Brooks B.D.,Wasatch Microfluidics |
Brooks A.E.,University of Utah |
Brooks A.E.,North Dakota State University
Advanced Drug Delivery Reviews | Year: 2014
With multidrug resistant bacteria on the rise, new antibiotic approaches are required. Although a number of new small molecule antibiotics are currently in the development pipeline with many more in preclinical development, the clinical options and practices for infection control must be expanded. Biologics and non-antibiotic adjuvants offer this opportunity for expansion. Nevertheless, to avoid known mechanisms of resistance, intelligent combination approaches for multiple simultaneous and complimentary therapies must be designed. Combination approaches should extend beyond biologically active molecules to include smart controlled delivery strategies. Infection control must integrate antimicrobial stewardship, new antibiotic molecules, biologics, and delivery strategies into effective combination therapies designed to 1) fight the infection, 2) avoid resistance, and 3) protect the natural microbiome. This review explores these developing strategies in the context of circumventing current mechanisms of resistance. © 2014 Elsevier B.V.
Agency: Department of Health and Human Services | Branch: | Program: STTR | Phase: Phase II | Award Amount: 811.09K | Year: 2011
DESCRIPTION (provided by applicant): The goal of this Phase II STTR project is to develop a real-time label-free biosensor that can analyze 96 samples at a time, compared to the 6 samples possible with current technologies. This platform will initially bedemonstrated with G-protein-coupled receptors (GPCRs) and antibodies. What both applications have in common is the need for higher- throughput sensing, and demonstrating the integrated system for these will illustrate its versatility and potential contributions across the wide spectrum of biosensor applications. For the GPCR demonstration, our work with standard SPR instruments has shown that the choice of detergent(s) is critical for obtaining active solubilized receptor. However, standard low throughput SPR biosensors have two overwhelming drawbacks: (1) The analysis of 96 solubilization conditions requires more than two days and the receptor loses significant activity during this time, which makes it difficult to compare the results obtained at the beginning and end of the analysis and (2) the SPR instrument is limited to testing only one analysis buffer at a time, which means that the success of the entire assay depends on the initial choice of analysis buffer. In Phase I, we developed a 96-channel Continuous Flow Microspotter (CFM) printhead and demonstrated the ability to print GPCRs onto a sensor surface directly from crude media using up to 96 different analysis buffers. The GPCRs were also kept wetted and active throughout the printing process by theCFM's enclosed microchannel printing network. In the final experiment, we solubilized the GPCR CCR5 from whole cells using 192 different detergent conditions and spotted them onto an SPR sensor surface using our CFM printhead. We then tested the activity of the receptor and used the ligand binding results to determine that a certain combination of detergents best enhanced receptor activity. To run this analysis with a standard Biacore technology (e.g. T100) would have required four days of instrument time.In comparison, we were able to perform the analysis in less than 2 hours. In Phase II, we propose to integrate the 96-channel CFM from Phase I with a commercial SPR imager to enable automated interaction analysis in a highly parallel format. The followingspecific aims detail the combination of Wasatch's microfluidic technologies with the commercial IBIS SPR imager to produce a high-throughput label-free biosensor. 1. Refine the 96-channel CFM printhead design from Phase I to enable optimal performance whenmounted on the IBIS SPR imager. 2. Mount the 96-channel CFM onto the IBIS SPR imager and optimize the fluidic parameters that affect platform sensitivity and uniformity. 3. Automate the CFM and SPR components within one seamless instrument: Automation ofthe flow cell positioning, sealing, fluid handling, valving, in-line degassing and temperature control. Integration of the CFM and SPR imager control software and data collection/analysis software. 4. Demonstrate use of the automated system with GPCRs andantibodies. PUBLIC HEALTH RELEVANCE: Antibody analyses are one of the most common applications of biosensor technology and are typically straightforward. GPCRs are the hottest and most challenging system that biosensor users are tackling. Up to halfof the drugs on the market today modulate some form of GPCR activity, and it is estimated that 25-50% of the total drug targets are in the GPCR families. GPCRs are the most studied of the major drug target classes, yet they are challenging to work with because they are normally membrane associated, present in low abundance, and unstable. By enabling the high throughput study of GPCRs, there is enormous potential for speeding drug development, treatments, and the associated health of patients with hundredsof different diseases.
Brooks B.D.,Wasatch Microfluidics
Current Drug Discovery Technologies | Year: 2014
The pharmaceutical industry is experiencing comeback sales growth due in large part to the industry's R&D efforts that center on biologics drug development. To facilitate that effort, tools are being developed for more effective biologic drug development. At the forefront of this effort is epitope characterization, in particular epitope binning, primarily due to the role an epitope plays in drug function. Here we detail the financial advantages of epitope binning including (1) increased R&D productivity due to increased work in process, (2) reduced number of "dead-end"candidates, and (3) increasedability to reengineer antibodies based on the epitope. With the arrival of high throughput biosensors, this manuscript serves as a call to push epitope binning earlier in the biological drug discovery process. © 2014 Bentham Science Publishers.
