Chemical, Biological, Radiological & Explosive (CBRE) Detection and Protection

1. Chemical, Biological, Radiological & Explosive (CBRE) Detection and Protection Dr. Clifford Lau ODUSD(LABS) 703-696-0371 clifford.lau@osd.mil 27 January 2004

2. DoD Impact Nanotechnology will enable warfighting capabilities * Chem-bio warfare defense Sensors with improved detection sensitivity and selectivity, decontamination * Protective armors for the warrior Strong, light-weight bullet-stopping armors * Reduction in weight of warfighting equipment Miniaturization of sensors, computers, comm devices, and power supplies * High performance platforms and weapons Greater stealth, higher strength light-weight materials and structures * High performance information technology Nanoelectronics for computers, memory, and information systems * Energy and energetic materials Energetic nano-particles for fast release explosives and slow release propellants * Uninhabited vehicles, miniature satellites Miniaturization to reduce payload, increased endurance and range

3. Why nanotechnology and CBRE? • National and Homeland Security • Weapons of Mass Destruction • Chem/bio Warfare Defense • Warfighter and first responder protection • Nanostructures offer unprecedented potential • Sensors with high sensitivity and selectivity • Protection, neutralization, and decontamination

4. OBJECTIVES & DoD IMPACT Develop an experimental and theoretical understanding of the physical and chemical properties of nanoparticle probes functionalized with biomolecules. Engineer Chip-based biodetection platforms. Design and interface a state-of-the-art microfluidic and gel separation system with the chip-based detection platforms. Create handheld biodetection systems for BWA’s, which do not rely on PCR. Massive multiplexing capabilities. Field deployable, PCR-less identification of biowarfare and terrorism agents. Ultrasensitive and Selective Chip-Based Detection of DNAPrincipal Investigator: Chad A. Mirkin,Northwestern University APPROACH • Develop novel detection schemes based on nanoparticle probes to detect specific DNA sequences. • Develop microfluidic systems to isolate cellular DNA from complex biofluidic specimens. • Integrate microfluidic purification, probe/target assembly, and signal transduction features into a single analytical platform. • Investigate the fundamental basis of the selectivity of oligonucleotide-functionalized nanoparticles in chip-based formats using a combined experimental and theoretical approach. • Develop new DNA detection assays based upon metallic and semiconductor quantum dot particles. TECHNICAL ACCOMPLISHMENTS • Developed a novel approach, Biobarcode PCR, for ultrasensitive protein detection. • Developed a Raman labeling technique useful for DNA detection in a random bead array format. • Designed novel copolymer networks and separated proteins from DNA in such networks via microchannel electrophoresis. • Developed a technology for embedding micro magnetic stirrers in Parylene surface micromachined channels. • Developed a technology for making high-density valves and pumps with pressure of membrane displacement  20kPa. • Performed molecular simulations and determined the ion distributions around duplex DNA, and dimers of duplex DNA molecules. • Developed a formally appropriate theory for capacitive charging that describes separate contributions of the DNA transport and the dot charging to the overall conductance.

5. Chemical & biological sensors with improved sensitivity (single molecule) and specificity (no false positives) Explosives & mine detection NRL Nanobiosensors Single protein and nucleic acid molecules (e.g. aptamers and ion channels), single cells, nanoparticles, and nanostructured materials/devices are being characterized and employed for use as sensors of chemical and biological analytes, including use in the stochastic sensing mode that characterizes single molecular binding events. Impact: Sensors to detect and identify unknown analytes.

6. Transparent substrate with optical detection FDB Piezoresistive cantilever FABS Magnetoresistive elements BARC Single Molecule Biosensors Force Discrimination Assay Biosensor Platforms D.R. Baselt, et al., Proc. IEEE85, 672 (1997) G.U. Lee, et al., Anal. Biochem. 287, 261 (2000) M.M. Miller, et al., J. Mag. Mag. Mat.225, 138 (2001)

7. 2 µm 200 µm 4.5 mm Bead Array CounterLloyd Whitman, Naval Research Laboratory Objective: • Develop optics-free DNA chip biosensor with enough sensitivity to eliminate need for PCR amplification Payoff: • Combines state-of-the-art gene chip technology with NRL’s MRAM (magnetoresistive memory) program • Current BARC sensitivity is ~1800 molecules • Current BARC chip has 64-sensing elements for multi-analyte detection Transitions: • Advanced prototype (funded by TSWG) available in FY05 • NRL force discrimination assay/ biosensor technology under CRADA/ license negotiation by several companies 64-sensor BARC chip & next generation instrument • Concept: • Uses DNA-based hybridization assay to detect & identify BW agents • But uses a magnetic bead to label the hybridization reaction • Bound magnetic beads detected with embedded magnetic sensor in the chip • Plan to add immunoassay on same chip J.C. Rife, et al., Sensors & Actuators A107, 209 (2003)

8. Development of Biosensors for Detecting TNT in Seawater Homme W. Hellinga, Duke University Medical Center • Objective: • Redesign the specificity of E. coli periplasmic binding proteins to bind TNT instead of their natural ligands. Approach: Members of the periplasmic binding protein family have been engineered to incorporate fluorescent or electrochemical reporter groups Computational techniques are used to predict the necessary mutations. TNT • Accomplishments: • • Three different receptors were successfully designed to bind TNT. One of these has a 1 nM dissociation constant, sufficient to detect TNT plume edges via UUV. • • More thermostable receptors are being obtained through a combination of rational design and directed evolution. • Methods for immobilization onto surfaces are being refined.

