SBIR-STTR Award

Life sciences research and other critical bioanalytical applications would strongly benefit from faster and higher resolution liquid chromatographic (HPLC) separations for both small molecules and larger molecules such as proteins, peptides, glycopeptides, and glycans. Many HPLC separations of biomedical interest, such as proteomic or glycomic profiling, require hours or days for resolution of a single sample. 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 various complex bioanalytical workflows, 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 our proprietary Fused-Core silica platform technology to non-spherical superficially porous particals, including ellipsoids, providing significantly faster and higher resolution separations. Our objectives are to create 2.5-5 ?m diameter ellipsoidal superficially porous Fused-Core silica particles, and to load these materials efficiently into HPLC column formats. The goals of the proposed work could 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. The separations technology described will directly lead to useful products for which there is a significant technical and market demand.

Public Health Relevance Statement:


Public Health Relevance:
High pressure liquid chromatography is the most widely used analytical method to separate mixtures of molecules, allowing measurement of quantities and identities of materials in a mixture. This method is broadly used in biomedical research, as well as in the creation, manufacture and control of therapeutic interventions. The current proposal is to use new knowledge in materials science and chemistry to enable faster and more efficient separations by liquid chromatography, saving time and money, as well as enabling new uses of the method to understand the structure and function of biological molecules.

Project Terms:
analytical method; Area; Biochemical; Biological; Biological Markers; Biological Process; Biological Products; Biological Sciences; Biomedical Research; Caliber; Chemistry; Chromatography; Complex; Deposition; Detection; Development; Dimensions; Disease; drug discovery; Evaluation; Evolution; Family; Genetic; Glycopeptides; Goals; High Pressure Liquid Chromatography; Hour; improved; Inferior; interest; Kinetics; Knowledge; Laboratories; Lead; Liquid Chromatography; Liquid substance; Literature; Marketing; Mass Spectrum Analysis; Measurement; Methods; Molecular Structure; novel; Nucleosome Core Particle; particle; Particle Size; Peptides; Performance; Permeability; Pharmacologic Substance; Phase; Polysaccharides; Post-Translational Protein Processing; Process; protein expression; Proteins; Proteomics; public health relevance; Research; Resolution; Rods (Retina); Sampling; Science; self assembly; Shapes; Silicon Dioxide; small molecule; Solid; Structure; System; Technology; Therapeutic Intervention; Thick; Time; Work

Extending liquid chromatographic column technology to even higher performance levels has given rise to ultra-high pressure liquid chromatography (UPLC), core-shell particle technology and instrumental developments such as lower volumes for injector and detector hydraulics. In spite of these advances, there is still room for improvement in speed, selectivity and resolution of the liquid chromatographic process. We propose that another level of improvement can be obtained with a change in particle shape by using ellipsoidal particles. These particles offer a reduced pressure drop, a higher mass fraction per unit volume of particles and the possibility to minimize wall effects that are characteristic o packed beds of spherical particles. Furthermore, the possibility of extending this non-spherical particle technology to smaller particle size is important because smaller spherical particles, while offering reduced zone broadening offer larger pressure drops. At some point, the advantage of superficially-porous particle architecture diminishes as particle size is reduced below ≈1.5 µm. If a route to smaller superficially porous non-spherical particles can be devised which minimizes the deleterious pressure drop of spheres, then performance can be increased before the pressure drop causes insurmountable difficulties. New chromatographic particles will be synthesized with a solid ellipsoidal or spherocylinder-like core and then a porous layer will be deposited around the outside for chromatographic retention. We have demonstrated previously in Phase I that there are advantages to this structure with regards to pressure drop and this can be rationalized by bed structures and performance that resemble a monolithic column without the problems of radial inhomogeneity and wall-effect-laden zone broadening that are present in monolithic column technology. We think of the proposed bed structure as that from a "pourable monolith." The current proposal uses synthesis technology and process-scale technology that were discovered and refined during Phase I efforts where it was shown that improved performance can be obtained for larger spherocylinder-like particles that are comparable with smaller spherical particles. In this comparison both the non-spherical and spherical particles used core-shell technology which AMT has pioneered. Phase II will expand on this effort, with the purpose of delivering further improved materials and methods to a broader range of applications in small molecule separations, such as metabolomics, to large molecules, such as proteins, glycoproteins and glycans. The aim here is to not only increase chromatographic resolution, but to make faster separations possible.

Public Health Relevance Statement:


Public Health Relevance:
High pressure liquid chromatography is the most widely used analytical method to separate mixtures of molecules, allowing measurement of quantities and identities of materials in a sample. This method is broadly used in biomedical research, as well as in the creation, manufacture and control of therapeutic interventions. The current proposal is to use new knowledge in materials science and chemistry to enable faster and more efficient separations by liquid chromatography, saving time and money, as well as enabling new uses of the method to understand the structure and function of biological molecules.

Project Terms:
analytical method; Architecture; base; Beds; Biological; Biological Process; Biological Sciences; Biomedical Research; Caliber; Characteristics; Chemistry; Deposition; detector; Development; Dimensions; Drops; Equipment; fitness; functional improvement; Glycoproteins; Goals; Height; High Pressure Liquid Chromatography; improved; Industry; interest; Knowledge; Laboratories; Lead; Liquid Chromatography; Liquid substance; macromolecule; Measurement; metabolomics; Methods; novel; Outcome; particle; Particle Size; Performance; Permeability; Pharmaceutical Preparations; Phase; Polysaccharides; pressure; Process; Proteins; public health relevance; Radial; Resolution; Route; Sampling; Science; Shapes; Silicon Dioxide; small molecule; Solid; Speed (motion); Structure; success; System; Technology; Therapeutic Intervention; Thick; Time; ultra high pressure; Varia