The proposed product is an analytical scale, UHPLC column for separating protein drugs in native form, with the Phase I proposal focused on anion exchange chromatography as the separation mechanism. The bonded phase is made of polymer chains, in this case bearing weak anion exchange groups. The study will be expanded to cation exchange and hydrophobic interaction chromatography in future Phase II work. These non-denaturing separations are essential in the development of protein drugs because they allow the separated components to be tested for efficacy, immunogenicity and mechanism in phamacological research. Non- denaturing chromatography columns for protein drugs use polymeric bonded phases because these impart the molecular-scale flexibility needed for fast adsorption and desorption. The overall market size for non-denaturing chromatography columns is on the order of $1B, and the market leaders include Thermo-Dionex. Agilent, Waters and Sepax. The problems with current columns are two-fold: the separations are relatively low in resolution and the columns have short lifetimes. Preliminary results show both resolution column lifetime can be improved by an innovative manufacturing process. The latter considers that resolution and lifetime are limited because polymers are compressible, and the cycling between high pressure and ambient pressure through routine use of the column progressively disturbs the packing to degrade the separation over time. The proposed columns will be made by first tightly packing incompressible silica particles to give a pressure-invariant packing. Then the polymer bonded phase will be grown inside the column from initiators on the surfaces of the silica particles, forming a uniform bonded phase. The proposed work entails two specific aims. The first aim is to develop an automated procedure for in-column growth of the polymeric bonded phase. The second aim is to compare the column performance with those of leading commercial columns with respect to resolution of monoclonal antibodies having known charge heterogeneity and column lifetime. The outcome of the work will establish the feasibility of commercializing a line of non-denaturing chromatography columns for protein drugs. The concept will be extended in the future Phase II research to address scale-up issues in manufacturing and to develop other non-denaturing separations, including cation chromatography and hydrophobic interaction chromatography.
Public Health Relevance Statement: PROJECT NARRATIVE Ultrahigh performance liquid chromatography columns will facilitate development of protein drugs, particularly for cancer, with greater effectiveness and fewer side effects.
Project Terms: Address; Adsorption; Adverse effects; Anions; Back; Cations; Characteristics; Charge; Chromatography; Column Chromatography; Development; Drug usage; Effectiveness; efficacy testing; flexibility; Future; Goals; Growth; Heterogeneity; Hydrophobic Interactions; immunogenicity; improved; innovation; Length; Liquid Chromatography; Longitudinal Studies; Malignant Neoplasms; manufacturing process; Marketing; Measures; Modification; molecular scale; Monoclonal Antibodies; monolayer; Outcome; particle; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; polymerization; Polymers; pressure; Procedures; Process; Proteins; Reaction Time; Reproducibility; Research; Resolution; scale up; Silanes; Silicon Dioxide; Stainless Steel; statistics; Surface; Time; time use; Water; Work
