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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
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.
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.
