Use of confocal scanning laser microscopy to study virus retention during virus filtration

Bakhshayeshi, Meisam; Jackson, Nigel B.; Kuriyel, Ralf; Mehta, Amit; Reis, Robert van; Zydney, Andrew L.

doi:10.1016/j.memsci.2011.05.069

Cited by 41 publications

(26 citation statements)

References 20 publications

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“…The depth of penetration was very similar at the two loadings, with the image at 44 L/m 2 showing a much greater fluorescent intensity. Additional details on the confocal microscopy are provided by Bakhshayeshi et al…”

Section: Results and Analysismentioning

confidence: 99%

“…0.5 mL of a 2.6 × 10 10 pfu/mL suspension of PP7 phage in 0.1 M bicarbonate buffer at pH 8.3 were mixed with one vial of the Alexa Fluor ® 488 dye and stirred overnight at 4°C. The labeled phage were then purified using a Bio‐Rad Bio‐Gel ® P‐30 fine size exclusion resin followed by diafiltration as described by Bakhshayeshi et al…”

Section: Methodsmentioning

confidence: 99%

“…A three‐dimensional fluorescent image of the filter was reconstructed by stacking the optical sections using the Olympus FluoView™ viewer software (Olympus American). Additional details are provided by Bakhshayeshi et al…”

Section: Methodsmentioning

confidence: 99%

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Internal virus polarization model for virus retention by the Ultipor^®VF Grade DV20 membrane

Jackson

Bakhshayeshi

Zydney

et al. 2014

Biotechnology Progress

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Several recent studies have reported a decline in virus retention during virus challenge filtration experiments, although the mechanism(s) governing this phenomenon for different filters remains uncertain. Experiments were performed to evaluate the retention of PP7 and PR772 bacteriophage through Ultipor VF Grade DV20 virus filters during constant pressure filtration. While the larger PR772 phage was fully retained under all conditions, a 2-log decline in retention of the small PP7 phage was observed at high throughputs, even under conditions where there was no decline in filtrate flux. In addition, prefouling the membrane with an immunoglobulin G solution had no effect on phage retention. An internal polarization model was developed to describe the decline in phage retention arising from the accumulation of phage in the upper (reservoir) layer within the filter which increases the challenge to the lower (rejection) layer. Independent support for this internal polarization phenomenon was provided by confocal microscopy of fluorescently labeled phage within the membrane. The model was in good agreement with phage retention data over a wide range of phage titers, confirming that virus retention is throughput dependent and supporting current recommendations for virus retention validation studies. These results provide important insights into the factors governing virus retention by membrane filters and their dependence on the underlying structure of the virus filter membrane.

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Section: Results and Analysismentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Internal virus polarization model for virus retention by the Ultipor^®VF Grade DV20 membrane

Jackson

Bakhshayeshi

Zydney

et al. 2014

Biotechnology Progress

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“…This asymmetry was also seen in the SEM images in Fig. 1 and in capture profiles for fluorescently-labeled bacteriophage imaged by confocal scanning laser microscopy (data not shown) [22]. Thus, when the membrane is oriented with the shiny-side down, the dextrans are able to enter the membrane pore structure more easily but are then retained within the interior of the filter, leading to a significant amount of internal concentration polarization in this orientation.…”

Section: Ultipor ® Dv20 Membranementioning

confidence: 64%

“…Baker) in deionized distilled water obtained from a NANOpure ® water and Ultipor ® DV20 (bottom) membrane. Reproduced from [22] with permission.…”

Section: Solution Preparationmentioning

confidence: 99%

Dextran sieving test for characterization of virus filtration membranes

Bakhshayeshi

Kanani

Mehta³

et al. 2011

Journal of Membrane Science

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Virus Filtration

Strauss,

Gudex,

Shirataki

2023

Kirk-Othmer Encyclopedia of Chemical Technology

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Biopharmaceutical products account for a large proportion of the recently developed medicinal drugs. Owing to their production in biological systems utilizing purified protein products from human blood plasma or recombinant production in cultured cells, biopharmaceuticals are at risk of virus contamination and, therefore, targeted virus removal steps are required in manufacturing processes for these products. Among several established methods for removing viruses, virus filtration has been widely adopted as the most reliable and robust method with effective clearance of a wide range of viruses. Here, we trace the history of the development of virus filters, describe the mechanism of virus removal, and explain the technical features of virus filters. Additionally, basic principles for using virus filters, including necessary user integrity tests and process validation are provided along with specific examples of current applications and new trends in their implementation.

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Use of confocal scanning laser microscopy to study virus retention during virus filtration

Cited by 41 publications

References 20 publications

Internal virus polarization model for virus retention by the Ultipor^®VF Grade DV20 membrane

Internal virus polarization model for virus retention by the Ultipor^®VF Grade DV20 membrane

Dextran sieving test for characterization of virus filtration membranes

Virus Filtration

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Use of confocal scanning laser microscopy to study virus retention during virus filtration

Cited by 41 publications

References 20 publications

Internal virus polarization model for virus retention by the Ultipor®VF Grade DV20 membrane

Internal virus polarization model for virus retention by the Ultipor®VF Grade DV20 membrane

Dextran sieving test for characterization of virus filtration membranes

Virus Filtration

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Product

Resources

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Internal virus polarization model for virus retention by the Ultipor^®VF Grade DV20 membrane

Internal virus polarization model for virus retention by the Ultipor^®VF Grade DV20 membrane