Agency: Department of Defense | Branch: Defense Advanced Research Projects Agency | Program: STTR | Phase: Phase I | Award Amount: 99.35K | Year: 2012
With the notable exception of glucose, most metabolic biomarkers cannot be accurately, and practically measured over long periods, in vivo and in real-time. A primary reason is stable, selective enzymes suitable for biosensors are not available. Biosensors could revolutionize acute and long-term monitoring in the development, clinical, first responder and battlefield arenas. Pinnacle has developed intellectual property (patent PCT/US11/51193) that provides a general solution for the use of cofactors to enhance the biosensors sensitivity. We have also discovered that fusion proteins can dramatically enhance the stability of oxidases without compromising catalytic activity. Using high throughput screening in a cell-free system, directed evolution and large scale protein production in E.coli, we propose to develop a platform methodology for the optimization of existing (e.g. histamine), and the creation of new, (e.g.cortisol) oxidases for biosensors. Pinnacle Technology manufactures, sells and warranties biosensors for the detection of glucose, glutamate, lactate, and alcohol in animals. With the Richter and Gao laboratories at the University of Kansas we have successfully cloned, expressed, purified and characterized oxidases for biosensors. The enzymes produced will be available for direct sale, or as biosensors. World-wide total biosensor sales are expected to reach $14 billion by 2016.
Agency: Department of Defense | Branch: Defense Advanced Research Projects Agency | Program: STTR | Phase: Phase II | Award Amount: 723.76K | Year: 2014
Point of care (POC) diagnostic devices are a well established market but have significant growth opportunities. Stable off-the-shelf biological recognition elements available in a form that can be readily incorporated into POC devices will be essential for wide spread use and deployment. Other than glucose oxidase, stable active oxidase enzymes are not available. Pinnacle has developed intellectual property (patent PCT/US11/51193) that provides a general solution for the use of cofactors to maximize the activity of oxidase enzymes used in diagnostic devices. We have also discovered that fusion proteins can dramatically enhance the stability of oxidases without compromising catalytic activity. Using high throughput screening of fusion proteins, directed evolution and large scale protein production in E.coli, we are developing a platform methodology for the optimization of existing (e.g. histamine, lactate), and the creation of new (e.g.cortisol) oxidases. Pinnacle Technology manufactures, sells and warranties biosensors for the detection of glucose, glutamate, lactate, and alcohol in animals. With the Richter and Gao laboratories at the University of Kansas, we have successfully cloned, expressed, purified and characterized oxidases for biosensors. The enzymes produced will be available for direct sale, or as biosensors. World-wide total biosensor sales are expected to reach $14 billion by 2016.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 150.01K | Year: 2012
DESCRIPTION (provided by applicant): Degeneration of sleep quality with age is a well-documented phenomenon. While much is known regarding the characterization of this decline, remarkably little of the physiology or genetic underpinnings are understood. Age-related sleep changes in rodents (mice + rats) are remarkably similar to those seen in human populations and as such these animal models are frequently used to study the aging process. However, the current state-of-the-art technology necessary for measuring sleep in rodents requires surgical implantation of EEG and electromyograph (EMG) probes. Drawbacks to this process include the need for an invasive and time-consuming animal surgery paired with a prolonged recovery period. The time component is extremely slow as the animal must recover from surgery and acclimate to the recording chamber and tether before normal sleep can be evaluated. Additionally, biofouling of the implanted screws and decay of the chronic implant alters signal quality over time and restricts long-term recordings to only a few continuous months. Labor expenses include the need to hire an expert technician to run the equipment, undertake surgery and evaluate sleep data. To overcome these difficulties we plan to use accelerated video hardware, 3-D video monitoring and a deep red grid (invisible to the rodent) to measure sleep state. In Phase I of this application, Pinnacle Technology Inc and the University of Pennsylvania Center for Sleep and Respiratory Neurobiology will create a workingprototype (software, electronics, and hardware) for non-invasive detection of sleep state in rodents. Specifically, this system aims to be the first commercial device designed to record and automatically analyze wake, NREM and REM sleep in rodents using non-surgical, non-invasive techniques. A unique benefit to this design is that it will be capable of continuously recording sleep-wake architecture from the same animal across the lifespan. The system, once validated, can easily be used as a large-scale screening tool to rapidly screen hundreds of animals. Commercial applications of such a device are already being sought by major pharmaceutical companies as they explore new potential gene products for targeted therapy. PUBLIC HEALTH RELEVANCE: It is estimated that at least 40 million Americans suffer each year from chronic, long-term sleep disorders while an additional 20 million experience occasional sleeping problems. As people age, sleeping disorder are more prevalent; more than 50 percent of peopleolder than 64 have a sleep disorder. Economically, poor sleep accounts for an estimated 16 billion in medical costs each year and the indirect costs of lost productivity and other factors are known to be much higher.