authors.forename: José P B
authors.initials: JP
authors.lastname: Mota
mesh_terms: D000256:Adenoviridae / Q000737:chemistry / Q000302:isolation & purification*; D000327:Adsorption; D002852:Chromatography, Ion Exchange; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D008024:Ligands; D008567:Membranes, Artificial
chemical_list: D008024:Ligands; D008567:Membranes, Artificial; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate
keywords: Adenovirus adsorption; Hydrogel grafting; Ligand density; Membrane chromatography; Virus purification