Wasatch Microfluidics and Rinat Neuroscience | Date: 2014-12-02
A system and method for sensing and analyzing antibody blocking interactions is described. A biosensor can be used to identify interactions between antibodies to generate interaction profiles for the antibodies. A processor can be used to assign the antibodies to one or more bins, with the antibodies sharing a common interaction profile assigned to a common bin, and each antibody only being assigned to one bin. The antibodies can be represented by displaying nodes grouped together for antibodies in a common bin. Connections between the nodes can be displayed, representing interactions between the antibodies.
Wasatch Microfluidics | Date: 2015-03-24
A system for depositing substances onto a deposition surface can comprise a first contact spotter comprising multiple spotting orifices fed by multiple fluid inlet conduits such that the first contact spotter is capable of depositing multiple spots of different substances onto the deposition surface simultaneously, and a second contact spotter comprising a second spotting orifice fed by a second fluid inlet conduit. The system can also include a positioning device adapted to alternatively position and seal the first contact spotter and second contact spotter on the deposition surface at an overlapping location.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 225.00K | Year: 2014
Project Summary We propose an instrument system for automated, multiplexed cell- and tissue-based experiments (i.e. tissue microarrays) called the Microfluidic Flow Cell Array (MFCA). The MFCA consists of a microfluidic flow cell array integrated with an inverted fluorescent microscope, allowing the observation of 48 flow chambers simultaneously or an individual chamber at higher magnification. The proposed instrument will be used to novel cell- and tissue-based assays in a highly parallel manner that are otherwise difficult to perform. Our goal is to build a platform for producing a tissue microarray that maintains many of the characteristics found in cells in vivo and allows for the investigation of cell-surface, cell-cell, and especially tumor-stroma interactions, cell toxicity studies, and tissue toxicity studies in a multiplexed fashion. As a Phase I application, this proposal's focus is ovarian cancer tissue slice toxicity testing to determine the optimal chemotherapeutic agents for a specific patient
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase II | Award Amount: 751.91K | Year: 2016
Abstract Vaccines are fundamental in preventing debilitating illnesses and saving millions of lives each year But despite these vaccine oriented victories infectious diseases are still the second leading cause of death as well as the leading cause of disability adjusted life years worldwide About six million deaths were attributed to AIDS malaria and tuberculosis in regions where no vaccine is currently available Other serious infectious diseases without vaccines include many sexually transmitted diseases as well as parasitic respiratory and enteric diseases that afflict millions of people annually More concerning is that the world must now cope with new and re emerging diseases and increasing antimicrobial resistance Governments have invested heavily in vaccine development over the past decades however many diseases remain challenging primarily due to the evolved mechanisms of evasion that pathogens develop Iterative approaches to vaccine development are necessary to overcome these mechanisms of evasion Iterative approaches require combination approaches discovery of novel antigens adjuvant and vectors in the preclinical stage with computational analyses of clinical data to accelerate vaccine design To facilitate these efforts it is imperative to develop tools to get more and better information to the vaccine engineers faster so that the next generation of vaccine can be developed Wasatch proposes in this Direct to Phase II SBIR project to develop a reengineered Surface Plasmon Resonance SPR platform that will pursue next generation features for both vaccine research and increased throughput to get kinetics and epitope information to vaccine engineers early in the vaccine development process The project directly addresses the focus areas of special SBIR solicitation Direct Phase II SBIR Grants to Support Biomedical Technology Development This new SPR system would allow direct from serum information to be gathered to improve the quality of the data and to greatly decrease the time required to gather it We propose to significantly advance the current Wasatch Microfluidics SPR platform which provide a solid foundation via redesigned optics for improved sensitivity individually valved channels to print optimal amounts of serum and custom next generation software development effort to process the large amounts of data generated In high throughput screening mode our channel integrated SPR platform will be capable of collecting data from andgt samples in less than hours vs the current samples per day Wasatch Microfluidics Gary Cohen s group at the University of Pennsylvania and David Myszka of Biosensor Tools will pursue two Specific Aims Engineer a channel SPRi array instrument to analyze simultaneous multiplexed antigen vaccine serum samples with advances microfluidics imaging and software and Perform a vaccine characterization study by capturing total IgG out of guinea pig sera vaccinated and or challenged with herpes virus Project Narrative Vaccine research has received increasing interest due to the threat of emerging diseases unique applications of vaccines such as cancer and the effectiveness of past vaccines however new vaccine development will require innovative new approaches built on immunogenicity modeling We propose to develop high throughput biosensors with improved throughput sensitivity and data processing for immunogenicity modeling to give researchers the opportunity to develop to create vaccines against communicable and highly adaptive viruses as well as unique diseases such as cancer Vaccines research has saved more lives than any other medical advancement and this technology could provide advances to improve the quality of life and to save the lives of countless more individuals
