Continuous purification of an enveloped and non-enveloped viral particle using an aqueous two-phase system

Turpeinen, Dylan G; Joshi, Pratik U.; Kriz, Seth A.; Kaur, Supreet; Nold, Natalie M.; O’Hagan, David; Nikam, Savita; Masoud, Hassan; Heldt, Caryn L.

doi:10.1016/j.seppur.2021.118753

Cited by 15 publications

(25 citation statements)

References 47 publications

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“…A thorough discussion regarding the applications of ATPSs in extracting VLPs is summarized in Table 2. Several studies had employed polymer/saltbased ATPSs to separate and purify VLPs, yielding remarkable recovery rates and separation efficiency of product [32,[59][60][61][62][63][64]. For instance, Benavides et al [59] utilized PEG/salt ATPSs to primary recover nonenveloped double-layered rotavirus-like particles (dlRLP) expressed with insect cell baculovirus vector.…”

Section: Application Of Aqueous Two-phase Systems For Virus-like Part...mentioning

confidence: 99%

“…The optimized system had successfully yielded 99% of HIV‐VLPs in the PEG‐rich top phase with high removal of host cell proteins and DNA. In addition, the continuous and batch ATPS showed similar VLPs recovery yield, which infer that ATPS possesses attracting features of scalability and continuous manner of operation besides preventing loss in product purity or yield [64]. Another work on osmolyte‐enhanced PEG 12000/citrate ATPS for purification of HIV‐VLPs showed prominent result of high VLPs recovery yield in the PEG‐rich top phase as well as high removal of host cell proteins and DNA.…”

Section: Application Of Aqueous Two‐phase Systems For Virus‐like Part...mentioning

confidence: 99%

“…Therefore, the continuous manufacturing is suggested as an alternative processing pipeline to separate recombinant VLPs. So far, there is only one study that reported the continuous manufacturing with ATPS for HIV‐VLPs purification, prominent separation efficiency, and recovery yield of targeted product were achieved [64]. The continuous ATPS for VLPs processing is a trend and further investigation is suggested as a next‐generation process.…”

Section: Challenges and Prospectivesmentioning

confidence: 99%

“…A thorough discussion regarding the applications of ATPSs in extracting VLPs is summarized in Table 2 . Several studies had employed polymer/salt‐based ATPSs to separate and purify VLPs, yielding remarkable recovery rates and separation efficiency of product [32, 59–64]. For instance, Benavides et al.…”

Section: Application Of Aqueous Two‐phase Systems For Virus‐like Part...mentioning

confidence: 99%

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Downstream processing of virus‐like particles with aqueous two‐phase systems: Applications and challenges

Leong

Show

et al. 2022

J of Separation Science

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Section: Application Of Aqueous Two-phase Systems For Virus-like Part...mentioning

confidence: 99%

Section: Application Of Aqueous Two‐phase Systems For Virus‐like Part...mentioning

confidence: 99%

Section: Challenges and Prospectivesmentioning

confidence: 99%

Section: Application Of Aqueous Two‐phase Systems For Virus‐like Part...mentioning

confidence: 99%

See 2 more Smart Citations

Downstream processing of virus‐like particles with aqueous two‐phase systems: Applications and challenges

Leong

Show

et al. 2022

J of Separation Science

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show abstract

“…One solution to tackle the downstream hurdles is to purify viral particles with aqueous two‐phase systems (ATPSs). Viral purification using ATPS has demonstrated high yields and continuous operation capability in bioprocessing (Du et al, 2019; Ladd Effio et al, 2015; Turpeinen et al, 2021). However, this technique still requires extensive study to understand viral behavior in these types of biphasic systems.…”

Section: Introductionmentioning

confidence: 99%

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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Vaccine process technology—A decade of progress

Buckland,

Sanyal,

Ranheim

et al. 2024

Biotech & Bioengineering

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In the past decade, new approaches to the discovery and development of vaccines have transformed the field. Advances during the COVID‐19 pandemic allowed the production of billions of vaccine doses per year using novel platforms such as messenger RNA and viral vectors. Improvements in the analytical toolbox, equipment, and bioprocess technology have made it possible to achieve both unprecedented speed in vaccine development and scale of vaccine manufacturing. Macromolecular structure‐function characterization technologies, combined with improved modeling and data analysis, enable quantitative evaluation of vaccine formulations at single‐particle resolution and guided design of vaccine drug substances and drug products. These advances play a major role in precise assessment of critical quality attributes of vaccines delivered by newer platforms. Innovations in label‐free and immunoassay technologies aid in the characterization of antigenic sites and the development of robust in vitro potency assays. These methods, along with molecular techniques such as next‐generation sequencing, will accelerate characterization and release of vaccines delivered by all platforms. Process analytical technologies for real‐time monitoring and optimization of process steps enable the implementation of quality‐by‐design principles and faster release of vaccine products. In the next decade, the field of vaccine discovery and development will continue to advance, bringing together new technologies, methods, and platforms to improve human health.

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Continuous purification of an enveloped and non-enveloped viral particle using an aqueous two-phase system

Cited by 15 publications

References 47 publications

Downstream processing of virus‐like particles with aqueous two‐phase systems: Applications and challenges

Downstream processing of virus‐like particles with aqueous two‐phase systems: Applications and challenges

Osmolyte enhanced aqueous two‐phase system for virus purification

Vaccine process technology—A decade of progress

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