1159425 Heldt The objective of this research is use high-throughput methods to understand the principles behind selective precipitation of viruses with osmolytes to create a universal purification method for viruses that uses measured values of virus self-interaction based on theoretical and not empirically derived correlations. Our hypothesis is that osmolytes will preferentially precipitate viruses due to preferential hydration (the selective partitioning of water around a solute molecule) and result in the aggregation of viruses due to hydrophobic interactions. Both enveloped and non-enveloped viruses will be examined to understand the differences in these viruses classes. This is a transformative method to approach virus removal. Most virus removal is done empirically, whereas in this proposal, we will quantify selfinteraction parameters that will guide future methods for virus purification, removal and detection. This will be accomplished through the completion of the project goals. Goal 1: Use high-throughput screening to discover precipitants that selectively precipitate an enveloped and non-enveloped virus. High-throughput screening in 96-well plates will allow quick evaluation of multiple precipitants and precipitation condition. Goal 2: Quantify the thermodynamics of viruses in contact with precipitants. Close examination of flocculent size and solubility of viruses with co-solvents will allow an in depth evaluation of virus self-interaction and virus hydrophobicity. Goal 3: Expand the virus range to determine if the precipitants can be used as a platform approach to virus precipitation. Two additional viruses will be examined to determine if precipitation conditions are universally applicable to multiple viruses. The proposed activities will also strengthen the research, education, and outreach at Michigan Tech, the local Great Lakes region, and throughout the world. The need to develop a platform approach to virus purification will increase the speed to market of new vaccines and this purification can also be applied to viral gene therapy vectors. This proposal will use osmolytes to probe the self-interaction of enveloped and non-enveloped viruses. The implications of this knowledge will far extend from the precipitation of viruses to expand the toolbox of any virus separation or detection method to allow a more theoretically based approach to the design of the technique. The included education plan will introduce students from primary education up to graduate students to the STEM fields and the need to understand virus interactions to create improved virus removal, purification, and sensing technologies. Participation in Summer Youth Programs (SYP) and the Western U.P. Science Fair will attract underrepresented and economically disadvantaged groups and bring them to Michigan Tech to experience hands-on science and engineering programs and encourage them to pursue a career in a STEM fields.
Precipitation and Self-Interaction of Viruses by Preferential Hydration