Tie line framework to optimize non-enveloped virus recovery in aqueous two-phase systems

Joshi, Pratik U.; Turpeinen, Dylan G; Weiss, Matthew J.; Escalante-Corbin, Glendy; Schroeder, Michael; Heldt, Caryn L.

doi:10.1016/j.jchromb.2019.121744

Cited by 12 publications

(41 citation statements)

References 81 publications

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“…Appropriate amounts of stock solutions with 0.1 g crude virus were added to a microcentrifuge tube to total a 1 g system. The remaining procedure was similar to our study reported earlier (Joshi et al, 2019). The virus and VLP recoveries and partition coefficient ( K ) were calculated using Equations (1) and (2), respectively.

% Recovery = \frac{V_{P / C} \times T_{P / C}}{V_{i} \times T_{i}} \times 100

K = \frac{V_{P} \times T_{P}}{V_{C} \times T_{C}}

where V refers to the volume of either PEG‐rich phase ( P ), citrate‐rich phase ( C ), or initial stock ( i ) and T refers to the titer of the virus or VLP in either phases or initial stock.…”

Section: Methodssupporting

confidence: 69%

“…Appropriate amounts of stock solutions with 0.1 g crude virus were added to a microcentrifuge tube to total a 1 g system. The remaining procedure was similar to our study reported earlier (Joshi et al, 2019). The virus and VLP recoveries and partition coefficient (K) were calculated using Equations ( 1) and (2), respectively.…”

Section: Virus and Vlp Quantificationmentioning

confidence: 99%

“…Porcine parvovirus (PPV) strain NADL-2, gifted by Dr. Ruben Carbonell (North Carolina State University), was propagated in PK-13 cells as described previously (Heldt et al, 2006). PPV titration was performed by a colorimetric cell viability assay using MTT salt as described previously (Joshi et al, 2019).…”

Section: Cell Maintenance Virus Propagation and Virus Titrationmentioning

confidence: 99%

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Osmolyte enhanced aqueous two‐phase system for virus purification

Joshi

Turpeinen

Schroeder

et al. 2021

Biotech & Bioengineering

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Due to the high variation in viral surface properties, a platform method for virus purification is still lacking. A potential alternative to the high-cost conventional methods is aqueous two-phase systems (ATPSs). However, optimizing virus purification in ATPS requires a large experimental design space, and the optimized systems are generally found to operate at high ATPS component concentrations.The high concentrations capitalize on hydrophobic and electrostatic interactions to obtain high viral particle yields. This study investigated using osmolytes as driving force enhancers to reduce the high concentration of ATPS components while maintaining high yields. The partitioning behavior of porcine parvovirus (PPV), a nonenveloped mammalian virus, and human immunodeficiency virus-like particle (HIV-VLP), a yeast-expressed enveloped VLP, were studied in a polyethylene glycol (PEG) 12 kDa-citrate system. The partitioning of the virus modalities was enhanced by osmoprotectants glycine and betaine, while trimethylamine N-oxide was ineffective for PPV. The increased partitioning to the PEG-rich phase pertained only to viruses, resulting in high virus purification. Recoveries were 100% for infectious PPV and 92% for the HIV-VLP, with high removal of the contaminant proteins and more than 60% DNA removal when glycine was added. The osmolyte-induced ATPS demonstrated a versatile method for virus purification, irrespective of the expression system.

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% Recovery = \frac{V_{P / C} \times T_{P / C}}{V_{i} \times T_{i}} \times 100

K = \frac{V_{P} \times T_{P}}{V_{C} \times T_{C}}

Section: Methodssupporting

confidence: 69%

“…Appropriate amounts of stock solutions with 0.1 g crude virus were added to a microcentrifuge tube to total a 1 g system. The remaining procedure was similar to our study reported earlier (Joshi et al, 2019). The virus and VLP recoveries and partition coefficient (K) were calculated using Equations ( 1) and (2), respectively.…”

Section: Virus and Vlp Quantificationmentioning

confidence: 99%

Section: Cell Maintenance Virus Propagation and Virus Titrationmentioning

confidence: 99%

See 1 more Smart Citation

Osmolyte enhanced aqueous two‐phase system for virus purification

Joshi

Turpeinen

Schroeder

et al. 2021

Biotech & Bioengineering

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“…Viruses adsorb to surfaces through two main mechanisms, van der Waals (mainly mineral surfaces 10 ) and, more importantly, electrostatic interactions (charged surfaces in the presence of ions and or not neutral pH 11 , 12 , 13 ). These two forces dictate the adsorption of viruses to surfaces.…”

Section: Surface Stability Of Novel Coronavirus On Various Surfaces—amentioning

confidence: 99%

Surface Chemistry Can Unlock Drivers of Surface Stability of SARS-CoV-2 in a Variety of Environmental Conditions

Joonaki

Hassanpouryouzband

Heldt

et al. 2020

Chem

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119

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The surface stability and resulting transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), specifically in indoor environments, have been identified as a potential pandemic challenge requiring investigation. This novel virus can be found on various surfaces in contaminated sites such as clinical places; however, the behavior and molecular interactions of the virus with respect to the surfaces are poorly understood. Regarding this, the virus adsorption onto solid surfaces can play a critical role in transmission and survival in various environments. In this article, we first give an overview of existing knowledge concerning viral spread, molecular structure of SARS-CoV-2, and the virus surface stability is presented. Then, we highlight potential drivers of the SARS-CoV-2 surface adsorption and stability in various environmental conditions. This theoretical analysis shows that different surface and environmental conditions including temperature, humidity, and pH are crucial considerations in building fundamental understanding of the virus transmission and thereby improving safety practices.

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“…Virus surface chemistry, primarily the hydrophobicity and surface charge, determines its mobility and governs its colloidal behavior in virus adsorption processes [1,2]. Both the viral hydrophobicity and surface charge can dominate virus removal [3][4][5] and purification [6] performance in different solutions and influence viral persistence in the environment [7]. However, there is limited information on the viral surface chemistry [8][9][10][11].…”

mentioning

confidence: 99%

Single-Particle Chemical Force Microscopy to Characterize Virus Surface Chemistry

Heldt

2020

BioTechniques

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Two important viral surface characteristics are the hydrophobicity and surface charge, which determine the viral colloidal behavior and mobility. Chemical force microscopy allows the detection of viral surface chemistry in liquid samples with small amounts of virus sample. This single-particle method requires the functionalization of an atomic force microscope (AFM) probe and covalent bonding of viruses to a surface. A hydrophobic methyl-modified AFM probe was used to study the viral surface hydrophobicity, and an AFM probe terminated with either negatively charged carboxyl acid or positively charged quaternary amine was used to study the viral surface charge. With an understanding of viral surface properties, the way in which viruses interact with the environment can be better predicted.

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Tie line framework to optimize non-enveloped virus recovery in aqueous two-phase systems

Cited by 12 publications

References 81 publications

Osmolyte enhanced aqueous two‐phase system for virus purification

Osmolyte enhanced aqueous two‐phase system for virus purification

Surface Chemistry Can Unlock Drivers of Surface Stability of SARS-CoV-2 in a Variety of Environmental Conditions

Single-Particle Chemical Force Microscopy to Characterize Virus Surface Chemistry

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