Chromatographic Purification of Virus Particles

Gagnon, Pete

doi:10.1002/9780470054581.eib583

Cited by 9 publications

(23 citation statements)

References 102 publications

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“…Lectins interact with accessible carbohydrate moieties of the surface glycoproteins according to their specificity. Heparin is primarily a weak and strong cation exchanger containing sulfo and carboxyl groups as well as numerous hydroxyl groups [113]. Desorption of the virus particles from heparin matrices is done by increasing the ionic strength.…”

Section: Surface Chemistry Of the Stationary Phase And The Composition mentioning

confidence: 99%

“…Desorption of the virus particles from heparin matrices is done by increasing the ionic strength. However, it often requires a higher ionic strength than for classical cation exchangers owing to the multivalent interactions of the branched linear structure of heparin and the formation of hydrogen bonding between the target virus and heparin [113]. An alternative to laborious ligand screens or the application of less specific pseudo-affinity ligands are engineered viral vectors containing affinity tags on the virus surface [109,114].…”

Section: Surface Chemistry Of the Stationary Phase And The Composition mentioning

confidence: 99%

“…Hydrophobic-interaction chromatography separates biomolecules dependent on the differential interaction of these compounds with hydrophobic ligands on the surface of the stationary phase. It is routinely used for bioseparations of proteins [119][120][121][122], and has also been applied for virus purification [100,113] since it is an orthogonal separation technique to purification methods based on ionic interactions. However, many factors such as type of salt and ionic strength of buffer, pH, temperature, ligand and ligand density influence the performance of HIC, making its development and optimization a difficult and time-consuming endeavor.…”

Section: Surface Chemistry Of the Stationary Phase And The Composition mentioning

confidence: 99%

“…However, many factors such as type of salt and ionic strength of buffer, pH, temperature, ligand and ligand density influence the performance of HIC, making its development and optimization a difficult and time-consuming endeavor. The two most commonly used HIC ligands are butyl and phenyl [113]. Interactions with these ligands are relatively strong and have been frequently cited to denature labile proteins, potentially reducing their efficacy.…”

Section: Surface Chemistry Of the Stationary Phase And The Composition mentioning

confidence: 99%

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Downstream processing of cell culture-derived virus particles

Wolf

Reichl

2011

Expert Review of Vaccines

121

102

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Manufacturing of cell culture-derived virus particles for vaccination and gene therapy is a rapidly growing field in the biopharmaceutical industry. The process involves a number of complex tasks and unit operations ranging from selection of host cells and virus strains for the cultivation in bioreactors to the purification and formulation of the final product. For the majority of cell culture-derived products, efforts focused on maximization of bioreactor yields, whereas design and optimization of downstream processes were often neglected. Owing to this biased focus, downstream procedures today often constitute a bottleneck in various manufacturing processes and account for the majority of the overall production costs. For efficient production methods, particularly in sight of constantly increasing economic pressure within human healthcare systems, highly productive downstream schemes have to be developed. Here, we discuss unit operations and downstream trains to purify virus particles for use as vaccines and vectors for gene therapy.

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Section: Surface Chemistry Of the Stationary Phase And The Composition mentioning

confidence: 99%

Section: Surface Chemistry Of the Stationary Phase And The Composition mentioning

confidence: 99%

Section: Surface Chemistry Of the Stationary Phase And The Composition mentioning

confidence: 99%

Section: Surface Chemistry Of the Stationary Phase And The Composition mentioning

confidence: 99%

See 2 more Smart Citations

Downstream processing of cell culture-derived virus particles

Wolf

Reichl

2011

Expert Review of Vaccines

121

102

View full text Add to dashboard Cite

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“…These virus characteristics can be utilized to fractionate them from other molecules [8]. The initial step in any purification process is to concentrate the molecules of interest.…”

Section: Introductionmentioning

confidence: 99%

Concentration and purification of enterovirus 71 using a weak anion-exchange monolithic column

Venkatachalam

Szyporta

Kiener

et al. 2014

Virol J

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BackgroundEnterovirus 71 (EV-71) is a neurotropic virus causing Hand, Foot and Mouth Disease (HFMD) in infants and children under the age of five. It is a major concern for public health issues across Asia-Pacific region. The most effective way to control the disease caused by EV-71 is by vaccination thus a novel vaccine is urgently needed. Inactivated EV-71 induces a strong, virus-neutralizing antibody response in animal models, protecting them against a lethal EV-71 challenge and it has been shown to elicit cross-neutralizing antibodies in human trials. Hence, the large-scale production of purified EV-71 is required for vaccine development, diagnosis and clinical trials.MethodsCIM® Monolith columns are single-piece columns made up of poly(glycidyl methacrylate co-ethylene dimethacrylate) as support matrix. They are designed as porous channels rather than beads with different chemistries for different requirements. As monolithic columns have a high binding capacity, flow rate and resolution, a CIM® DEAE-8f tube monolithic column was selected for purification in this study. The EV-71 infected Rhabdomyosarcoma (RD) cell supernatant was concentrated using 8% PEG 8000 in the presence of 400 mM sodium chloride. The concentrated virus was purified by weak anion exchange column using 50 mM HEPES + 1 M sodium chloride as elution buffer.ResultsHighly pure viral particles were obtained at a concentration of 350 mM sodium chloride as confirmed by SDS-PAGE and electron microscopy. Presence of viral proteins VP1, VP2 and VP3 was validated by western blotting. The overall process achieved a recovery of 55%.ConclusionsEV-71 viral particles of up to 95% purity can be recovered by a single step ion-exchange chromatography using CIM-DEAE monolithic columns and 1 M sodium chloride as elution buffer. Moreover, this method is scalable to purify several litres of virus-containing supernatant, using industrial monolithic columns with a capacity of up to 8 L such as CIM® cGMP tube monolithic columns.

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Downstream processing for influenza vaccines and candidates: An update

Carvalho

Peixoto

Carrondo

et al. 2021

Biotech & Bioengineering

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Seasonal and pandemic influenza outbreaks present severe health and economic burdens. To overcome limitations on influenza vaccines' availability and effectiveness, researchers chase universal vaccines providing broad, long-lasting protection against multiple influenza subtypes, and including pandemic ones.Novel influenza vaccine designs are under development, in clinical trials, or reaching the market, namely inactivated, or live-attenuated virus, virus-like particles, or recombinant antigens, searching for improved effectiveness; all these bring downstream processing (DSP) new challenges. Having to deal with new influenza strains, including pandemics, requires shorter development time, driving the development of faster bioprocesses. To cope with better upstream processes, new regulatory demands for quality and safety, and cost reduction requirements, new unit operations and integrated processes are increasing DSP efficiency for novel vaccine formats.This review covers recent advances in DSP strategies of different influenza vaccine formats. Focus is given to the improvements on relevant state-of-the-art unit operations, from harvest and clarification to purification steps, ending with sterile filtration and formulation. The development of more efficient unit operations to cope with biophysical properties of the new candidates is discussed: emphasis is given to the design of new stationary phases, 3D printing approaches, and continuous processing tools, such as continuous chromatography. The impact of the production platforms and vaccine designs on the downstream operations for the different influenza vaccine formats approved for this season are highlighted.

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Chromatographic Purification of Virus Particles

Cited by 9 publications

References 102 publications

Downstream processing of cell culture-derived virus particles

Downstream processing of cell culture-derived virus particles

Concentration and purification of enterovirus 71 using a weak anion-exchange monolithic column

Downstream processing for influenza vaccines and candidates: An update

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