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 149.91K | Year: 2012
DESCRIPTION (provided by applicant): Tobacco-related deaths cost the United States approximately 200 billion each year. Nicotine is singularly responsible for the dependence-forming properties of tobacco smoking and, once introduced to the body, rapidly affects the brain within 15 seconds. Breaking nicotine addiction is challenging and relapse rates remain high. The extremely rapid timescale of nicotine action renders existing techniques for studying nicotine accumulation in the brain (e.g., microdialysis,PET, radioimmunoassay) ill-suited for long-term addiction studies in freely moving animals. Biosensors are a proven technology for monitoring real-time changes in CNS neurochemical concentrations. The most critical component of a biosensor is the enzyme used as the biorecognition element, and no aspect of a biosensor's final design is as vital as a properly folded enzyme with sufficient activity and stability profiles. To date, no nicotine-specific enzyme has been reported. A primary goal of this Phase I SBIR proposal is to begin the transformation of an existing oxidase enzyme's kcat, stability at 37oC, Km, and Tm into an enzyme capable of the specific detection of nicotine - a process that will be completed during Phase II. We will combine structure-guided design with directed protein evolution (including a suitable selection process) to hone and optimize a new nicotine oxidase enzyme suitable as the foundation of a nicotine biosensor. The resulting nicotine biosensor promises to impact nicotine addictionstudies by allowing real-time recordings of CNS nicotine concentrations in freely moving animals for up to one week. Furthermore, the nicotine biosensor could be used in conjunction with other biosensors, which will allow for the simultaneous monitoring ofchanges in nicotine concentration in tandem with other important CNS analytes (i.e., glucose, glutamate, lactate) or addictive compounds (i.e., ethanol). This approach promises to reveal new insights into nicotine distribution, dynamics and flux. Furthermore, a nicotine biosensor should find use as a screening tool for the development of new pharmacologic agents designed to assist in smoking cessation and inhibit relapse. The completion of both Phase I and Phase II components will provide two important innovations to the scientific community: 1) A nicotine biosensor suitable for long-term (i.e., up to one week) addiction studies that provides second-by-second changes of nicotine concentration in the CNS. 2) A refined approach for the development of new biosensors that target analytes important for addiction and for which no oxidase enzyme currently exists (i.e., cocaine and caffeine). PUBLIC HEALTH RELEVANCE: It is estimated that 50% of regular smokers die due to smoking-related complications. In addition, nicotine addiction associated with cigarette smoking takes a tremendous economic toll, costing the United States nearly 200 billion annually, including 97 billion in lost productivity and 96 billion in health care expenditures. This projec allows for a better understanding of nicotine addiction, which could advance smoking cessation research and potentially prevent 450,000 smoking-related deaths per year in the United States.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 144.31K | Year: 2014
ABSTRACT Age-related sleep problems such as advanced sleep phase disorder (ASPD) are estimated to affect at least 1% of middle-aged adults and increase in prevalence with age. While the detrimental effects of sleep disruption with aging are well characterized, detailed insights into the molecular and physiological mechanisms underlying these sleep changes are greatly lacking. Optogenetics harnesses a combination of genetic and optical methods to directly control neuronal events in specific cells of the central nervous system. Recent studies have confirmed that control of both wakefulness and slow-wave-sleep are possible using optogenetic methods. These methods can be used to provide an unprecedented understanding of cortical activity in aging. The optogenetics field is maturing and there are numerous commercial sources for optogenetic components; however, the technique requires a multidisciplinary skill set including chemistry, optics, physiology, electronics, mechanics, software, and systems analysis. To da
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase II | Award Amount: 998.76K | Year: 2015
