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Lawrence, KS, United States

Patent
Pinnacle Technology, Inc. | Date: 2011-12-01

A biosensor system comprises a capillary substrate, conductive electrode, and a plurality of nanoparticles having an enzyme deposited thereon formed in a cavity at one end of the capillary substrate. The substrate may have an optional reinforcing layer (which may be conductive or non-conductive) and optional insulating layer thereon. A cannula having an optional conductive layer, insulating layer, and reference electrode may also form part of the system.


Grant
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


Grant
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.


Patent
Pinnacle Technology, Inc. | Date: 2011-09-12

The present invention is broadly concerned with the crafting and manufacturability of an implantable enzymatic-based sensor characterized by a small size, optimum geometry, linearity of response over the concentration range of interest, extended shelf-life, selectivity for the analyte in question, and the ability to exclude bioactive interferents. More particularly, it is preferably concerned with a general approach to optimize the performance of the biorecognition elements required to produce biosensors of the type designed to provide, and in conjunction with a suitable signal processing unit, a current which is proportional to the concentration of the analyte of interest. The biosensors described herein may be implanted in vivo, including intra-cerebral, sub-cutaneous, intra-muscular, inter-peritoneal oral, serum, and vascular implantation, the majority of which may act as a surrogate for systemic monitoring and used to monitor analytes of interest in real-time. Multiple biosensors can be joined together to allow for the simultaneous recording of multiple analytes of interest. In addition to the in vivo applications, sensors of the design described herein may also find use in medical monitoring, industrial processes, fermentation, environmental monitoring, and waste water stream monitoring. The present invention offers co-factor enhancement of the biorecognition element, providing access to a range of biorecognition elements heretofore difficult to incorporate into a manufacturing process for the large-scale production of biosensors.


Grant
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

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