SBIR-STTR Award

?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.

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.

NIH Spending Category:
Bioengineering; Biotechnology

Project Terms:
Address; analytical method; Area; Biological; Biological Process; Biological Products; Biological Sciences; Biomedical Research; Caliber; Characteristics; Chemistry; Complex; design; Development; Dimensions; Disease; Evaluation; Generations; Genetic; Glycopeptides; Glycoproteins; Goals; High Pressure Liquid Chromatography; Hour; improved; Individual; interest; Knowledge; Laboratories; Lead; Liquid Chromatography; Liquid substance; Literature; Marketing; Mass Spectrum Analysis; Measurement; meetings; Methods; Modification; new technology; novel; Outcome; particle; Particle Size; Peptides; Performance; Pharmacologic Substance; Polysaccharides; Post-Translational Protein Processing; product development; Property; protein expression; Proteins; public health relevance; Quality Control; Relative (related person); Research; Resolution; Safety; Sampling; Science; Silicon Dioxide; Structure; Surface; System; Technology; Therapeutic Agents; Therapeutic Intervention; Thick; Time; Work

Life sciences research and other critical bioanalytical applications would strongly benefit from faster and higher resolution high performance liquid chromatographic (HPLC) separations of larger molecules including proteins, peptides, glycopeptides, and glycans. Proteins therapeutics are complex molecules, susceptible to intended or unintended alterations of composition and conformation. Many therapeutic proteins are inherently a mixture of closely related variants. Current practice is to employ HPLC to resolve and quantify the components such protein mixtures, to assure identity and safety of the protein drug. Many HPLC separations of proteins are limited in resolution of protein mixtures, require an hour or more, and may not completely resolve all of the components. Due to the complexity of samples, there are many examples in the current research literature that combine multiple HPLC analyses to gain full details on protein composition, and to measure the levels of protein variants, both during development of the drug, as well as manufacture and quality assurance/quality control. Regardless of the intended use of the quantitative information, high efficiency LC separations are a fundamental part of the analytical systems, and the time to achieve high resolution separations of protein samples is a great bottleneck. This proposal describes an approach to improve the separation efficiency of HPLC for protein analysis by extending the use of novel superficially porous particle (SPP) technologies to larger pore size column packing materials of smaller size particles, with optimized characteristics, providing significantly faster and higher resolution separations of proteins. Our objectives are to create 2.0 ?m diameter and smaller SPP silica particles, optimize their materials properties, and to load these materials efficiently into HPLC column formats. The proposed work will extended recent breakthroughs in material science that yield authentic improvements in current materials, seeking to employ the technology in additional modes of HPLC operation, including hydrophobic interaction, ion exchange, and hydrophilic interaction. 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 proteins, particularly for protein therapeutics. The application of the technology is not limited to biopharmaceutical preparations, and, in fact, would benefit 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.

Public Health Relevance Statement:
Narrative 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:
Address; analytical method; Area; Biological; Biological Process; Biological Products; Biological Sciences; Biomedical Research; Caliber; Characteristics; Chemistry; Complex; Computer Simulation; design; Development; drug development; Environment; Evaluation; Exhibits; Future; Generations; Glycopeptides; Glycoproteins; Goals; Heterogeneity; High Pressure Liquid Chromatography; Hour; hydrophilicity; Hydrophobic Interactions; improved; interest; Ion Exchange; Knowledge; Laboratories; Lead; Liquid Chromatography; Liquid substance; Literature; materials science; Measurement; Measures; Mechanics; Methods; model design; Modification; Molecular; Molecular Conformation; novel; operation; Outcome; particle; Particle Size; Peptides; Performance; Permeability; Pharmaceutical Preparations; Phase; Polysaccharides; Preparation; product development; Production; Property; Protein Analysis; Protein Engineering; Proteins; quality assurance; Quality Control; Research; Resolution; Safety; Sampling; Savings; Science; Scientist; Silicon Dioxide; Structure; Surface; System; Technology; Therapeutic; Therapeutic Intervention; therapeutic protein; Thick; Time; Variant; Work