DESCRIPTION provided by applicant Abstract In this Phase II SBIR we will commercialize tissue implantable microbiosensors to measure glucose lactate and oxygen for use in freely moving animals We will also develop and commercialize a sensor array composed of a glucose biosensor a lactate biosensor and an oxygen sensor Biosensors provide high temporal resolution and low analyte consumption when compared to microdialysis Current state of the art biosensors O D m have proven value in the measurement of a range of analytes in the brains of freely moving rodents Reduction of biosensor diameter will result in considerably less damage to brain tissue and improved anatomical accuracy over current microdialysis and biosensor techniques Pinnacle will lead this proposal and work in conjunction with an interdisciplinary consortium of three leading scientists at the University of Kansas Professor Shenqiang Ren possesses extensive experience in materials science and nanofabrication techniques Professor Mark Richter is skilled in protein engineering and Professor George Wilson brings over years of experience in biosensor development to the project The facilities and equipment available at Pinnacle and the various University of Kansas laboratories will provide the resources required to successfully complete this project Innovative aspects of this proposal include new technologies in biosensor manufacturing such as nanoetching and electrophoretically manipulated enzyme immobilized nanoparticles and the ability to routinely and precisely deposit picoliters of substrate into a defined well In Phase I our team made several significant advancements in the development of a prototype m glucose microbiosensor We produced functional prototypes that measured glucose in a linear fashion to at least mM rejected ascorbate and urate and efficiently recycled O at the electrode surface These preliminary results form the foundation for the Phase II proposal These implantable m microbiosensors have the potential to significantly increase the understanding of the mechanisms behind drug neuron interactions This will profoundly change the drug discovery landscape and improve drug development efficiencies for pharmaceutical and biotech companies In addition increased spatial resolution and decreased inflammation responses will allow researchers to identify new neural processes leading to new understandings approaches and solutions for common maladies Biosensor sales in FY were $ billion with of the sales due to human glucose sensors Pinnacle is an established manufacturer of biosensors and is well positioned to introduce this new class of biosensors to a broad market PUBLIC HEALTH RELEVANCE In the United States over disorders of the brain and nervous system result in more hospitalizations than for any other disease group including heart disease and cancer The quality of life and economic cost of brain and nervous system related disorders is staggering These disorders disrupt the lives of more than million Americans each year and costs exceed $ billion
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase II | Award Amount: 923.37K | Year: 2015
DESCRIPTION provided by applicant The goal of this project is to design and commercialize tethered and wireless turn key optogenetics and electrophysiological neurotransmitter behavior measurement systems for use in mice and rats Optogenetics harnesses a combination of genetic and optical methods to directly control neuronal events in specific cells of the central nervous system These methods are broadly applicable but can be specifically used to provide an unprecedented understanding of cortical activity and aging The optogenetics field is maturing and there are numerous commercial sources for optogenetic components however the technique requires a multidisciplinary skill set including chemistry optics physiology electronics mechanics software and systems analysis To date any single experiment requires a system designed from individual component parts Many researchers also have existing equipment that they desire to incorporate into a full optogenetics system This may include lasers cameras and potentially behavioral hardware and software platforms In these situations a digital timing protocol TTL is often used to maintain synchronization but there are subtleties device latency etc to this approach that are often overlooked The proposed system will be capable of delivering multiple selectable wavelengths of light to one or more specific brain regions while simultaneously recording electrical signals neurotransmitters and behavior in rodents throughout the lifespan of the animal All synchronization between the electrophysiological mechanical and visual inputs and optical and stimulus outputs will be precisely controlled via a master timing digital input output platform as well as sophisticated software timing techniques The LED fiber probe connects via a simple electrical connection to a headstage This removes the need for fiber optic rotary joints and enables precise control of the amount of light delivered When completed this system will significantly improve scientific knowledge by providing a turn key solution for researchers from multiple fields to seamlessly integrate optogenetic control alongside traditional pharmaceutical aging and other studies PUBLIC HEALTH RELEVANCE In the United States over disorders of the brain and nervous system result in more hospitalizations than any other disease group including heart disease and cancer Twenty five percent of all years of life lost to disability and premature mortality are due to mental health disorders The overall cost of serious mental illness is estimated to be $ billion a year The widespread use of optogenetics techniques could broadly impact the search for mitigations and cures
