Intellectual Merit: This EAGER proposal is a high risk high gain proposal. Recent outbreak of H1N1 flu and the worldwide spread of H5N1 avian influenza virus and its infection to human beings has raised a global concern about these viruses to cause a pandemic. According to the World Health Organization report of May 20, 2009, 41 countries have confirmed 10,243 cases of H1N1 flu with 80 fatalities. In the US, these numbers are 5,469 and 6, respectively. A treatment or a rapid vaccination process for H1N1 and H5N1 is urgently needed. The proposal addresses this issue in this transformative idea to develop a peptide-based membrane system so that flu viruses can be captured from either a cell culture broth or from human blood. The main goal is to develop a cell culture-based vaccine production method, which uses a peptide-based purification step. The peptide is developed using the available genomic information regarding hemagglutinin (HA) surface protein of flu viruses. We hypothesize that for a given virus, an affinity peptide with a high specificity could be selected by using a screening tool. Moreover, with the chemical attachment of the affinity peptide group to a membrane surface, this affinity matrix could be used to repeatedly bind and elute the specific type of virus. We further hypothesize that the virus surface protein plays a significant role on the virus binding process. Therefore, an affinity membrane designed based on a specific strain of influenza virus may be used to bind other viruses which have high homogeneity to this virus. This research will be helpful in better understanding the role of the virus surface protein on the affinity binding process. A term of 'homogeneity' is usually used to describe how similar two influenza virus strains are. By choosing several influenza viruses with different homogeneity, the effect of the surface protein on the virus binding to an affinity membrane could be characterized. This research will answer the question regarding how sensitive the effect of the homogeneity is on the binding of a virus to a given affinity membrane.
Since microporous affinity membrane combines both the advantages of the microfiltration and resin based affinity chromatography, the virus production and purification process could be significantly simplified. This is specifically suitable for the cell culture based influenza virus vaccine production process. By carefully choosing an appropriate membrane pore size, cells removal and virus purification could be achieved within one step. Cell cultures will be flown through one side of a microporous affinity membrane. During this period, cells and cell debris will be rejected by the membrane pores while viruses and proteins and/or DNA and other small biological components will migrate inside the membrane pores. Viruses will then be specifically bound by these functional peptide groups grafted on the membrane pore surface while protein, DNA and other biological components will pass through the membrane pores. After washing, viruses with high purity can be eluted from the bounded membranes. This simplification of the purification process may speed up the H1N1 and H5N1 vaccine development, commercialization and production process.
Broader Impact: Completion of the proposed studies will have significant impact and possible applications in H1N1 and H5H1 flu intervention. The proposed new peptide-based membrane will simplify the virus purification process and it may speed up the vaccine development, commercialization and production process. The prepared membrane may also be used in the virus diagnosis by quick collection and purification of virus samples. Further, this method may be extended to diagnose other viruses, such as the West Nile virus. Further, a possible clinical device may be developed which can specifically remove viruses from human blood. By circulating human blood through a membrane device containing affinity membranes for a specific flu virus, these viruses can be made to bind to the membrane surfaces thus reducing the overall virus population in the blood; this device may be then combined with some other intervention methods such as using TAMIFLU. This project will involve multidisciplinary researchers from Chemical Engineering and TTU Health Sciences Center. One of the educational goals of the project is to train one post-doc and one PhD student in the cutting edge research of vaccine and peptide-based membrane technology. An undergraduate student will be selected from well publicized outreach programs at TTU, e.g. Engineering Outreach Center, and McNair Scholars Program. The UG student will then be encouraged to go to graduate school and have a career in bio related field. The PI has significant experience in attracting female and minority candidates to his research group. Outreach to local schools will be done through funded programs such as Science it's a Girl's Thing, Pre-college Engineering, Teacher Training Workshop, and Engineering Outreach Mentor.
