More Like this

1. Arginine‐enveloped virus inactivation and potential mechanisms

   https://doi.org/10.1002/btpr.2931  

   Meingast, Christa; Heldt, Caryn_L (November 2019, Biotechnology Progress)

   Abstract Arginine synergistically inactivates enveloped viruses at a pH or temperature that does little harm to proteins, making it a desired process for therapeutic protein manufacturing. However, the mechanisms and optimal conditions for inactivation are not fully understood, and therefore, arginine viral inactivation is not used industrially. Optimal solution conditions for arginine viral inactivation found in the literature are high arginine concentrations (0.7–1 M), a time of 60 min, and a synergistic factor of high temperature (≥40°C), low pH (≤pH 4), or Tris buffer (5 mM). However, at optimal conditions full inactivation does not occur over all enveloped viruses. Enveloped viruses that are resistant to arginine often have increased protein stability or membrane stabilizing matrix proteins. Since arginine can interact with both proteins and lipids, interaction with either entity may be key to understanding the inactivation mechanism. Here, we propose three hypotheses for the mechanisms of arginine induced inactivation. Hypothesis 1 describes arginine‐induced viral inactivation through inhibition of vital protein function. Hypothesis 2 describes how arginine destabilizes the viral membrane. Hypothesis 3 describes arginine forming pores in the virus membrane, accompanied by further viral damage from the synergistic factor. Once the mechanisms of arginine viral inactivation are understood, further enhancement by the addition of functional groups, charges, or additives may allow the inactivation of all enveloped viruses in mild conditions. 

   more » « less
2. Transfer Rate of Enveloped and Nonenveloped Viruses between Fingerpads and Surfaces

   https://doi.org/10.1128/AEM.01215-21  

   Anderson, Claire E.; Boehm, Alexandria B. (October 2021, Applied and Environmental Microbiology)

   Elkins, Christopher A. (Ed.)

   ABSTRACT Fomites can represent a reservoir for pathogens, which may be subsequently transferred from surfaces to skin. In this study, we aim to understand how different factors (including virus type, surface type, time since last hand wash, and direction of transfer) affect virus transfer rates, defined as the fraction of virus transferred, between fingerpads and fomites. To determine this, 360 transfer events were performed with 20 volunteers using Phi6 (a surrogate for enveloped viruses), MS2 (a surrogate for nonenveloped viruses), and three clean surfaces (stainless steel, painted wood, and plastic). Considering all transfer events (all surfaces and both transfer directions combined), the mean transfer rates of Phi6 and MS2 were 0.17 and 0.26, respectively. Transfer of MS2 was significantly higher than that of Phi6 ( P  < 0.05). Surface type was a significant factor that affected the transfer rate of Phi6: Phi6 is more easily transferred to and from stainless steel and plastic than to and from painted wood. Direction of transfer was a significant factor affecting MS2 transfer rates: MS2 is more easily transferred from surfaces to fingerpads than from fingerpads to surfaces. Data from these virus transfer events, and subsequent transfer rate distributions, provide information that can be used to refine quantitative microbial risk assessments. This study provides a large-scale data set of transfer events with a surrogate for enveloped viruses, which extends the reach of the study to the role of fomites in the transmission of human enveloped viruses like influenza and SARS-CoV-2. IMPORTANCE This study created a large-scale data set for the transfer of enveloped viruses between skin and surfaces. The data set produced by this study provides information on modeling the distribution of enveloped and nonenveloped virus transfer rates, which can aid in the implementation of risk assessment models in the future. Additionally, enveloped and nonenveloped viruses were applied to experimental surfaces in an equivalent matrix to avoid matrix effects, so results between different viral species can be directly compared without confounding effects of different matrices. Our results indicating how virus type, surface type, time since last hand wash, and direction of transfer affect virus transfer rates can be used in decision-making processes to lower the risk of viral infection from transmission through fomites. 

   more » « less
3. Thermostabilization of viruses via complex coacervation

   https://doi.org/10.1039/D0BM01433H  

   Mi, Xue; Blocher McTigue, Whitney C.; Joshi, Pratik U.; Bunker, Mallory K.; Heldt, Caryn L.; Perry, Sarah L. (December 2020, Biomaterials Science)

   null (Ed.)