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 149.99K | Year: 2012
DESCRIPTION (provided by applicant): The aim of this application is to develop, prototype, validate, manufacture and commercialize a new, generalized microbiosensor designed for non-human use and based on functionalized nanoparticles. The microbiosensors produced will enable in vivo monitoring of a wide range of non-electroactive, neurologically relevant analytes, including, but not limited to, glucose, glutamate, lactate, choline, histamine, D-serine, and ethanol. The design can be extended to multiple conjoined capillaries to provide an array of biosensors for the simultaneous monitoring of multiple analytes, with high temporal and spatial resolution, which causes minimal damage to the surrounding tissue. The specific aim for Phase I is to demonstrate feasibility by prototyping and characterizing a tissue implantable enzyme functionalized nanoparticle microbiosensor that is suitable for in vivo brain measurements and extensible to multianalyte microbiosensor arrays. The proposed microbiosensor will allow researchers to target smaller subregions of the brain that would be damaged by currently available probes. This, in turn, will lead to a better understanding of function and assist in quantifying drug response. The proposed microbiosensor design can also beadapted to the simultaneous monitoring of multiple analytes within a single region of space. Turn-key tools that allow an enhanced level of specificity and granularity will be widely useful in research on the causes, prevention, diagnosis, and treatment ofneurological disorders, and this will lead to new discoveries. Pinnacle will lead this application and work in conjunction with an interdisciplinary consortium of three leading scientists at the University of Kansas. Professor Judy Wu possesses extensiveexperience in materials science and nanofabrication techniques, Professor Mark Richter is skilled in protein engineering, and Professor George Wilson brings over 30 years of experience in biosensor development to the project. The facilities and equipment available at Pinnacle, and the various University of Kansas laboratories, will provide the resources required to successfully complete this project. Innovative aspects of this application include the use of active enzyme functionalized nanoparticles as thesensing element, and the controlled electrophoretic deposition of these nanoparticles. Also novel is a design that maximizes oxygen reuse and hydrogen peroxide detection efficiency. This will lead to linear microbiosensors with high sensitivity for the analyte being monitored. Extensive preliminary results are presented in the application. These results are the foundation for this Phase I effort. Overall, worldwide biosensor sales in FY2009 were 6.9 billion with 31 percent of the sales due to human glucose sensors. Pinnacle is an established manufacturer of biosensors and is well positioned to introduce this new class of biosensors to a broad pre-clinical market. PUBLIC HEALTH RELEVANCE: In the United States, over 1000 disorders of the brain and nervous system result in more hospitalizations than for any other disease group including heart disease and cancer. The quality of life, and economic, cost of brain and nervous system related disorders, is staggering. These disorders disrupt the lives of more than 50 million Americans each year and costs exceed 400 billion.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase II | Award Amount: 866.98K | Year: 2011
DESCRIPTION (provided by applicant): Within the last seven years, Drosophila melanogaster or common fruit fly has gained an important niche in becoming an invertebrate model organism for exploring genetic and proteomic mechanisms underlying sleep. While agreat deal of progress has been made in the field of Drosophila sleep research using available tools, they are limited for the observation, recording and analysis of sleep (immobility) periods in the fruit fly. Additionally, no existing system is capable of individually sleep-depriving flies on an as- needed basis. This proposal outlines a system explicitly designed to measure small amounts of motion in the Drosophila and, as such, more accurately estimate sleep than any available system. In conjunction with this detection device we also propose the design of a mechanized system capable of automatically sleep depriving individual flies through a fully-programmable computerized interface. The system will also provide a flexible behavioral platform for the controlled introduction of modalities based on