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 329.75K | Year: 2014
DESCRIPTION (provided by applicant): Real-time label-free technologies such as surface plasmon resonance biosensors provide high-resolution information about the kinetics, affinity, stoichiometry, activity, and specificity, of two (or more) binding partners. While the application f biosensors is well established, current instrumentation has limited sampling throughput. Screens of even a fairly small chemical library (e.g., 3000-5000 compounds) require days to weeks to complete using traditional label-free instruments. Because of the quality of the data generated by SPR, drug discovery scientists are clamoring to use enhanced biosensors as a screening tool for small molecule applications; however, lack of throughput hinders their ability to move in this direction. We propose to develop a biosensor platform that has increased throughput yet maintains the data quality and ease of use to which researchers are accustomed. We will couple our novel Continuous Flow Microspotter (CFM) with an enhanced-sensitivity SPR
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 327.66K | Year: 2012
DESCRIPTION (provided by applicant): The goal of this Phase I SBIR project is to develop a device for automated array printing of lipids and membrane proteins onto submerged microtiter plate surfaces in a way that maintains their activity and function. Ourproprietary 3D microfluidic printing technology is uniquely capable of sealing and printing arrays of biomolecules onto submerged surfaces in an automated fashion. By printing onto submerged surfaces, we will produce lipid arrays that maintain many of the characteristics found in cellular membranes and enable the investigation of protein-ligand and protein-membrane interactions in a multiplexed fashion. Such a tool would enable novel lipid-based research and diagnostic assays in a multitude of areas including cancer, diabetes, inflammation, infections, and cardiovascular disease. Wasatch Microfluidics has previously developed a flow-based microfluidic printing technology, the Continuous Flow Microspotter (CFM), which uses 3D channel networks to print biomolecules onto flat surfaces by flowing them back and forth over discrete spot locations. We propose to take the next step by developing a CFM printhead that can print within the bottom of a 96 well microtiter plate, adapting the CFM printer hardware to accept microtiter plates, and developing new hardware, software and techniques for automated printing on submerged surfaces. The following specific aims have been identified to prove the feasibility of using Wasatch's CFM flow printing technology for submergedprinting of lipids and GPCRs within 96 well microtiter plates. 1. Design a CFM printhead that can print within the well of a standard 96 well microtiter plate. 2. Adapt the CFM hardware and software to enable automated submerged printing on the bottom ofmicrotiter wells. 3. Multiplex lipid array printing and analysis in partnership with Echelon Biosciences and David Myszka. PUBLIC HEALTH RELEVANCE: The combination of fluid lipid bilayers and membrane proteins constitute the major structural components of biological cell membranes. All major human diseases including metabolic, autoimmune, vascular, neurological, and cancer have essential pathologic mechanisms involving dysfunctional cell and/or internal membranes. By enabling multiplexed analysis of lipids and membrane proteins using currently available microtiter plate based technologies, there is a tremendous opportunity to study these systems in their native states and thereby improve our knowledge of receptor structures and functions.
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase II | Award Amount: 1.02M | Year: 2015
DESCRIPTION provided by applicant Real time label free technologies such as surface plasmon resonance biosensors provide high resolution information about the kinetics affinity stoichiometry activity and specificity of two or more binding partners While the application f biosensors is well established current instrumentation has limited sampling throughput Screens of even a fairly small chemical library e g compounds require days to weeks to complete using traditional label free instruments Because of the quality of the data generated by SPR drug discovery scientists are clamoring to use enhanced biosensors as a screening tool for small molecule applications however lack of throughput hinders their ability to move in this direction We propose to develop a biosensor platform that has increased throughput yet maintains the data quality and ease of use to which researchers are accustomed We will couple our novel Continuous Flow Microspotter CFM with an enhanced sensitivity SPR biosensor from BiOptix to enable label free screening and kinetic analsyis of small molecules and biologics In high throughput screening mode our channel integrated CFM E biosensor platform will be capable of collecting data for andgt samples in less than hours a sampling rate x faster than fastest small molecule capable label free biosensor the Biacore PUBLIC HEALTH RELEVANCE Label free real time biosensors enable the measurement of the kinetics of biomolecule binding Getting this information earlier in the drug discovery process reduces false positives identifies candidates that may have been missed improves subject matter for patent filings and increases the probability of the selected candidateandapos s eventual success Traditional label free biosensors have limited sample throughput which has restricted their use to secondary roles By combining our highly parallel microfluidic sample delivery technology with a small molecule sensitive biosensor we will enable the high throughput label free analysis of small molecules and biologics which offers significant potential for increasing the efficiency of early stage drug discovery