SBIR-STTR Award

Nanoparticles are ubiquitous yet notoriously difficult to detect and measure-practical technologies for their characterization are effectively restricted to particles larger than about 500 nanometers in diameter. This limitation is acutely felt in the drug development industry: Because aggregate drug particles in parenteral formulations elicit a severe immune response in some recipients, their detection is a major safety concern for manufacturers and patients. Millions of these critical tests are performed each year across the pharmaceutical industry, and the lack of effective particle sizing technology significantly increases the time and cost of these tests: a 1000-fold reduction in both time and reagents required for each test would be realized if molecular aggregates could be reliably detected at diameters of 50 nanometers rather than 500 nanometers. Spectradyne will address this unmet need by commercializing an innovative particle sizing technology that completely changes the way submicron aggregates are detected and measured in the pharmaceutical industry. The Spectradyne instrument will be capable of 1) robust detection of individual particles as small as 20 nanometers in diameter, more than ten times smaller than existing commercially available methods, with 2) size discrimination better than 5%, enabling resolution of polydisperse samples and 3) detection rates higher than 100,000 particles per second, (1000-fold faster than other commercially available methods). The platform will comprise single-use consumable nanofluidic cartridges and a compact, benchtop instrument capable of controlling flow in the cartridge and performing electrical readout of particles as they flow through the sensor. To accomplish these goals three specific aims will be met. In Aim 1, reliable and repeatable manufacture of the consumable nanofluidic cartridges will be demonstrated-successful completion of this aim will define a viable path to mass manufacture of these components, a critical requirement for the success of the technology. In Aim 2, an instrument prototype will be developed that is capable of automated control of fluid flow in the cartridge and electrical readout. Success in Aims 1 and 2 will produce a reliable control system with which to systematically evaluate the performance of the technology. Lastly, in Aim 3 the key performance metrics of the system will be characterized: limit of detection, linear dynamic range, and measurement accuracy and precision. The fully characterized instrument prototype will then be validated for use in drug development applications by analyzing authentic formulations provided by our collaborators in the pharmaceutical industry. This technology will significantly reduce the time, money and materials required for the development of new drug formulations, in particular the detection of particle aggregation, and reduce the overall time and cost of drug development.

Public Health Relevance Statement:


Public Health Relevance:
All parenteral drug formulations must be continually tested for the formation of aggregate drug particles, which are known to illicit severe immune responses in some recipients and which can require weeks or months to grow to a size large enough to be detected using currently available methods (about 500 nanometers in diameter). This project will develop an innovative new technology that integrates state-of-the-art nanofluidic technology with rapid electronic sensing in a practical, compact instrument to reduce the minimum detectable particle size to below 50 nanometers. The ability to detect particles in this size range will enable 1000-fold reductions in both the time required to detect aggregation and the quantity of sample required for analysis, thus greatly reducing the time and cost in this step of drug development.

Project Terms:
Address; Automation; base; Caliber; commercialization; constriction; Cosmetics; cost; Detection; Development; Dimensions; Discrimination (Psychology); Drug Delivery Systems; drug development; Drug Formulations; Drug Industry; Electric Conductivity; Electronics; Environment; Feedback; fluid flow; Food; Foundations; Future; Geometry; Goals; Health; Immune response; improved; Indium; Individual; Industry; industry partner; innovation; instrument; instrumentation; Letters; Liquid substance; Manuals; Manufacturer Name; manufacturing process; Measurement; Measures; meetings; member; Methods; Metric System; Molds; Molecular; nanofluidic; nanometer; nanoparticle; nanopattern; nanoscale; Nanotechnology; new technology; operation; particle; Particle Size; Patients; Performance; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Play; Polystyrenes; pressure; Process; programs; prototype; public health relevance; Reagent; Resolution; response; Safety; Sampling; sensor; Signal Transduction; Small Business Innovation Research Grant; submicron; success; System; Technology; Testing; Time; tool; Translating; United States National Institutes of Health; Variant; visual feedback

