SBIR-STTR Award

Biopharmaceuticals are complex molecules manufactured in living cells, and as a result, frequent quality testing is required for development, regulatory approval, and manufacturing. Capillary isoelectric focusing (cIEF) is a widely used analytical technique that provides rapid detection of most protein modifications. Mass spectrometry (MS) measures intrinsic molecular weight, and has the unique capability of identifying the specific nature of a given protein modification. Combining cIEF and MS is a powerful and highly effective approach to determining compositional differences between protein isoforms. However, current methodologies require labor intensive preparative chromatography to isolate sufficient material for the combined cIEF and MS analysis, and so are severely limited in capacity and throughput. To address this issue, Intabio has developed the Blaze™ system, a proprietary protein analytics platform that will be the first commercially available system to seamlessly integrate cIEF with MS analysis. The Blaze system utilizes a novel microfluidic chip design to integrate (1) separation of protein isoforms by isoelectric focusing, (2) imaging of protein isoforms by 280nm absorption for detection and quantitation, and (3) MS sample preparation and delivery by electrospray into an adjacent mass spectrometer to detect and identify each isoform. The Blaze system's integration of these steps drastically reduces the analysis time to only minutes per sample. This SBIR project will advance the development of the Blaze instrument, software and consumable kit, and will establish a consumable manufacturing process to prepare for commercial launch. Phase 1 will focus on advancing the core innovation - the chip and clamshell. The clamshell holds the microfluidic chip in the instrument and acts as the world-to-chip-interface, providing inputs for reagents, samples, pressure and electrodes. In Phase 1, the laser bonding process for the three-layer chip will be optimized to improve yield and chip performance by exploring multiple combinations of bonding parameters. Additionally, prototype designs to integrate the chip and clamshell into a single “cartridge” for improved ease-of-use will be designed and tested. Phase 2 will advance the prototype cartridge to a commercial, disposable version and set up the commercial manufacturing process for the chip cartridge. Commercial instrument and software development will also be completed in Phase 2. Instrument development will address strategies to reduce material cost and integrate an internal autosampler function; software development will refine integration and display of cIEF and MS data, as well as complete cfr21part11 compliance. The innovation and research from this project will advance the fields of protein separation science and expand the microfluidic technology field. This project will also transform biopharmaceutical drug development and manufacturing by enabling more frequent, comprehensive testing. This will result in profound productivity gains, translating into a greater number of higher quality drugs developed in less time and with less cost.  

Public Health Relevance Statement:
Project Narrative The development of the BlazeTM instrument system for real-time protein analytics will address a critical bottleneck in the development and manufacturing of biopharmaceuticals, enabling higher quality drugs to be developed in less time and preventing manufacturing issues that could delay the availability of these important drugs to patients. The Blaze system will accomplish this by being the first commercially available system to integrate three key analytical functions: cIEF separation for protein isoform detection, whole column imaging for quantitation, and ionized electrospray into a back-end mass spectrometer for analysis. This innovation will greatly reduce the time and lab infrastructure required for protein analytics, enabling drug developers to have access to high quality information during the biopharmaceutical development and manufacturing process much sooner and on many more samples than is currently possible.

Project Terms:
absorption; Address; Advanced Development; Affect; Back; base; Biological Products; Blood capillaries; Cells; Charge; ChIP-on-chip; Chromatography; commercialization; Complex; Computer software; cost; Data; Data Display; design; Detection; Development; Device or Instrument Development; drug development; drug quality; effective therapy; Electrodes; Generations; graphical user interface; Heterogeneity; Image; improved; innovation; instrument; instrumentation; Ions; Isoelectric Focusing; Lasers; lens; manufacturing process; mass spectrometer; Mass Spectrum Analysis; Measures; Methodology; Microfluidic Microchips; microfluidic technology; Microfluidics; Modification; Molecular Weight; Monoclonal Antibodies; Nature; novel; Optics; Patients; Performance; Pharmaceutical Preparations; Phase; Positioning Attribute; Post-Translational Protein Processing; Preparation; pressure; prevent; Process; Production; Productivity; Protein Isoforms; Proteins; prototype; rapid detection; Reagent; Real-Time Systems; Research Infrastructure; Research Project Grants; Risk; Sampling; Science; Small Business Innovation Research Grant; software development; System; Systems Integration; Techniques; Testing; Time; tool; Toxic effect; Translating; Vertebral column

