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Size exclusion removal of model mammalian viruses using a unique membrane system, Part I: Membrane qualification
A J DiLeo1, D A Vacante, E F Deane
1Millipore Corporation, Bedford, MA 01730.
Abstract:
We have previously described a new class of composite membrane that has the capability to efficiently remove particles, including viruses, from protein solution. The qualification of this membrane requires that it reproducibly and predictably remove model mammalian viruses. Using the Viresolve/70 membrane, the mammalian viruses polio, Simian virus-40, Sindbis, reovirus type 3 and Rauscher murine leukemia virus are shown to be reproducibly removed via a sieving mechanism. Mammalian virus retention increases constantly with virus diameter independent of virus class or type, increasing from 3.5 logs with polio virus to greater than 6.8 logs with murine leukemia virus. Consistent with a sieving mechanism, mammalian virus retention with the Viresolve/70 membrane is independent of virus concentration. These results are shown both in the presence and absence of protein in solution. The presence of protein in solution is shown to increase the virus retention coefficient of each virus above that measured in phosphate buffered saline. The model virus retention is shown to be well predicted by hard particle retention reported previously for this membrane. In addition, the hard particle retention is shown to predict the worse case performance expected of the membrane in the presence of protein.
Insights
This study demonstrates that the Viresolve/70 composite membrane effectively removes model mammalian viruses through a sieving mechanism. Virus retention is predictable based on diameter and unaffected by concentration, even in protein solutions.
Area of Science:
- Biotechnology
- Materials Science
- Virology
Background:
- Composite membranes offer efficient particle removal, including viruses, from protein solutions.
- Membrane qualification necessitates reproducible and predictable removal of model mammalian viruses.
Purpose of the Study:
- To evaluate the Viresolve/70 membrane's capability for reproducible mammalian virus removal.
- To elucidate the mechanism of virus retention by the composite membrane.
Main Methods:
- Utilized Viresolve/70 membrane for filtration studies.
- Employed model mammalian viruses: polio, Simian virus-40, Sindbis, reovirus type 3, and Rauscher murine leukemia virus.
- Assessed virus retention across varying concentrations and in the presence/absence of protein.
Main Results:
- Consistent, reproducible removal of all tested mammalian viruses via a sieving mechanism.
- Virus retention increased with diameter, ranging from 3.5 logs (polio virus) to over 6.8 logs (murine leukemia virus).
- Retention was independent of virus concentration and unaffected by protein presence, though protein slightly increased retention coefficients.
Conclusions:
- The Viresolve/70 membrane demonstrates predictable and efficient mammalian virus removal based on a sieving mechanism.
- Hard particle retention data accurately predicts membrane performance for virus removal, even in complex protein solutions.