Probing effects of pressure release on virus capture during virus filtration using confocal microscopy

Shudipto K. Dishari, Adith Venkiteshwaran, Andrew L. Zydney

Research output: Contribution to journalArticlepeer-review

28 Scopus citations

Abstract

Virus filtration is used to ensure drug safety in the production of biotherapeutics. Several recent studies have shown a dramatic decrease in virus retention as a result of a process disruption, e.g., a transient pressure release. In this work, a novel two-label fluorescence technique was developed to probe virus capture within virus filtration membranes using confocal microscopy. Experiments were performed with Ultipor® DV20, Viresolve® Pro, and Viresolve® NFP membranes using bacteriophage ϕx174 as a model virus. The filters were challenged with two batches of fluorescently labeled phage: one labeled with red dye (Cy5) and one with green dye (SYBR Gold) to visualize captured phage from before and after the pressure release. The capture patterns seen in the confocal images were a strong function of the underlying membrane morphology and pore structure. The DV20 and Viresolve® NFP showed migration of previously captured phage further into the filter, consistent with the observed loss of virus retention after the pressure release. In contrast, there was no migration of captured virus in the Viresolve® Pro membranes, and these filters were also the only ones to show stable virus retention after a pressure release. The direct visualization of virus capture using the two-label fluorescence technique provides unique insights into the factors controlling the retention characteristics of virus filters with different pore structure.

Original languageEnglish (US)
Pages (from-to)2115-2122
Number of pages8
JournalBiotechnology and Bioengineering
Volume112
Issue number10
DOIs
StatePublished - Oct 1 2015

Keywords

  • Bioprocessing
  • Confocal microscopy
  • Pressure release
  • Virus clearance
  • Virus filtration

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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