   Widespread vaccine coverage for viral diseases could save the lives of millions of people each year. For viral vaccines to be effective, they must be transported and stored in a narrow temperature range of 2–8 °C. If temperatures are not maintained, the vaccine may lose its potency and would no longer be effective in fighting disease; this is called the cold storage problem. Finding a way to thermally stabilize a virus and end the need to transport and store vaccines at refrigeration temperatures will increase access to life-saving vaccines. We explore the use of polymer-rich complex coacervates to stabilize viruses. We have developed a method of encapsulating virus particles in liquid complex coacervates that relies on the electrostatic interaction of viruses with polypeptides. In particular, we tested the incorporation of two model viruses; a non-enveloped porcine parvovirus (PPV) and an enveloped bovine viral diarrhea virus (BVDV) into coacervates formed from poly(lysine) and poly(glutamate). We identified optimal conditions ( i.e. , the relative amount of the two polypeptides) for virus encapsulation, and trends in this composition matched differences in the isoelectric point of the two viruses. Furthermore, we were able to achieve a ∼10 3 –10 4 -fold concentration of virus into the coacervate phase, such that the level of virus remaining in the bulk solution approached our limit of detection. Lastly, we demonstrated a significant enhancement of the stability of non-enveloped PPV during an accelerated aging study at 60 °C over the course of a week. Our results suggest the potential for using coacervation to aid in the purification and formulation of both enveloped and non-enveloped viruses, and that coacervate-based formulations could help limit the need for cold storage throughout the transportation and storage of vaccines based on non-enveloped viruses. 

   more » « less
4. Inactivation of Coronaviruses and Phage Phi6 from Irradiation across UVC Wavelengths

   https://doi.org/10.1021/acs.estlett.1c00178  

   Ma, Ben; Linden, Yarrow S.; Gundy, Patricia M.; Gerba, Charles P.; Sobsey, Mark D.; Linden, Karl G. (March 2021, Environmental Science & Technology Letters)

   null (Ed.)

   Ultraviolet (UV) devices emitting UVC irradiation (200− 280 nm) have proven to be effective for virus disinfection, especially on surfaces and in air, due to their rapid effectiveness and limited to no material corrosion. Numerous studies of UV-induced inactivation focused on nonenveloped viruses. Little is known about UVC action on enveloped viruses across UVC wavelengths. In this study, we determined inactivation efficiencies of two coronaviruses (ssRNA) and an enveloped dsRNA bacteriophage surrogate in buffered aqueous solution (pH 7.4) using five commonly available UVC devices that uniquely emit light at different wavelengths spanning 222 nm emitting krypton chloride (KrCl*) excimers to 282 nm emitting UVC LEDs. Our results show that enveloped viruses can be effectively inactivated using UVC devices, among which the KrCl* excimer had the best disinfection performance (i.e., highest inactivation rate) for all three enveloped viruses. The coronaviruses exhibited similar sensitivities to UV irradiation across the UVC range, whereas the bacteriophage surrogate was much more resistant and exhibited significantly higher sensitivity to the Far UVC (<230 nm) irradiation. This study provides necessary information and guidance for using UVC devices for enveloped virus disinfection, which may help control virus transmission in public spaces during the ongoing COVID-19 pandemic and beyond. 

   more » « less
5. A replicon-based COVID-19 vaccine candidate delivered by tobacco mosaic virus-like particles

   https://doi.org/10.1016/j.vaccine.2025.127063  

   Karan, Sweta; Opdensteinen, Patrick; Ma, Yifeng; De_Oliveira, Jessica_Fernanda Affonso; Steinmetz, Nicole F (April 2025, Vaccine)

   The COVID-19 pandemic highlights the opportunity for mRNA vaccines and their nanotechnology carriers to make an impact as a countermeasure to infectious disease. As alternative to the synthetic lipid nanoparticles or mammalian viruses, we developed a tobacco mosaic virus (TMV)-based mRNA vaccine delivery platform. Specifically, purified coat protein from TMV was used to package a self-amplifying Nodamura replicon expressing the receptor binding domain (RBD) from the Omicron strain of SARS-CoV-2. The replicon construct contains the origin of assembly sequence from the tobacco mosaic virus (TMV) for encapsulation and mRNA stabilization. The nanoparticle vaccine was obtained through in vitro assembly using purified TMV coat proteins and in vitro transcribed mRNA cassettes. Cell assays confirmed delivery of self-amplifying mRNA vaccine, amplification of the transgene and expression of the target protein, RBD, in mammalian cells. Immunization of mice yielded RBDspecific IgG antibodies that demonstrated neutralization of SARS-CoV-2 using an in vitro neutralization assay. The TMV platform nanotechnology does not require ultralow freezers for storage or distribution; and the in vitro assembly method provide ‘plug-and-play’ to adapt the vaccine formulation rapidly as new strains or diseases emerge. Finally, opportunity exists to produce and self-assemble the vaccine candidate in plants through molecular farming techniques, which may allow production in the region-for the region and could make a contribution to less resourced areas of the world. 

   more » « less