The document discusses protein biomarker detection capabilities using nanoparticle probes. It describes a range of protein biomarker concentrations that can be detected, from millimolar to zeptomolar, and examples of disease areas like cancer, Alzheimer's, and cardiovascular disease. It also outlines opportunities for protein biomarker tests in areas like autoimmune disease, pharmacogenomics, respiratory infections, and cardiac biomarkers. The technology uses gold nanoparticle probes attached to capture antibodies to detect proteins through light scattering techniques.

1. Protein
Biomarker
Capabilities

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10-3 – Millimolar
Colorimetric/Enzymatic Chemistry
Blood Sugar (Glucose)
Molecules/Drop
Quadrillions
Nanosphere Technology
Cancer (ovarian, breast and
others), Alzheimer’s, Pulmonary
and Cardiovascular Disease
10-21 – Zeptomolar
10-18 – Attomolar
10-15 – Femtomolar
ELISA & Chemiluminesence
Troponin, CK-MB, BNP, HCG
10-12 – Picomolar
10-9 – Nanomolar
10-6 – Micromolar
Concentration
Trillions
Millions
Billions
Thousands
Tens
<1

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Illustrative Opportunities
Autoimmune
Immune /
Antibodies
Pharmacogenomics
(e.g., 2C19)
Direct
Genomic
Detection
Respiratory
(e.g., Flu,
RSV)
Cardiac
(e.g., cTnI)
High-
sensitivity
Proteins
* Bloodstream infections

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Protein probes use
. Direct antibody attachment
. Biotinilated with free
streptavidin
(published patent app)
. Covalently bound
streptavidin (proprietary
invention to be filed)

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Gold Nanoparticle Probe-Based
– Nucleic Acid Example
A
B
C
D
Detection in the Verigene System relies on the high sensitivity offered by gold nanoparticle probe technology
(A) Target specific capture oligonucleotides are immobilized on a substrate.
(B) Nucleic acid target is presented to the oligonucleotide array substrate under test conditions.
(C) The target is sandwiched by target-specific capture oligonucleotides and DNA-modified gold nanoparticle probes.
(D) Elemental silver is deposited at test site. Light scatter is measured from test sites for detection.

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Sample to result
Simple and easy to use
Random Access
High Clinical Multiplex – 50+

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Sample
Extraction
Amplification
Detection
Extraction
Tray
Amplification Tray Tip Holder
Test
Cartridge
Verigene® SP Consumables

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Fluidics valve
plate
Fluidics
manifold
• Base and
cover
Reagent container
• Cover and
housing
Cartridge
cover
Glass
Substrate
Substrate
holder
RESTRICTED

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Analysis of results (array scanning) occurs via a photon emission
technique where the slide is illuminated internally with light parallel to
the slide surface. Spots where silver-enhanced gold nanoparticle
probes are present emit light. The photons are detected optically and
translated into “signal”.
Light
source

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150 µm
diameter
240 µm grid
RESTRICTED
Target
Potential for additional
targets

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“Serial changes in high-sensitive troponin I predict outcome in patients with decompensated heart failure,” European Journal of Heart Failure (2010) 13, 37–42

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Doxorubicin - Progressive changes observed over time/dose:
1. Lymphopenia/eosinopenia
2. Anemia
3. Nephrotoxicity (glomerular nephropathy)
4. Hyperlipidemia
5. Albumin /globulin
6. ALP and AST changes - likely linked to decreased FC/
bodyweight and mobility, respectively
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Serum cTnI pg/mL
Individual male animals n=67

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Group mean cTnI (pgs/mL) + 1SD
Doxorubicin Dosing Schedule
Number of
animals per
group =6

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19577 1958219581195801957919578
Animal
Number
Serum cTnI pg/mL
Mild atrial
vacuolation
Mild atrial
vacuolation
Mild ventricular
vacuolation
NAD NAD
Minimal focal
necrosis

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ROC curve AUC = 0.92
Evaluation of 131 clinical serum samples (59 non-cancer,
72 cancer)
Plot of ROC Curve

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Test Area 95% CI SE Class = 1
Ave-DNA 0.95 0.93 to 0.98 0.013 have higher values
Ave-protein 0.85 0.81 to 0.89 0.021 have higher values
Com-Ave 0.97 0.95 to 0.99 0.009 have higher values
Results of 310 Patient Samples

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RA assay
• Multiple epitopes for RA test.
• Cirtrulline-, non-cirtrulline compounds, and Ab as
captures
• Performed tests on 1535 patient sera
• Performed comparative assay (Inova 3.1 and RF-ELISA)
• Demonstrated analytical and clinical sensitivity
RA Test chip with 98 captures

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State-of-the-art Competitive Assay
State-of-the-art
Competitive Assay
Rule
out

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• High sensitivity (LOD 0.01 kU/L)
• Ability to use crude allergen
extracts
• Multiplexed microarray
• Rapid assay time (1 1/2 hours)
• Correlated with Phadia’s tests
Microarray with Crude Allergen Extracts Example of an Allergen Test

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