This Small Business Innovation Research (SBIR) Phase I project will develop a prototype gas-phase two-dimensional electrophoresis (GP2DE) instrument for analyzing protein structures in mixtures with less sample and higher throughput compared to traditional two-dimensional gel electrophoresis (2DGE). The instrument leverages the previously developed multi-dimensional ion mobility spectrometry (MDIMS) method for separating gas-phase protein ions based on molecular size and shape in both time and space with high-performance IMS. Understanding proteins' functions will be crucial to achieving the advances required for personalized medicine, which will be a strong driver of the proteomics field both scientifically and economically. Consequently, designing specialized therapies requires intricate knowledge of the complete protein structure. GP2DE will provide a rapid tool for detecting differences in protein structures using coupled ion mobility spectrometry analyses to give 2D gel electrophoresis-type separations in seconds to minutes. Increasing the throughput for analyses of protein structures addresses the inefficiencies in the labor intensive 2DGE process to build the structure knowledge base more rapidly. In particular, tools that can detect post-translational modifications of proteins, such as glycosylation or phosphorylation, will be valuable to proteomics. GP2DE can detect these structural changes easily because the structural differences will result in different ion mobilities.
The broader impact/commercial potential of this project, if successful, will provide a flexible, general purpose biomolecule analysis tool for researchers with limited budgets and/or facility space. This would be particularly useful for undergraduate laboratories where the shortened learning curve would allow for more productive research under typical time and resource constraints. The proposed gas-phase two-dimensional electrophoresis (G2DPE) evolves from advanced IMS technologies for structural size and shape analyses that are geared to be user-friendly. The market for proteomics research products has been projected to be several billion dollars in the next five years. GP2DE reduces the need to perform many more time consuming 2DGE analyses that have higher operating costs from labor and supplies, decreasing research costs and adding to laboratory productivity. Therefore, having another powerful tool for elucidating protein structural information will propel the rate of development of the next-generation of biotherapies better tailored to individual physiologies. The research conducted will be disseminated in peer-reviewed scientific journals and at scientific conferences to foster the adoption and advancement of multi-dimensional IMS methods for proteomics.
This Small Business Innovation Research (SBIR) Phase I project at Excellims Corp. proved the concept of using a different analysis method called ion mobility spectrometry (IMS) for a new user-friendly laboratory instrument to analyze peptide and protein structures in mixtures to create a new tool to help life science researchers. The IMS method is well known for its role as the scientific technique behind the commonly seen explosives detectors at airports. This particular SBIR project resulted in a prototype of the future instrument and in an initial data set showing the abilities of the method for analyzing some commonly studied biological molecules. The new method requires only a very small amount of sample, uses a much smaller volume of chemicals, and needs only air for doing analyses, making it more efficient and environmentally friendly. It offers higher throughput vs. some of the more well-established separation methods historically used in laboratories. The completed future system would offer a flexible, general means for biological sample analysis in a compact footprint that preserves precious working space. A short learning curve for the system would make it particularly useful for undergraduate and commercial research laboratories to increase their productivity. However, all types of research environments could benefit from having another tool at their disposal to better reach their goals. Personalized medicine has become a reachable goal as proteins’ functions become well understood. Consequently, designing specialized therapies requires intricate knowledge of the complete protein structure. Determining their role relies on structural information crucial to achieving the breakthroughs required for making it a reality. The new type of IMS developed through this research possesses strength in detecting molecules’ structural differences to give separations based on shape, size, and structure in seconds to minutes. Increasing the throughput of the many analyses needed to be performed as part of these advanced research and development efforts addresses inefficiencies in the currently used, more labor intensive processes. More importantly, discerning modifications of proteins will be valuable to finding their functions, a place where the new method excels because the structural differences cause them to separate differently. Reducing the need to perform many more time consuming, higher cost analyses creates savings in operating costs from labor and supplies, lowering research costs and speeding the return on investment. Thus, having another powerful tool for elucidating protein structural information encourages more rapid development of highly targeted cures.
