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Related Experiment Video

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Purification of High Yield Extracellular Vesicle Preparations Away from Virus
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Asymmetric flow field flow fractionation methods for virus purification.

Katri Eskelin1, Mirka Lampi1, Florian Meier2

  • 1Department of Biosciences and Institute of Biotechnology, Viikinkaari 9, University of Helsinki, FIN-00014 Helsinki, Finland.

Journal of Chromatography. A
|October 5, 2016
PubMed
Summary

Asymmetric flow field flow fractionation (AF4) offers a rapid and gentle method for purifying viruses like bacteriophage PRD1. This optimized technique also enables real-time analysis of virus production in infected cells.

Keywords:
Bacteriophage PRD1Icosahedral virusMembrane virusMonolithic chromatographyReal-time analysis of progeny virus productionUltracentrifugation

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Area of Science:

  • Virology
  • Biochemistry
  • Biophysics

Background:

  • High-quality viral preparations are crucial for biochemical and biophysical characterization.
  • Virus production and purification are typically laborious and time-consuming processes.

Purpose of the Study:

  • To optimize Asymmetric Flow Field Flow Fractionation (AF4) conditions for efficient purification of the model virus, bacteriophage PRD1.
  • To assess the suitability of AF4 for preparative scale virus purification and real-time analysis of virus production.

Main Methods:

  • Optimization of Asymmetric Flow Field Flow Fractionation (AF4) parameters for bacteriophage PRD1 purification.
  • Evaluation of AF4 performance based on virus recovery and specific infectivity.

Main Results:

  • Optimized AF4 conditions demonstrated suitability for PRD1 purification from diverse starting materials.
  • AF4 achieved high virus recovery and specific infectivity, confirming its efficacy.
  • The method's speed and high sample capacity facilitated preparative scale purification.
  • AF4 enabled rapid, real-time monitoring of progeny virus production in infected cells.

Conclusions:

  • Asymmetric Flow Field Flow Fractionation (AF4) is a well-suited, rapid, and gentle technique for purifying bacteriophage PRD1.
  • Optimized AF4 protocols support both preparative scale purification and real-time analysis of viral production.