SBIR-STTR Award

?The very forces critical to creating the three dimensional structure of proteins are also responsible for unwanted and often dangerous protein aggregates in protein-­based therapeutics. The presence of such aggregates can mean the discontinuation of potentially critical therapeutic products. The FDA has advised in its 2014 Guidance for Industry that ''It is critical for manufacturers of therapeutic protein products to minimize protein aggregation to the extent possible'', while acknowledging that the technology does not currently exist to accomplish this goal, particularly in the critical range from 100 nm to 10 microns. Protein aggregation occurs at almost every stage in the biopharmaceutical pipeline, from development, formulation, and manufacturing to storage and point of use. Thus, there is a pressing and as-yet unmet need for instrumentation that can assess the size distribution and morphology of protein aggregates in this size range. This SBIR Phase I program will meet this need by developing an instrument, using holographic video microscopy to detect and characterize protein aggregates in pharmaceutical formulations. This transformative technology, originally invented to characterize the properties of spherical colloids and emulsion droplets, can measure single clusters rapidly enough to be used for in-line characterization of pharmaceutical samples, and offers unprecedented and independently verified precision and accuracy. The proposed work will modify existing patented instrumentation and software for application to the measurement of protein aggregates in model systems, will verify the instrument's ability to distinguish protein clusters from industrially relevant contaminants, and will demonstrate the ability to monitor protein-clustering behavior under conditions that enhance or inhibit aggregation. Holographic characterization of protein aggregation will provide the detailed data needed for product development, process control and quality assurance in the biopharmaceutical industry and will result in increased efficiency, safety and cost savings in the promising and rapidly growing area of protein-based therapeutics.

Public Health Relevance Statement:


Public Health Relevance:
The tendency for proteins to aggregate is a critical barrier to developing and manufacturing biopharmaceuticals for targeting a wide array of diseases, largely because there are no fast, effective and reliable methods to detect and characterize these clusters. This proposed SBIR Phase I program will develop an instrument that uses holographic video microscopy to meet this need, resulting in increased efficiency, safety, and cost savings in the promising and rapidly growing area of protein-based therapeutics. Holographic characterization of protein aggregates will be validated by measuring two model proteins, bovine serum albumin and bovine insulin, under a variety of conditions designed to modify the aggregation behavior of these proteins.

NIH Spending Category:
Bioengineering; Biotechnology

Project Terms:
analytical method; Anaphylaxis; Area; base; Behavior; Biological Models; Biological Products; Bovine Serum Albumin; Buffers; Categories; Cattle; Chemicals; Clinical; Colloids; Computer software; Cost Savings; cytokine; Data; design; Development; Disease; Emulsions; Environmental Risk Factor; Event; Formulation; Goals; Growth; Habits; Health; Immune response; Individual; Industry; instrument; instrumentation; Insulin; Latex Particles; Legal patent; Manufacturer Name; Measurement; Measures; Mediating; meetings; Methods; Modeling; Monitor; Morphology; nanometer; Optics; particle; Pharmacologic Substance; Phase; Process; product development; programs; Property; protein aggregate; protein aggregation; Proteins; Protocols documentation; quality assurance; research and development; Safety; Sampling; Silicon Dioxide; Silicone Oils; Small Business Innovation Research Grant; Staging; Syndrome; System; Techniques; Technology; Temperature; Therapeutic; therapeutic protein; three dimensional structure; Video Microscopy; Work

The very forces critical to creating the three-dimensional structure of proteins are also responsible for unwanted and often dangerous protein aggregates in protein- based therapeutics. Protein aggregation occurs at every stage in the biopharmaceutical pipeline, from development, formulation, and manufacturing to shipping, storage and point of use. The FDA has advised in its 2014 Guidance for Industry that “It is critical for manufacturers of therapeutic protein products to minimize protein aggregation to the extent possible”, while acknowledging that the technology does not currently exist to accomplish this goal, particularly in the critical sub-visible range from 100 nm to 10 microns. Thus, there is a pressing and as-yet unmet need for instrumentation that can assess the size distribution and morphology of protein aggregates in this size range. This SBIR Phase II program will meet this need by developing an instrument that uses holographic video microscopy to characterize protein aggregates rapidly and reliably enough for in-line monitoring of pharmaceutical formulations. The proof-of-principle studies performed under the Phase I grant demonstrated the accuracy of holographic characterization in measuring the size and optical properties of protein aggregates, and reliably distinguishing them from silicone oil droplets or other contaminants. The Phase II work will harness these patented capabilities for automated high- throughput measurement of real-world samples. Hardware and software innovations will be required to ensure accuracy and efficacy in high-speed characterization, and to further increase the sophistication and reliability of particle differentiation even when different components are the same size. This program will also elucidate the influence of increased flow speed on the growth rate and morphology of protein aggregates. Holographic characterization of protein aggregation will provide the detailed data needed for product development, process control and quality assurance in the biopharmaceutical industry and will result in increased efficiency, safety and cost savings in the promising and rapidly growing area of protein-based therapeutics.

Public Health Relevance Statement:
Project Narrative The tendency for proteins to aggregate is a critical barrier to developing and manufacturing biopharmaceuticals for targeting a wide array of diseases, largely because there are no fast, effective and reliable methods to detect and characterize these clusters. This proposed SBIR Phase II program will develop an instrument that uses holographic video microscopy to meet this need, resulting in increased efficiency, safety, and cost savings in the promising and rapidly growing area of protein-based therapeutics. Holographic characterization of protein aggregates will be adapted and tested for use in the biopharmaceutical industry by simultaneously measuring the concentrations of similarly sized different components in real-world samples, with high-throughput microfluidic sample handling.

Project Terms:
Accounting; Affect; Anaphylaxis; Area; base; Biological Products; cell dimension; Cells; Clinical; Complex; Computer software; Cost Savings; cross reactivity; cytokine; Dangerousness; Data; design; Development; Dimensions; Disease; Drug Industry; Ensure; Event; experimental study; Formulation; Goals; Grant; Growth; Immune response; Industry; Injection of therapeutic agent; innovation; instrument; instrumentation; Legal patent; Manufacturer Name; Measurement; Measures; Mediating; Methods; Microfluidics; Modeling; Monitor; Morphology; nanometer; Optics; particle; Pharmacologic Substance; Phase; pressure; Process; product development; programs; Property; protein aggregate; protein aggregation; protein structure; Proteins; prototype; quality assurance; Refractive Indices; Safety; Sampling; Shipping; Silicone Oils; Small Business Innovation Research Grant; Speed; Suspensions; Syndrome; System; Technology; Testing; Therapeutic; therapeutic protein; three dimensional structure; Treatment Efficacy; Video Microscopy; Work