Biopharmaceuticals are complex molecules manufactured in living cells, and as a result, frequent quality testing is required for development, regulatory approval, and manufacturing. Capillary isoelectric focusing (cIEF) is a widely used analytical technique that provides rapid detection of most protein modifications. Mass spectrometry (MS) measures intrinsic molecular weight, and has the unique capability of identifying the specific nature of a given protein modification. Combining cIEF and MS is a powerful and highly effective approach to determining compositional differences between protein isoforms. However, current methodologies require labor intensive preparative chromatography to isolate sufficient material for the combined cIEF and MS analysis, and so are severely limited in capacity and throughput. To address this issue, Intabio has developed the Blaze™ system, a proprietary protein analytics platform that will be the first commercially available system to seamlessly integrate cIEF with MS analysis. The Blaze system utilizes a novel microfluidic chip design to integrate (1) separation of protein isoforms by isoelectric focusing, (2) imaging of protein isoforms by 280nm absorption for detection and quantitation, and (3) MS sample preparation and delivery by electrospray into an adjacent mass spectrometer to detect and identify each isoform. The Blaze system's integration of these steps drastically reduces the analysis time to only minutes per sample. This SBIR project will advance the development of the Blaze instrument, software and consumable kit, and will establish a consumable manufacturing process to prepare for commercial launch. Phase 1 will focus on advancing the core innovation - the chip and clamshell. The clamshell holds the microfluidic chip in the instrument and acts as the world-to-chip-interface, providing inputs for reagents, samples, pressure and electrodes. In Phase 1, the laser bonding process for the three-layer chip will be optimized to improve yield and chip performance by exploring multiple combinations of bonding parameters. Additionally, prototype designs to integrate the chip and clamshell into a single “cartridge” for improved ease-of-use will be designed and tested. Phase 2 will advance the prototype cartridge to a commercial, disposable version and set up the commercial manufacturing process for the chip cartridge. Commercial instrument and software development will also be completed in Phase 2. Instrument development will address strategies to reduce material cost and integrate an internal autosampler function; software development will refine integration and display of cIEF and MS data, as well as complete cfr21part11 compliance. The innovation and research from this project will advance the fields of protein separation science and expand the microfluidic technology field. This project will also transform biopharmaceutical drug development and manufacturing by enabling more frequent, comprehensive testing. This will result in profound productivity gains, translating into a greater number of higher quality drugs developed in less time and with less cost.  

Public Health Relevance Statement:
Project Narrative The development of the Blaze™ instrument system for real-time protein analytics will address a critical bottleneck in the development and manufacturing of biopharmaceuticals, enabling higher quality drugs to be developed in less time and preventing manufacturing issues that could delay the availability of these important drugs to patients. The Blaze system will accomplish this by being the first commercially available system to integrate three key analytical functions: cIEF separation for protein isoform detection, whole column imaging for quantitation, and ionized electrospray into a back-end mass spectrometer for analysis. This innovation will greatly reduce the time and lab infrastructure required for protein analytics, enabling drug developers to have access to high quality information during the biopharmaceutical development and manufacturing process much sooner and on many more samples than is currently possible.

NIH Spending Category:
Bioengineering; Biotechnology; Networking and Information Technology R&D (NITRD)

Project Terms:
absorption; Address; Advanced Development; Affect; Back; base; Biological Products; Blood capillaries; Cells; Charge; ChIP-on-chip; Chromatography; commercialization; Complex; Computer software; cost; Data; Data Display; design; Detection; Development; Device or Instrument Development; drug development; drug quality; effective therapy; Electrodes; Generations; graphical user interface; Heterogeneity; Image; improved; Infrastructure; innovation; instrument; instrumentation; Ions; Isoelectric Focusing; Lasers; lens; manufacturing process; mass spectrometer; Mass Spectrum Analysis; Measures; Methodology; Microfluidic Microchips; microfluidic technology; Microfluidics; Modification; Molecular Weight; Monoclonal Antibodies; Nature; novel; Optics; Patients; Performance; Pharmaceutical Preparations; Phase; Positioning Attribute; Post-Translational Protein Processing; Preparation; pressure; prevent; Process; Production; Productivity; Protein Isoforms; Proteins; prototype; rapid detection; Reagent; Real-Time Systems; Research Project Grants; Risk; Sampling; Science; Small Business Innovation Research Grant; software development; System; Systems Integration; Techniques; Testing; Time; tool; Toxic effect; Translating; Vertebral column