9. Stochastic Chemical Sensing MechanismsHagan Bayley, Texas A&M Cd Zn Co Cd Zn Cd Co The Principle:Single Channel Ion Conductance The Nanomachine:-Hemolysin Channel Genetically Engineered M++ site Analytes + ion flow Transiently bound analyte blocks ion flow Lipid bilayer Open site pA av av analyte-bound site msec Infinitely engineerable (e.g. M++ Site) Analyte Signature = 1/koff Analyte Concentration = 1/kon [analyte] • Issues Under Investigation • Display options (supported bilayers, nanotubular membranes) • Interrogation (microwave, optical) • Multi-valent oligosaccharide receptors • Fluidics (M/NEMS) • Performance • digital, information-rich output • real chemical time, reagentless, self-calibrating • large dynamic range, no signal loss • large analyte universe • M,++ organics, proteins, DNA, (viruses) • Transitions • • Full patent filed • Commercial ventures planned • • DoD Joint S&T Panel for CB Defense award Ternary M++ Mixture

10. MURI: Photocatalytically Active Nanoscale Scavengers and Sensors for CW and Biological Agents Prof. John Yates, University of Pittsburgh GOAL – Develop a multilayer film structure to simultaneously sense and destroy chemical and biological warfare agents. Schematic Multilayer Scavenger and Sensor Device • DELIVERABLES – • Polymer-anchored enzyme and antibody scavengers and sensors for CB agents • Visible-light activated doped-TiO2 nanoparticles with non-photoreactive porous polymer support catalyzing the destruction of both chemical and biological agents • Extremely active CaO and MgO and MgO-Cl2 nanoparticle material for the degradation of CB agents, supported in polymer films. • CHALLENGES – • 1. Integration of both chemical and biological agent sensors and CB catalysts for their destruction into stable multifunctional films or coatings • 2. Efficient photocatalysis in the visible spectrum • 3. Integration of the multifunction films into working devices. • PRIOR WORK – • University of Pittsburgh research on TiO2 photocatalysts and polymer anchored enzymes and sensors. • Kansas State University work on active nanoparticle oxide adsorbents. • Texas A&M University work on antibody-based scavengers.

11. REACTIVE METAL OXIDE NANOPARTICLES FOR SOLDIER PROTECTION Research Objective: Synthesis, characterization, and application of reactive metal oxide nanoparticles for protection against chemical and biological warfare agents and ballistic protection Transitions: ARO program supportsProfessor Ken Klabundeat Kansas State University collaborating with ECBC Next Generation Sorbent Decontamination Program(FY09), Domestic Preparedness Office at SBCCOM, Reactive Protective Skin Cream program at USAMRICD, PM for Nonstockpile Chemical Demilitarization, Ballistic protection programs at Natick Soldier Center. R&D Partners: ARL-ARO/ECBC/NSC/USAMRICD/Universities/Industry New Solutions for Decontamination and Protection of the Soldier in a Chemical and Biological Warfare Environment • Surface area MgO • Commercially Available 30m2 • Reactive Nanoparticles 500m2 • High Surface Area with increased • reactivity Payoff:Highly effective enhanced reactivity for the degradation of chemical and biological agents with reduced materiel burden. Enhanced Ballistic Protection for the soldier.

12. CBRE Grand Challenge Workshop • Workshop held on May 2-3, 2002 in Monterey, CA • In conjunction with AVS meeting • Attended by about 20 participants • Goal was to recommend to NNI a plan of action aimed at realizing the promise of the CBRE grand challenge • CBRE workshop report is available from NNCO

13. CBRE agents • Botulinum Toxin • Diphtheria Toxin • Ricin • Anthrax • Smallpox • Nerve gas (VX, GX, mustard gas, sarin gas, etc.) • Blood agents (hydrogen cyanide, cyanogen chloride, etc.) • TNT • RDX • Plastic explosives • Plutonium • Dirty bombs • Many, many other agents

14. Lethality • Lethal Dosage varies, but can be as low as 0.001 mg/kg body weight • Biological agents are more lethal due to self-replication in the body • Body reaction time can vary from minutes to hours to days to months, depending on the agent • Protection methods also vary

15. Detection • Requirement for nerve agent detection threshold can vary, but can be as low as 0.001 mg/m3 • Required detection time can be as short as less than 10 seconds • Difficult problems • Sensitivity • Sample collection • Liquid or airborne • Selectivity • False alarms • Remote detection

16. Protection • Filtration and Separation • Gas masks, HEPA filters, bullet vests, lead shields etc. • Decontamination and neutralization • Reactive agents (e.g. MgO, Cl, etc.) • RF and plasma techniques • Catalytic nanostructures • Mitigation • After attack • Envionmental issues • Cleansing of filters, sensors, etc.

17. Metrology and Instrumentation Needs • Determination of lethality • Are there other physical properties for detection? • Sensitivity verification • ppm or ppb or ppt is not good enough • Selectivity verification • Different strains of a virus • Protection verification