Surface plasmon resonance biosensor for the determination ...
Portable Surface Plasmon Resonance …...Surface Plasmon Resonance • Reduced Fluidics Size •...
Transcript of Portable Surface Plasmon Resonance …...Surface Plasmon Resonance • Reduced Fluidics Size •...
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Joshua Probert, Scott D. Soelberg, Peter Kaufmann
and Clement E. Furlong
Departments of Medicine (Div. Medical Genetics) & Genome Sciences, University of Washington
CPAC Summer Institute,
July 21, 2011
Portable Surface Plasmon Resonance
Instrumentation for Rapid, Versatile
Biodetection
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PROBLEM: CHEMICAL AND BIOLOGICAL HAZARDS A Growing Challenge to Public Safety
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Food supply
Terrorism
Environment
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The Challenge – Detection of Molecular Hazards
Anthrax
Melamine
Shiga Toxin
E. coli
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The Challenge – Fast Detection in the Field
Salmonella
Ricin
E. coli
Paralytic & Amnesiac Toxin
Anthrax
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Angle (pixel number)
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ity
θ
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RIU
x 1
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System Software
Fundamentals of
Surface Plasmon Resonance
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• Reduced Fluidics Size
• Compact, lightweight (lunchbox size, 6 lb.)
• Up to 24 simultaneous measurements for military, food industry, medical and other applications
• Low power (5W) allows portable operation with rechargeable battery
• Semi-Automated, Self Contained Storage and Readout
• Touchscreen Interface For Control and Simple Data Display
• Flow cell interfaces with TEC (±0.01 C)
Current laboratory platform
Portable Multi-channel SPR System
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MBARI AUV
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Old and New Fluidics
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Reduced Fluidics Footprint
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Reduced Electronics Footprint
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Current Interface
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New Interface (Mock-Up)
GO
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Magnetic Nanoparticles
• Magnetic 40 nm
nanoparticles improve sensitivity by concentrating, purifying, and amplifying the signal.
• Effective for detection of low levels of Proteins, Small Molecules, Bacteria/viruses and larger (competition) in complex media
400 nm
probing
distance*
Gold (SPR) Surface
Flow Area
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Magnetic Nanoparticles Amplify the SPR signal
Stepwise Detection of Staphylococcus enterotoxin B
(SEB)
1 Sensor
Ready 2 3
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Time (min)
Rela
tive R
IU
10 ng/ml SEB (13 RIU)
50 ug/ml biotinylated
anti-SEB monoclonal
(24 RIU)
Streptavidin
nanobeads (1075 RIU)
1
3
2
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C NO C
NC
B
NO
C NC
A NO C
NC
Buffer Waste
Sensors
Sample +
Nanobead
Mixture
Magnetic Separation
Column
Pump
1) Load Sample
2) Buffer Wash
(Magnet On)
3) Flow to Sensors
(Magnet Off)
Fluidic Diagram for Automated IMS
Separation and Analyte Detection
U
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Servo Motor Controlled Magnet
Magnet OFF Magnet ON
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ANALYTE ATTACHED TO THE SURFACE
•Small Analytes:
•Estriol, Cortisol, Domoate…
•Same analyte on the surface
Detection of small molecules by SPR
requires an indirect Competition Assay
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NO ANALYTE PRESENT
Time
Re
sponse
Competition Assay
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ANALYTE PRESENT
Time
Re
sponse
Competition Assay
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ELISA vs. SPR
0.00%
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60.00%
80.00%
100.00%
120.00%
0.25 2.5 25 250
SPR ELISA
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Domoate Assay
-20
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120
140
0 2 4 6 8 10 12 14
0ng
1ng
10ng
100ng
1000ng
Bulk
Shift
Binding
Area of
Analysis
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Examples of Assays Performed 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 or with 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)
SPR will detect any ligand in sufficient concentration to which a binding partner (usually antibody) has been obtained, i.e.:
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RIU
Y. pestis
106 CFU/ml
SEB 5
ng/ml
F. tularensis
5 x 103
CFU/ml
B. anthracis
5 x 106
CFU/ml
Norwalk VLPs
5 x 109
particles/ml
Ricin A
chain 20
ng/ml
BG Spores
9 x 104 CFU/ml
Ovalbumin
10 ng/ml
Multiplex Measurement Capability:
Sequential Detection of 8 Analytes
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Ongoing Collaborations
• Dr. Laurie Connell, University of Maine-PNA (peptide nucleic acid) probes for detection of Alexandrium (red tide) mRNA for species identification.
• Dr. Jian Payandeh in Professor William Catterall lab, UW – defining the molecular interactions between sodium channels and their toxin inhibitors.
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MBARI
http://www.mbari.org/AUV
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The UW-Furlong Group
• Professor
Dr. Clement Furlong
• Sensor Team
Scott Soelberg
Peter Kaufmann
Joshua Probert
• Biomarker/PON Team Rebecca Richter Dr. Toby Cole Stephanie Suzuki Dr. Rick Stevens
• Sponsors:
Center for Process Analytical Chemistry (CPAC), UW, Seattle DOD Texas Instruments Washington State Sea Grant NSF/NIEHS NW Center for Human Health and Ocean Studies