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Osmolyte enhanced aqueous two-phase system for virus purification.

Pratik U Joshi1,2, Dylan G Turpeinen1,2, Michael Schroeder1

  • 1Department of Chemical Engineering, Michigan Technological University, Houghton, Michigan, USA.

Biotechnology and Bioengineering
|June 15, 2021
PubMed
Summary

This study introduces osmolytes to enhance virus purification using aqueous two-phase systems (ATPSs). This method reduces component concentrations while achieving high yields for viruses like porcine parvovirus and HIV-VLP.

Keywords:
bioprocessingdownstream processinginterfacialvaccine

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Area of Science:

  • Biotechnology
  • Virology
  • Biochemistry

Background:

  • Virus purification faces challenges due to diverse viral surface properties, necessitating cost-effective platform methods.
  • Conventional purification techniques are often expensive, driving the need for alternatives like aqueous two-phase systems (ATPSs).
  • Optimizing ATPS for virus purification typically requires extensive experimentation and high component concentrations.

Purpose of the Study:

  • To investigate the use of osmolytes as enhancers to reduce ATPS component concentrations for virus purification.
  • To evaluate the efficacy of osmolyte-induced ATPS for purifying different types of viruses, including porcine parvovirus (PPV) and human immunodeficiency virus-like particles (HIV-VLPs).

Main Methods:

  • Utilized a polyethylene glycol (PEG) 12 kDa-citrate aqueous two-phase system.
  • Investigated the partitioning behavior of PPV and HIV-VLP in the presence of osmolytes (glycine, betaine, trimethylamine N-oxide).
  • Assessed the purification efficiency by measuring viral recovery and removal of contaminant proteins and DNA.

Main Results:

  • Osmolytes glycine and betaine significantly enhanced virus partitioning to the PEG-rich phase.
  • High purification was achieved for both PPV (100% recovery) and HIV-VLP (92% recovery).
  • Glycine addition resulted in high removal of contaminant proteins and over 60% DNA removal.

Conclusions:

  • Osmolyte-induced ATPS offers a versatile and efficient platform for virus purification, applicable across different viral types and expression systems.
  • This approach allows for reduced ATPS component concentrations while maintaining high viral yields and purity.
  • The method shows promise as a cost-effective alternative to conventional virus purification techniques.