Physical Stabilization of Norwalk Virus‐Like Particles**The views, opinions and/or findings contained in this report are those of the author(s) and should not be construed as an official Department of the Army position, policy or decision unless so designated by other documentation.

Kissmann, Julian; Ausar, Salvador F.; Foubert, Thomas R.; Brock, Julie; Switzer, Mary H.; Detzi, Edward J.; Vedvick, Thomas S.; Middaugh, C. Russell

doi:10.1002/jps.21315

Cited by 48 publications

(12 citation statements)

References 30 publications

(39 reference statements)

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“…At 12 months in 15% trehalose, between 10% and 40% VLPs stored at −20°C showed altered morphology (data not shown), whereas no changes were observed at −70°C (Figure 2B) compared to freshly made VLPs (Figure 2A). This corroborates work of others who used trehalose to stabilise Norwalk [17] and rotavirus VLPs [18] and influenza vaccines [19]. VLPs stored in 5 and 15% sorbitol were unstable after 3 and 6 months, respectively, when stored at −20°C or −70°C (Figure 2D).…”

Section: Resultssupporting

confidence: 89%

“…Polyols (including glycerol, sucrose, sorbitol and trehalose) are found in vascular plants, fungi and algae [16]: these are natural, non-harmful substances and therefore suitable for use in vaccine formulations as stabilisers. Preliminary studies involving the use of trehalose, sucrose and glycerol as stabilisers for Norwalk [17] and rotavirus VLPs [18] and an influenza subunit vaccine [19] showed they enhance the stability of both VLPs and proteins.…”

Section: Resultsmentioning

confidence: 99%

“…The ability of 3 readily available osmolytes (trehalose, sorbitol and sucrose) to maintain the protein in a non-aggregated state without affecting its structure, potency and function at specific temperatures was investigated. Two different concentrations of osmolytes were selected (5% and 15%) to cover the range over which stability of other VLPs has been tested [17,18]. The number of freeze-thaw cycles that a suitably formulated sample could withstand without degrading was also determined.…”

Section: Resultsmentioning

confidence: 99%

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Stability studies of HIV-1 Pr55gagvirus-like particles made in insect cells after storage in various formulation media

Lynch

Meyers

Williamson

et al. 2012

Virol J

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BackgroundHIV-1 Pr55gag virus-like particles (VLPs) expressed by baculovirus in insect cells are considered to be a very promising HIV-1 vaccine candidate, as they have been shown to elicit broad cellular immune responses when tested in animals, particularly when used as a boost to DNA or BCG vaccines. However, it is important for the VLPs to retain their structure for them to be fully functional and effective. The medium in which the VLPs are formulated and the temperature at which they are stored are two important factors affecting their stability.FindingsWe describe the screening of 3 different readily available formulation media (sorbitol, sucrose and trehalose) for their ability to stabilise HIV-1 Pr55gag VLPs during prolonged storage. Transmission electron microscopy (TEM) was done on VLPs stored at two different concentrations of the media at three different temperatures (4°C, –20°C and −70°C) over different time periods, and the appearance of the VLPs was compared. VLPs stored in 15% trehalose at −70°C retained their original appearance the most effectively over a period of 12 months. VLPs stored in 5% trehalose, sorbitol or sucrose were not all intact even after 1 month storage at the temperatures tested. In addition, we showed that VLPs stored under these conditions were able to be frozen and re-thawed twice before showing changes in their appearance.ConclusionsAlthough the inclusion of other analytical tools are essential to validate these preliminary findings, storage in 15% trehalose at −70°C for 12 months is most effective in retaining VLP stability.

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Section: Resultssupporting

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Stability studies of HIV-1 Pr55gagvirus-like particles made in insect cells after storage in various formulation media

Lynch

Meyers

Williamson

et al. 2012

Virol J

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“…Norovirus VLPs have potential use as vaccines, so these results indicate why nonionic surfactants such as Tween 20 could also be successful candidates for nontoxic vaccine excipients. 27 Conversely, both charged surfactants studied were able to disassemble VLPs at high concentration and solubilize them in surfactant micelles. These strong ionic surfactants are therefore good candidates for virus cleaners and disinfectants.…”

Section: Resultsmentioning

confidence: 94%

Characterization and control of surfactant-mediated Norovirus interactions

Mertens

Velev

2015

Soft Matter

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Understanding of the colloidal interactions of Norovirus particles in aqueous medium could provide insights on the origins of the notorious stability and infectivity of these widespread viral agents. We characterized the effects of solution pH and surfactant type and concentration on the aggregation, dispersion, and disassembly of Norovirus virus-like particles (VLPs) using dynamic light scattering, electrophoretic light scattering, and transmission electron microscopy. Owing to net negative surface charge of the VLPs at neutral pH, low concentrations of cationic surfactant tend to aggregate the VLPs, whereas low concentrations of anionic surfactant tend to disperse the particles. Increasing the concentration of these surfactants beyond their critical micelle concentration leads to virus capsid disassembly and breakdown of aggregates. Non-ionic surfactants, however, had little effect on virus interactions and likely stabilized them additionally in suspension. The data were interpreted on the basis of simple models for surfactant binding and re-charging of the virus capsid. We used zeta potential data to characterize virus surface charge and interpret the mechanisms behind these demonstrated surfactant-virus interactions. The fundamental understanding and control of these interactions will aid in practical formulations for virus inactivation and removal from contaminated surfaces.

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“…[24][25][26][27][28][29] Among these approaches the baculovirus expression system is straightforward and highly efficient for large-scale productions of highly purified VLPs for NoV vaccine development. [30][31][32][33][34] Noroviruses (Novs) are important pathogens causing epidemic acute gastroenteritis affecting millions of people worldwide. Due to the inability to cultivate Novs, current Nov vaccine development relies on bioengineering technologies to produce virus-like particles (vLPs) and other subviral particles of Novs as subunit vaccines.…”

Section: Advancement Of Nov-like Particle (Vlp) Vaccinementioning

confidence: 99%