light, temperature, scent, sound, etc. Phase I of this proposal successfully demonstrated a device to record and analyze motion patterns and sleep as well as enforce wakefulness in individually housed flies. Phase II will build this prototype device into an automated, scalable, modular system capable of monitoring and sleep depriving 160+ flies housed within a commercial incubator. Accomplishment of this project will involve a collaborative effort between PinnacleTechnology and the world-renowned Drosophila research team at The University of Pennsylvania. Prototyping and design will continue to be undertaken by Pinnacle Technology while testing and refinement of the system will be accomplished by the University ofPennsylvania team. The combination of these two features specifically designed to record and automatically analyzes sleep in the fly, will provide a new and powerful tool for Drosophila researchers to explore the basic genetics underlying sleep. Technological innovations of this system will include enhanced video recognition software, mechanisms to observe multiple flies with a single video camera and the technology to selectively deprive individual flies of sleep. Control of individual fly sleep deprivation and real-time visualization of data will be available though a web-based control system. After completion, the system will be validated by testing with known sleep mutations and validated by a trained human observer. Commercial applications of such a device are already being sought by major pharmaceutical companies as they explore new potential gene products for targeted therapy. PUBLIC HEALTH RELEVANCE: It is estimated that at least 40 million Americans suffer each year from chronic, long-term sleep disorders while an additional 20 million experience occasional sleeping problems. Economically, poor sleep accounts for an estimated 16 billion in medical costs each year and the indirect costs of lost productivity and other factors are known to be much higher. While the links between inadequate sleep and poor health and productivity are becoming clearer, there is a dire need to understand the basic causes and biological need for sleep.
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase I | Award Amount: 153.96K | Year: 2016
DESCRIPTION provided by applicant As the most important inhibitory neurotransmitter in the brain a detailed understanding of the implications of gamma aminobutyric acid GABA release remains elusive The measurement of GABA concentrations is a difficult process Microdialysis is the current standard for GABA sampling in the brains of freely moving animals but suffers from low temporal resolution and the need for labor intensive analysis methods By contrast the direct sensing of GABA by modalities including biosensors provides second by second temporal resolution without the need for additional post analysis However biosensors and other monitoring devices require an enzyme to process the analyte of interest The state of the art for the enzymatic conversion of GABA into a transducible signal is the sequential activity of three separate enzymes or antibodies entrapped within nanoparticles For CNS and systemic GABA sensing applications a single GABA oxidase enzyme is necessary No such oxidase enzyme for GABA is currently available To develop this enzyme Pinnacle will team with an interdisciplinary group of two leading scientists at the University of Kansas Professor Mark Richter is an expert in protein engineering and protein folding and Dr Philip Gao is the Director of the Protein Production Core Facility This team has already cloned expressed purified and characterized an oxidase enzyme wt pUUB Ox with some GABA activity During Phase I we will use this oxidase enzyme as a starting scaffold to evolve a true GABA oxidase enzyme At the end of Phase we will have an oxidase enzyme with a x x improvement in GABA activity relative to wt pUUB Ox and a clear path for Phase II to oxidase activity and stability suitable for the specific measurement of physiologically relevant GABA concentrations This evolved GABA oxidase enzyme will in a single reaction step oxidize GABA to produce hydrogen peroxide as a byproduct The GABA oxidase enzyme can be used as the basis for new monitoring paradigms that would otherwise be impossible By the end of Phase II two commercially available products will be available First a GABA biosensor for real time measurement of physiologically relevant levels of GABA in the brain for preclinical models and second a GABA oxidase enzyme for use in a variety of diagnostic and point of care devices PUBLIC HEALTH RELEVANCE GABA is the major inhibitory neurotransmitter in the brain and plays an important role in disorders ranging from newborn seizures to anxiety Alzheimerandapos s Huntingtonandapos s Parkinsonandapos s diseases and a wide variety of cancers The efficacy of disease models in research is well established for the development of treatments The quality of life and economic costs of these and other illnesses in which GABA plays a role are staggering These disorders disrupt millions of lives and America spends billions of dollars each year in hospital visits nursing home stays and lost productivity