Technologies to Enable Autonomous Detection for BioWatch Ensuring Timely and Accurate Information for Public Health Officials: Workshop Summary (2014) / Chapter Skim
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Appendix H: White Paper 3: State of the Art for Autonomous Detection Systems Using Immunoassays and Protein Signatures
Appendix H: White Paper 3: State of the Art for Autonomous Detection Systems Using Immunoassays and Protein Signatures
Pages 173-196
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GenArraytion, Inc. A white paper prepared for the June 25–26, 2013, workshop on Strategies for Cost-Effective and Flexible Biodetection Systems That Ensure Timely and Accurate Information for Public Health Officials, hosted by the Institute of Medicine's Board on Health Sciences Policy and the National Research Council's Board on Life Sciences.
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rapidly process and accurately analyze aerosol samples with a high level of confidence; (2) automate and integrate the major system functions into the detector, including aerosol sample collection, preparation, analysis, and analytical results reporting; (3)
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The standard IgG immunoglobin is a Y-shaped molecule with the short arms representing identical binding elements and a base that can be anchored to a variety of surfaces. The binding region recognizes a small region, generally three to five amino acids in size, which is unique from the host that generated the antibody as part of an immune response.
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, providing many more independent binding opportunities and dramatically increasing the sensitivity of the system. In this way antibody detection and identification can compete with sensitivities achieved by nucleic-acid amplification technologies.
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Nonspecific binding is caused by a label, or reporter molecule, being localized with the capture antibody, and it creates noise in the system. It is generally accepted that a specific signal must be at least three times the intensity of the background to be a specific binding event.
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The types of reporter systems can be divided into three broad categories. The first is optical reflectance, such as the colloidal label on secondary antibodies utilized in most lateral flow devices.
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Figure H-3 illustrates the detection mechanism for the MagPix system, which uses LEDs for bead optical interrogation and immobilizes the beads with a magnet to allow for increased image capture time, thereby improving sensitivity. Figure H-4 shows the Magpix system, which has a relatively small footprint and has user-friendly interfaces.
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Sample volume uptake 20–200 ml Probe piercing Yes Auto adjust-probe height Yes Daily startup ≤15 minutes Sensitivity Approximately 106 copies of DNA or single-digit picogram levels of protein, 102–103 copies of bacteria. Dynamic range 3.5 logs Strengths The open architecture allows for sig nificant flexibility for generation of additional tests in any laboratory.
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The CANARY® technology shown in Figure H-5 differs significantly from the standard immobilized antibody. This is the only technology discussed in this paper that exploits the natural mechanism of antibody response when a specific antigen is detected.
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of target organisms with a time to result of 3 minutes, including sample processing (see Figure H-6)
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APPENDIX H 183 TABLE H-3 Selected Subset of Available B-Cell Lines with Analytical Sensitivities Antigenic Assay Agent Test Panel Target Sensitivity Bacillus Targets Positive against live 10 cfu/sample anthracis anthrax spore Sterne strain, (Sterne) (spores)
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(RI) has been involved in the BioWatch program for a number of years.
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System operation may also be remotely controlled in real time with virtually every fluid or air handling function being capable of remote manipulation. The bioidentification portion of the instrument can also analyze fluids from other sample collection systems or from liquid samples loaded into the instrument through a manual sample port.
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For more sophisticated users, bundled Windowsbased software allows the development of customized sample collection and detection protocols. The system is designed to be operated over a very wide temperature range, from −32°C to 60°C.
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o Quantitative information is saved in on-board flash memory and may be automatically transmitted to a remote computer if desired. Measurement interval o Aerosol trigger: 1 minute updates o Sample collection and agent identification initiated on alarm from aerosol trigger o Sample collection: 5 minutes minimum recommended for first sample after alarm o Agent identification: 30 minutes assay time for first sample after first collection o Sample collection: user definable after initial collection o Agent identification: 17 minutes for each additional sample Cost o Initial purchase: $125,000 to $150,000 in quantities <10 de pending on exact configuration.
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Battelle REBS Biosensor Technology Battelle has patented an integrated collection and identification spectroscopic technology, the Resource Effective Bio-Identification System (REBS) , which provides a portable platform that is a lightweight, lowcost, networked, battery-operable system with near-real-time identification of environmental aerosols, surface contamination, and waterborne biological warfare agents (BWAs)
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These tests included four BWAs, including gramnegative and gram-positive bacteria, virus, and protein simulants. Battelle has carried out extensive testing to establish the probability of correct identification, first based on Raman spectroscopy of single particles and, second, on the consensus of several identified particles FIGURE H-7 Battelle's Resource Effective Bio-Identification System (REBS)
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REBS requires electrical power from at least one source. Potential power sources include military-style batteries, 120- to 240-VAC (volts alternating current)
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Remote control and monitoring are achieved using a wired or wireless network communication protocol and remote computer graphical user interface. Expandable Threat Identification REBS detects and identifies at the genus, species, and strain levels based on the Raman signatures generated by a laser field interacting with the molecules and structural conformations of biological materials that comprise BWAs.
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Antibody-based assays immobilize the capture antibody in a black plastic well, and then standard sandwich assays are performed with a proprietary reporter light-generating reaction. Electrochemiluminescence detection uses labels that emit light when electrochemically stimulated (see Figure H-8)
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SOURCE: Company website. The system offers multiplexed detection technology by spatially separated spots on the bottom of a 24-, 96-, or 384-well format microtiter plate (see Figure H-9)
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The beads occupied with target are then incubated with a secondary reporter antibody, which is covalently linked to an enzyme. The conjugated enzyme bead complex is rinsed in a solution containing an optically inert substrate.
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(c) Fluorophore concentration in the small sample volume of wells containing the target analyte rapidly reach detectable limits using conventional fluorescence imaging and can be digitally counted.
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196 TECHNOLOGIES TO ENABLE AUTONOMOUS DETECTION FOR BIOWATCH FIGURE H-11 Quanterix Simoa HD-1 analyzer. SUMMARY All of the systems presented have the capability to meet the majority of the autonomous detection system requirements.
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