Impact of grafting on the design of new membrane adsorbers for adenovirus purification

Piergiuseppe Nestola, Louis Villain, Cristina Peixoto, Duarte L Martins, P.M. Alves, Manuel José Teixeira Carrondo, José Paulo Barbosa Mota

Research output: Contribution to journalArticle

12 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)1-11
JournalJournal of Biotechnology
Volume181
DOIs
Publication statusPublished - 10 Jul 2014

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Adenoviridae
Purification
Membranes
Viruses
Hydrogel
Hydrogels
Recovery
Ligands
Benchmarking
Amines
Screening
Experiments
Throughput

Keywords

  • Adenovirus adsorption
  • Hydrogel grafting
  • Ligand density
  • Membrane chromatography
  • Virus purification

Cite this

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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({\circledR}) 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.",
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Impact of grafting on the design of new membrane adsorbers for adenovirus purification. / Nestola, Piergiuseppe; Villain, Louis; Peixoto, Cristina; Martins, Duarte L; Alves, P.M.; Carrondo, Manuel José Teixeira; Mota, José Paulo Barbosa.

In: Journal of Biotechnology, Vol. 181, 10.07.2014, p. 1-11.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Impact of grafting on the design of new membrane adsorbers for adenovirus purification

AU - Nestola, Piergiuseppe

AU - Villain, Louis

AU - Peixoto, Cristina

AU - Martins, Duarte L

AU - Alves, P.M.

AU - Carrondo, Manuel José Teixeira

AU - Mota, José Paulo Barbosa

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PY - 2014/7/10

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AB - 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.

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