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Chemical, Biological, Radiological & Explosive (CBRE)

Detection and Protection

Dr. Clifford LauODUSD(LABS)703-696-0371

clifford.lau@osd.mil

27 January 2004

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Nanotechnology will enable warfighting capabilities

* Chem-bio warfare defenseSensors with improved detection sensitivity and selectivity, decontamination

* Protective armors for the warriorStrong, light-weight bullet-stopping armors

* Reduction in weight of warfighting equipmentMiniaturization of sensors, computers, comm devices, and power supplies

* High performance platforms and weaponsGreater stealth, higher strength light-weight materials and structures

* High performance information technologyNanoelectronics for computers, memory, and information systems

* Energy and energetic materialsEnergetic nano-particles for fast release explosives and slow release propellants

* Uninhabited vehicles, miniature satellitesMiniaturization to reduce payload, increased endurance and range

DoD Impact

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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

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Ultrasensitive and Selective Chip-Based Detection of DNAPrincipal Investigator: Chad A. Mirkin,Northwestern University

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.

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.

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• Chemical & biological sensors with improved sensitivity (single molecule) and specificity (no false positives)

• Explosives & mine detection

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.

NRL Nanobiosensors

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Biosensor Platforms

Piezoresistivecantilever

FABS

Transparent substrate with optical detection

FDB

Magnetoresistiveelements

BARCD.R. Baselt, et al., Proc. IEEE 85, 672 (1997)

G.U. Lee, et al., Anal. Biochem. 287, 261 (2000)

M.M. Miller, et al., J. Mag. Mag. Mat. 225, 138 (2001)

Force Discrimination Assay

Single Molecule Biosensors

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Next Generation BARC(under development)

Next Generation BARC(under development)

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

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

64-sensor BARC chip & next generation instrument

4.5 mm 200 µm

2 µm

J.C. Rife, et al., Sensors & Actuators A 107, 209 (2003)

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Development of Biosensors for Detecting TNT in SeawaterHomme 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:1. Members of the periplasmic binding protein family

have been engineered to incorporate fluorescent or electrochemical reporter groups

2. Computational techniques are used to predict the necessary mutations.

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.

TNT

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Stochastic Chemical Sensing MechanismsHagan Bayley, Texas A&M

Infinitely engineerable (e.g. M++ Site)

The Nanomachine: -Hemolysin Channel

Transiently bound analyte

blocks ion flow

Analytes + ion flow

Lipid bilayer

The Principle: Single Channel Ion Conductance Genetically Engineered M++ site

analyte-bound site

av av

Open site

pAmsec

Analyte Concentration = 1/kon [analyte] Analyte Signature = 1/koff

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)

Cd Zn Co Cd Zn CoCd

Ternary M++ Mixture

Transitions• Full patent filed• Commercial ventures planned• DoD Joint S&T Panel for CB Defense award

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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

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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

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Lethality

• Lethal Dosage varies, but can be as low as 0.001 g/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

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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

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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.

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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