Life sciences research and other critical bioanalytical applications would strongly benefit fromfaster and higher resolution high performance liquid chromatographic (HPLC) separations of largermolecules including proteins, peptides, glycopeptides, and glycans. Proteins therapeutics are complexmolecules, susceptible to intended or unintended alterations of composition and conformation. Manytherapeutic proteins are inherently a mixture of closely related variants. Current practice is to employHPLC to resolve and quantify the components such protein mixtures, to assure identity and safety ofthe 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 complexityof samples, there are many examples in the current research literature that combine multiple HPLCanalyses to gain full details on protein composition, and to measure the levels of protein variants, bothduring 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 afundamental part of the analytical systems, and the time to achieve high resolution separations ofprotein samples is a great bottleneck. This proposal describes an approach to improve the separationefficiency of HPLC for protein analysis by extending the use of novel superficially porous particle(SPP) technologies with optimized characteristics, providing significantly faster and higher resolutionseparations of proteins in the native state. Our objectives are to create SPP silica particles,specifically design for use in conditions that maintain intact native structures, optimize the materialsproperties, and to load these materials efficiently into HPLC column formats. The proposed work willextend recent breakthroughs in material science that yield authentic improvements in currentmaterials, seeking to employ the technology in additional native modes of HPLC operation, includinghydrophobic interaction and ion exchange. The goals of the proposed work would yield very highperformance chromatographic products, greater than those currently available, to be applied broadlyin analysis of proteins, particularly for protein therapeutics. The application of the technology is notlimited to biopharmaceutical preparations, and, in fact, would benefit any current application that usesHPLC methods for larger biological molecule analyses. The separations technology described willdirectly 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: Architecture ; Engineering / Architecture ; Biological Products ; Biologic Products ; Biological Agent ; biopharmaceutical ; biotherapeutic agent ; Biological Sciences ; Biologic Sciences ; Bioscience ; Life Sciences ; Biomedical Research ; Chemistry ; Chromatography ; High Pressure Liquid Chromatography ; HPLC ; High Performance Liquid Chromatography ; High Speed Liquid Chromatography ; Liquid Chromatography ; Pharmaceutical Preparations ; Drugs ; Medication ; Pharmaceutic Preparations ; drug/agent ; Ethylene Oxide ; Oxirane ; Future ; Glycopeptides ; Glycoproteins ; Goals ; Heterogeneity ; Ion Exchange ; Lead ; Pb element ; heavy metal Pb ; heavy metal lead ; Libraries ; Literature ; Methods ; Molecular Conformation ; Molecular Configuration ; Molecular Stereochemistry ; conformation ; conformational state ; Particle Size ; Peptides ; Polysaccharides ; Glycans ; Proteins ; Quality Control ; Recombinant Proteins ; Research ; Safety ; Savings ; Science ; Silicon Dioxide ; Cristobalite ; Sand ; Silica ; Tridymite ; Technology ; Time ; Work ; Measures ; analytical method ; quality assurance ; improved ; Surface ; Phase ; Variant ; Variation ; Biological ; Evaluation ; Measurement ; Biological Process ; Biological Function ; Therapeutic ; fluid ; liquid ; Liquid substance ; Knowledge ; Hour ; Complex ; Reaction ; System ; interest ; experience ; particle ; Performance ; Isoforms ; Protein Isoforms ; success ; Speed ; Structure ; Hydrophobic Interactions ; novel ; intervention therapy ; Therapeutic Intervention ; Sampling ; Property ; drug development ; native protein drug ; pharmaceutical protein ; protein drug agent ; therapeutic protein ; Thickness ; Thick ; Diameter ; Caliber ; Address ; Measurable ; Protein Analysis ; Resolution ; Preparation ; Characteristics ; Molecular ; Modification ; Development ; developmental ; design ; designing ; next generation ; Outcome ; Therapeutic Monoclonal Antibodies ; MAb Therapeutics ; monoclonal antibody drugs ; therapeutic mAbs ; comparative ; product development ; operation ; materials science ; Antibody-drug conjugates ;
