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Spreeta sensing components

Real-time Monitoring with a Portable Miniaturized Surface Plasmon Resonance System Clement E. Furlong, Research Professor, Departments of Medicine (Div. Medical Genetics) & Genome Sciences University of Washington, Seattle, WA Presented by: Brian Marquardt CPAC/UW. Spreeta sensing components.

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Spreeta sensing components

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  1. Real-time Monitoring with a Portable Miniaturized Surface Plasmon Resonance SystemClement E. Furlong, Research Professor, Departments of Medicine (Div. Medical Genetics) & Genome SciencesUniversity of Washington, Seattle, WAPresented by: Brian Marquardt CPAC/UW

  2. Spreeta sensing components • Spreeta SPR components developed in collaboration with UW with TI • Miniaturized, robust, high performance devices. • Inexpensive in large quantity • Excellent manufacturing capabilities and quality control Each Spreeta chip contains all of the optical components needed for sensitive SPR measurement of biomolecular interactions

  3. The SPIRIT system(Surface Plasmon Instrumentation for the Rapid Identification of Toxins) • Compact, lightweight (lunchbox size, 6 lb.) • High performance • 24 simultaneous measurements • Low power (5W) allows portable operation • Automated Current laboratory prototype

  4. Touchscreen data display Sensor channel Detected levels (bargraph) Selected SPR curve Detected levels (numeric)

  5. Sensor surface chemistry Each Spreeta chip has 3 useable channels Soluble protective coating(dextran/trehalose) allows long-term dry storage at room temperature Control receptors (usually antibodies)Designed NOT to respond to that agent • Target receptors:(usually antibodies)Designed to capture a specific agent or analyte e.g.: • Toxins • Viruses • Spores • Bacteria Gold layer (50 nM) Y Y Y Y YY Y Y Y Y Glass substrate Spreeta sensor chip

  6. Fundamentals of Surface Plasmon Resonance System software Sensorgram

  7. Detection event Analyte SPIRIT performs 24 simultaneous measurements of antibody binding Eight sensor chips Three active spots per sensor Flowcell

  8. Examples of Assays Possible with SPR • Whole microbial cells -(F.tularensis, E. coli, Y. pestis) • Spores -(e.g., anthrax) • Viruses with or without amplification -(e.g. Norwalk, flu) • Proteins by direct detection with or without amplification/verification-(protein toxins, industrial proteins, therapeutics) • Small molecular weight analytes using displacement or competition assays-(e.g., domoic acid, cortisol, insecticides, toxic chemicals, TNT & other small organics)

  9. Detection of Larger Analytes • Microbes • Spores • Viruses • Proteins/Toxic Proteins

  10. Analyte Detection and Signal Amplification Signal Detection

  11. Analyte Detection and Signal Amplification Signal Detection

  12. Analyte Detection and Signal Amplification Signal Detection

  13. Analyte Detection and Signal Amplification Signal Detection

  14. Detection of Microbes Amplification/verification Detection

  15. Virus Detection Amplification

  16. Detection of Staphylococcal Enterotoxin B

  17. Detection of 5 ng/mL (5 ppb; 33pM) BotNT (denatured botulinum toxin)

  18. Direct Detection of Ricin A Chain (64 ppb-320 ppb)

  19. Lower arrows indicate return to no analyte Detection of Cortisol by Competition Assay

  20. Standard Domoic Acid Concentration Curve in Clam Extracts

  21. Other Useful Applications of SPR Sensing Nucleic Acid Analyses Many Other Molecular Interactions

  22. Protein Nucleic Acids as Recognition Elements for DNA/RNA Very stable receptor on chip (Protein Nucleic Acid) Allows detection of target

  23. Binding of a 79 bp DNA Probe to a Complementary PNA 16 mer on the Sensor Surface

  24. Detection of Analytes in Complex Matrices(e.g., saliva, plasma, urine, stool extracts, sea water, fresh water, etc.)

  25. Detection of 1 nM (28 ppb) SEB in seawaterStaphylococcal enterotoxin B

  26. Detection of 500 pM (14 ppb) SEB in urine Amplification Wash(urine) 500 pM SEB From: Naimushin et al., Biosensors and Bioelectronics 17:573

  27. Detection of cortisol in saliva using the compound flow cell

  28. Detection of Theophylline in SalivaUsing the Compound flow Cell

  29. Sequential Detection of 8 Analytes Ovalbumin 10 ng/ml F. tularensis 5 x 103 CFU/ml Norwalk VLPs 5 x 109 particles/ml SEB 5 ng/ml BG Spores 9 x 104 CFU/ml Y. pestis 106 CFU/ml Ricin A chain 20 ng/ml B. anthracis 5 x 106 CFU/ml

  30. SPIRIT Team & Sponsors • Medical Genetics Group: Dr. Clement Furlong Scott Soelberg Dr. Gary Geiss Dr. Rick Stevens Steve Near Matthew Probert Joshua Probert Nathaneal Swanson Dr. Paul Baker • Electrical Engineering Group: Dr. Sinclair Yee Tim Chinowsky Peter Kauffman Jared Tritz Michael Grow Tony Mactutis • Texas Instruments: Jerry Elkind Dwight Bartholomew John Quinn • Sponsors: • DOD • Texas Instruments • Center for Process Analytical Chemistry (CPAC), UW, Seattle • Washington State Sea Grant, NIH/NIEHS

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