Improved Protein Separations with Superficially Porous Particles

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 DESCRIPTION (provided by applicant): Life sciences research and other critical bioanalytical applications would strongly benefit from faster and higher resolution liquid chromatographic (HPLC) separations of larger molecules including proteins, peptides, glycopeptides, and glycans. Many HPLC separations of biomedical interest, such as resolution of protein mixtures, require an hour or more, and may not completely resolve all of the components. Due to the complexity of biological samples, there are many examples in the current research literature that combine multiple dimensions of HPLC separations, often with online mass spectroscopy (MS). Although very time consuming, this approach has yielded useful information on protein expression and post-translational modifications, the biological processes underlying protein expression and modifications, and the effects thereon of development, disease state, and various environmental and genetic factors. In other cases, such as the development of protein therapeutic agents, many closely related protein molecules may be present, but precise knowledge of the individual identities and quantities of each could be required for assurance of safety and efficacy of the therapeutic agent. In either circumstance, high efficiency LC separations are a fundamental part of the analytical systems, and the time to achieve high resolution separations of complex biological samples is a great bottleneck. This proposal describes an approach to improve the separation efficiency of HPLC by extending the use of superficially porous particle (SPP) technologies to smaller size particles, with optimized characteristics, providing significantly faster and higher resolution separations. Our objectives are to create 2.0 µm diameter and smaller SPP silica particles, optimize their materials properties, and to load these materials efficiently into HPLC column formats. The goals of the proposed work would yield very high performance chromatographic products, greater than those currently available, to be applied broadly in analysis of complex biological samples, pharmaceutical and biopharmaceutical applications, in fact in any current application that uses HPLC methods for larger biological molecule analyses. The separations technology described will directly lead to useful products for which there is a significant technical and market demand.