Transcript Biomemetics - World Technology Evaluation Center (WTEC)

Chemical, Biological,
Radiological & Explosive (CBRE)
Detection and Protection
Dr. Clifford Lau
ODUSD(LABS)
703-696-0371
[email protected]
27 January 2004
7/7/2015
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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
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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 DNA
Principal Investigator: Chad A. Mirkin,Northwestern University
OBJECTIVES & DoD IMPACT
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APPROACH
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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 chipbased formats using a combined experimental and
theoretical approach.
Develop new DNA detection assays based upon metallic
and semiconductor quantum dot particles.
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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.
TECHNICAL ACCOMPLISHMENTS
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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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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.
• Chemical & biological sensors with improved sensitivity
(single molecule) and specificity (no false positives)
• Explosives & mine detection
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Single Molecule Biosensors
Force Discrimination Assay
Biosensor Platforms
Piezoresistive
cantilever
FABS
D.R. Baselt, et al., Proc.
IEEE 85, 672 (1997)
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Transparent substrate Magnetoresistive
elements
with optical detection
BARC
FDB
G.U. Lee, et al., Anal.
Biochem. 287, 261 (2000)
M.M. Miller, et al., J. Mag. Mag.
Mat. 225, 138 (2001)
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Bead Array Counter
Lloyd Whitman, Naval Research Laboratory
2 µm
4.5 mm
200 µm
64-sensor BARC chip & next
generation instrument
Next Generation BARC
(under development)
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
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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
J.C. Rife, et al., Sensors & Actuators A 107, 209 (2003)
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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:
1.
2.
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.
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Stochastic Chemical Sensing Mechanisms
Hagan Bayley, Texas A&M
The Nanomachine: -Hemolysin Channel
The Principle: Single Channel Ion Conductance
Genetically
Engineered M++
site
Analytes + ion flow
Transiently
bound analyte
blocks ion flow
Lipid
bilayer
Open site
pA
Infinitely engineerable (e.g.
M++
av
msec
av
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)
Transitions
• Full patent filed
• Commercial ventures planned
• DoD Joint S&T Panel for CB Defense award
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analytebound site
Performance
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digital, information-rich output
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real chemical time, reagentless, self-calibrating
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large dynamic range, no signal loss
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large analyte universe
M,++ organics, proteins, DNA, (viruses)
Cd
Zn
Co
Cd
Zn
Ternary M++ Mixture
Cd
Co
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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 mg/kg body weight
• Biological agents are more lethal due to selfreplication 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
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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
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