The purification of large biomolecules is an important component of biological research and for the production of recombinant proteins that are being utilized for numerous applications in medicine. The conventional chromatographic media used for this purpose have large (greater than 5 nanometer) pores to accommodate these large molecules. As a consequence these media are extremely fragile and compressive, making their use in column chromatography and bioprocessing very difficult. Furthermore, in order to minimize the compressive pressure drops produced by buffer flow, the particle size of the media need to be large, typically more than 100 micrometers in diameter. The large size and the extremely small diffusion coefficients of large molecules within the pores of the particles, greatly reduces the efficiency of the separation processes. The goal of this project is focused on the development of a new type of chromatographic medium which can be prepared by applying very thin polymeric coatings (less than 6 micrometers) onto non-woven fabrics and having ion exchange groups that are suitable for the adsorption of large biomolecules. The resulting chromatographic media will have the desired combination of high efficiency and robustness, making their use as chromatographic media more effective, more efficient, and at lower cost than existing media for similar applications.
The specific aims are (1) develop a family of non-wovens with polyurea coatings having ion exchange functionalities, high capacities, and high rates of mass transfer for recombinant proteins, monoclonal antibodies, and smaller proteins, (2) characterize the coated non-wovens in terms of their capacities, kinetics, and diffusivities for biomolecules using flow through devices, and (3) develop designs and build prototypes for chromatographic devices based on the properties of the coated non-wovens. ? ?
