SBIR-STTR Award

The product will be a packed capillary for nanoUHPLC of peptides that provides a dramatic increase in speed and efficiency. Current commercial columns for high speed and efficiency use sub-2 mm particles. The proposed technology uses sub-0.2 mm particles. The ability to use such small particles for UHPLC is enabled by the phenomenon of slip flow, which gives enhanced volume flow rates in reversed-phase liquid chromatography. Slip flow is a breakthrough that overcomes the fundamental limit to peak broadening caused by the parabolic velocity profile in the mobile phase. Using nonporous sub-0.2 mm particles also eliminates intraparticle diffusion while giving high phase ratios. Preliminary results show that sub micrometer nonporous silica particles reach the diffusion limit for efficiency in protein separations, and the diffusion limit is expected to be reached for the faster diffusing peptides. This gives plate numbers that are independent of length, allowing columns less than 2 cm in length to give 100,000 plates with high speed. The specific aims are to 1) optimize the particle diameter, 2) optimize the separation length, and 3) evaluate the nano LC-MS of a tryptic digest.

Public Health Relevance Statement:


Public Health Relevance:
A concept from Nano fluidics called slip flow is introduced to design chromatography columns having greatly improved speed for faster progress in disease diagnostics and therapy.

Project Terms:
base; Blood capillaries; Bovine Serum Albumin; Caliber; capillary; Chromatography; Column Chromatography; design; Diagnostic; Diffuse; Diffusion; Disease; Equilibrium; improved; Legal patent; Length; Licensing; Liquid Chromatography; Marketing; Methods; Modification; nano; nanofluidic; Paper; particle; Particle Size; Peptides; Phase; Preparation; pressure; Proteins; public health relevance; Research Personnel; Silicon Dioxide; Small Business Technology Transfer Research; Speed (motion); success; Surface; Technology; Time; Universities; Width

The goal of this STTR Phase II collaboration between Purdue University and bioVidria, Inc. is to commercialize disruptive new technology for protein chromatography and then maximize its impact by serving the fastest growing segment of the pharmaceutical industry: protein drugs. The product is a reversed phase chromatography column that uses silica particles of only 500 nm in diameter, thereby significantly increasing resolution and sensitivity in protein separations. Slip flow is what enables high flow rates with such small particles. The primary market opportunity is the analysis of the heterogeneity of bioengineered drugs based on monoclonal antibodies. These drugs offer high target specificity, e.g., toxicity directed at the tumor. The problem to be solved is that monoclonal antibodies can undergo intra- and inter-molecular disulfide scrambling during storage, which causes immunogenicity in patients. The proposed slip-flow column will uniquely resolve and isolate these scrambled versions, enabling rational design of formulations to reduce immunogenicity. The Phase I research focused on protein and peptide separations in packed capillaries, demonstrating ten-fold narrower zones and a ten-fold flow enhancement from slip flow. The Phase II research will focus on scaling up to develop packed stainless steel columns of 2.1 mm in diameter. The scale-up will allow customers to use these columns with current commercial instruments, which is essential for serving the pharmaceutical industry. The Specific Aims of the Phase II proposal are to 1) optimize the process for packing 500 nm particles into stainless steel columns, 2) develop a scalable process to make bonded phases with negligible silanol activity, and 3) develop separation methods for resolving products of disulfide rearrangement of monoclonal antibodies for all three types of antibody platforms. We enlist three major companies to provide samples: Genentech (IgG1), Pfizer (IgG2) and Eli Lilly (IgG4).

Thesaurus Terms:
Accounting;Adopted;Adverse Drug Effect;Antibodies;Autoimmune Diseases;Base;Biomedical Engineering;Blood Capillaries;Caliber;Capillary;Chromatography;Collaborations;Column Chromatography;Commercialization;Crystal Formation;Design;Development;Diagnostic;Diffusion;Disulfides;Dna Sequence Rearrangement;Drug Formulations;Drug Industry;Goals;Height;Heterogeneity;Humidity;Igg1;Igg2;Igg4;Immunogenicity;Immunoglobulin G;Innovation;Instrument;Length;Malignant Neoplasms;Manufacturer Name;Marketing;Measurement;Methods;Molecular;Monitor;Monoclonal Antibodies;New Technology;Particle;Patients;Peptides;Performance;Pharmaceutical Preparations;Pharmacologic Substance;Phase;Polymerization;Preparation;Procedures;Process;Protein Isoforms;Proteins;Public Health Relevance;Reagent;Relative (Related Person);Reproducibility;Research;Resolution;Reversed Phase Chromatography;Sampling;Scale Up;Self Assembly;Silane;Silanes;Silanol;Silicon Dioxide;Small Business Technology Transfer Research;Specificity;Stainless Steel;Toxic Effect;Tumor;Universities;Water;Width;