Nanoscale particles in serum play critical roles in biological processes, including intercellular signaling, immune response, and the pathophysiology of important diseases such as cancer. One such disease is Chronic Kidney Disease-Mineral and Bone Disorder (CKD-MBD)-a clinical entity that is observed in a majority of the ~20 million Americans with CKD. CKD-MBD promotes vascular calcification that leads to progressive cardiovascular dysfunction, and significantly contributes to the exceedingly high rate of cardiovascular morbidity/mortality observed in this population. In CKD-MBD, the formation of soluble nanoscale calciprotein particles (CPPs) in serum mitigates the toxicity of hyperphosphatemia by sequestering calcium phosphate crystals and preventing their precipitation in the vasculature. Exciting recent discoveries have shown that CPP concentrations increase with CKD progression, and are not routinely observed in patients with normal kidney function. CPPs may thus represent a novel biomarker for identifying patients at high risk for progressive vascular calcification prior to the onset of irreversible mineralization f the vasculature. However, characterization of CPPs to date has been hindered by an inability to efficiently measure these nanoparticles in clinical samples. Spectradyne has developed a powerful platform instrument, the nCS1, which accurately measures the size and concentration of polydisperse nanoparticles in complex matrices such as serum. The technology is ideally suited to characterize biological particles as indicators of health and disease. Spectradyne proposes to collaborate with researchers at the University of Kansas Medical Center (KUMC) to build a version of this instrument for quantification of CPPs in serum. Together the combined team will investigate the potential of using the size distribution of particles in serum samples to predict the risk for vascular calcification in mice and humans, with the long-term goal of improving diagnosis and prevention of this condition. To accomplish these goals, two specific aims will be met. In Aim 1, the instrument will be configured for measurements of serum nanoparticles: Sample preparation, the instrument itself, the analysis cartridge, and the data analysis software will all be optimized for measuring nanoparticles in serum. Cartridge production will also be increased for the study. Aim 2 will evaluate the ability of the reconfigure instrument to measure CPPs in sera from humans and mice at high risk for vascular calcification. To achieve this we will: A) Quantify serum CPP concentrations and associated vascular calcification in two murine models; and B) Compare serum CPP levels in CKD patients and healthy controls, and associate with traditional mineral metabolism markers. Completion of this work will yield a user-friendly bench top instrument capable of rapid, high-resolution particl analysis in serum samples. Moreover, the utility of the instrument for the quantification of serum CPPs will be evaluated, and the role of this novel biomarker in the pathophysiology of vascular calcification determined.

Public Health Relevance Statement:


Public Health Relevance:
Nanoscale particles in serum play a significant role in a broad set of biological processes that contribute to the development of important human diseases. Patients with decreased kidney function exhibit a disproportionate burden of calcium-phosphate buildup in their arteries (vascular calcification), which ultimately results in these patients suffering from a much higher rate of heart failure and death when compared to the general population. Recent evidence suggests that patients with decreased kidney function exhibit elevated blood levels of microscopic calcium-phosphate crystals called calciproteins that may promote vascular calcification formation. This project will further develop a revolutionary instrument for detecting calciproteins and other nanoparticles directly in blood, and thoroughly evaluate the utility of these particles as a diagnostic tool to promote the earlier diagnosis and improved treatment of vascular calcification in patients with kidney disease.

NIH Spending Category:
Bioengineering; Cardiovascular; Kidney Disease; Nanotechnology; Prevention

Project Terms:
Accounting; American; Animals; Appearance; Arteries; Automation; Automobile Driving; Basic Science; Biological; Biological Markers; Biological Process; Blood; Blood Circulation; Bone Diseases; calcification; calcium phosphate; Caliber; Cardiac; Cardiovascular Diseases; Cardiovascular Pathology; cardiovascular risk factor; Cardiovascular system; Cessation of life; Chronic Kidney Failure; Clinical; Clinical Research; clinical risk; Complex; Computer software; cytotoxic; Data; Data Analyses; Detection; Development; Device or Instrument Development; Diagnosis; Diagnostic; Diet; Disease; Disease Progression; Early Diagnosis; End stage renal failure; Ensure; Event; Exhibits; FGF3 gene; Functional disorder; Future; General Population; Goals; Health; Heart failure; high risk; Hospitalization; Human; human disease; Immune response; improved; inorganic phosphate; instrument; Kansas; Kidney Diseases; Malignant Neoplasms; Measurement; Measures; Medical center; meetings; Metabolism; Methods; Microscopic; mineralization; Minerals; Modeling; Morbidity - disease rate; Mortality Vital Statistics; Mus; nanoparticle; nanoscale; novel; novel diagnostics; particle; Pathologic Processes; Patients; Physiological Processes; Play; Population; Precipitation; Preparation; prevent; Prevention; Production; Protein Binding; Protocols documentation; public health relevance; Reference Standards; Renal function; Research Personnel; research study; Resolution; Risk; Risk Factors; Rodent; Role; Sampling; Serum; Signal Transduction; Specimen; Staging; success; Technology; Temperature; tool; Toxic effect; Treatment Efficacy; Universities; user-friendly; Validation; Vascular calcification; Work