Download Single Molecule Array (Simoa) technology for ultrasensitive

Single Molecule Array (Simoa) technology for ultrasensitive,
multiplexed detection of proteins and nucleic acids
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Corporate Overview
• Founded 2007 by David Walt and Nicholas Naclerio
• Exclusive licensee of patent portfolio developed by David Walt at Tufts University
• ̴ 50 employees, 15 Ph.D’s
• Expanded to state-of-the-art 20,000 sq. ft. facility April 2012 in Lexington, MA
• Venture-backed by leading life science investors and strategic partners
• Cumulative $47M Raised in Series A, B, C
Board of Directors
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Martin Madaus, Ph.D. (Executive Chairman)
John Connolly (Bain Capital Ventures)
David Walt, Ph.D. (Tufts University, Illumina)
Keith Crandell (ARCH Venture Partners)
Doug Cole, M.D. (Flagship Ventures)
Simon Davidson (In-Q-Tel Ventures)
Dennis Sandstedt (bioMérieux)
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Serum
Concentration
10−3
1x
M to
-12
1 x 10 M
Total Number of
Serum Proteins
All Protein Blood Tests are Based on the
Currently Measurable 250 Proteins
Clinical Chemistry
130 FDA Approved Tests1
2501
ELISA
Quanterix
Opportunity
How many proteins
Are below 1 x 10-12 M?
Potential is Large:
Human Proteome = 25,000 genes
Secreted Proteins >2,500
1Anderson
2Klee
and Anderson, Molecular & Cellular Proteomics 2002: 1.11 845-867
et al., Nucleic Acids Research 2004: 32, 1414-1421
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Single Molecule Arrays (Simoa) Provide a Digital Advantage
0 pM
10 pM
• Reaction volume = 100 ×10−6 L
• Diffusion = dilution = low sensitivity
• Millions of molecules needed to reach detection limit
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Reaction volume = 50 ×10−15 L (2 billion times smaller)
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Diffusion defeated = single molecule resolution = ultimate sensitivity
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One molecule needed to reach detection limit
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Counting Single Protein Molecules in Blood using Simoa
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Simoa Enables Single Molecule Diagnostics
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Simoa Uses Digital Counting and Intensity to Determine Average
Number of Enzymes per Bead (“AEB”)
AEB ~ 0.1
AEB ~ 0.6 AEB ~ 3.0
Rissin et al., Anal.Chem. 2011, 83, 2279-2285
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10
10
AEB of IL-6 beads
AEB of TNF-α
α beads
Multiplexed Single Molecule Immunoassays
1
0.1
0.01
0
0.1
1
10
1
0.1
0.01
100
0
0.1
[TNF-α
α ] (pg/mL)
AEB of IL-1β
β beads
1
0.1
0.01
0.1
1
[IL-1α
α ] (pg/mL)
10
10
100
100
10
100
10
1
0.1
0.01
0
0.1
1
[IL-1β
β ] (pg/mL)
20
[cytokine] (pg/mL)
AEB of IL-1α
α beads
10
0
1
[IL-6] (pg/mL)
15
10
5
0
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Digital ELISA Detects Proteins in the fg/mL Range
Protein
SiMoA LOD
(pg/mL)
Gold Standard
Assay LOD
(pg/mL)
Vendor
Sensitivity
Fold
Increase
IL-1β
TNF-α
GM-CSF
p24
IL-6
PSA
Aβ42
tau
IL-1α
GLP-1
p-tau (181 + 231)
IL-5
troponin
p-tau-231
p-tau-181
0.001
0.003
0.003
0.005
0.005
0.006
0.02
0.02
0.024
0.033
0.04
0.049
0.05
0.05
0.1
0.057
0.106
0.26
15
0.039
10
50
60
1
11
NA
0.115
3
NA
60
R&D Systems
R&D Systems
R&D Systems
OCD
R&D Systems
Roche/Siemens
Innotest
Innotest
R&D Systems
Luminex
NA
R&D Systems
Abbott Architect (HS)
NA
Innotest
57
35
87
3000
8
1667
2500
3000
42
333
ND
2
60
ND
600
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Simoa Addresses Many Opportunities That Exist for
High Sensitivity Clinical and Companion Diagnostics
Neurology
Enables blood tests for
neurological disease
Cardiac
Inflammation
First to quantify effect of antiTNF-α
α drugs on target protein
Infectious Disease
World’s most
sensitive troponin test
First IA as sensitive to
virus as PCR
Signal Transduction
HomeBrew
Oncology
Shorten detection of recurrence
(years to months)
Metabolism
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Application Demonstration: Infectious Disease
HIV (p24 capsid protein)
• Achieved 2,000-fold greater sensitivity than benchmark commercial
immunoassays (LoD = 5 fg/mL), which is equivalent to about 50 copies of
virus per mL. First immunoassay that has shown equivalent sensitivity as
nucleic acid testing (NAT).
• Demonstrated full analytical performance (blinded positive and negative sera,
blinded seroconversion panels, spiked HIV virus etc.)
• Demonstrated SiMoA is as sensitive as PCR for detecting HIV virus in early
infection (seroconversion panels)
• Clinical value: HIV screening for acute
infection (prior to seroconversion) and
monitoring therapy. Blood screening
for infection.
• Commercial leverage: Simplicity, cost
and throughput vs. NAT.
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Detection of DNA using Simoa
[bacteria] in blood (fM)
0
0.067
0.67
6.7
1,000
10,000
100,000
0.4
Streptavidin-βgalactosidase
Substrate
Target sequence
Product
0.3
AEB
Biotinylated detection
sequences
0.2
0.1
0.0
Capture DNA sequence
0
No. bacteria per 25 µ L blood sample
Magnetic
Particle
[bacteria] in river water (fM)
0.033
0.33
1,000
10,000
3.3
33.3
0.4
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Detected S. aureus spiked into whole blood and
Charles River water
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Same sensitivity as PCR (~70 aM)
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Greatly reduced risk of cross-contamination of
samples vs. PCR because direct detection of DNA
AEB
0.3
0.2
0.1
0.0
100,000
1,000,000
No. bacteria per 50 µ L river water sample
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Simoa HD-1 Analyzer
• Unprecedented ultrasensitivity
• Full automation
• Precision
• Wide dynamic range
• Easy & cost-effective implementation
• Multiplexing capability with small sample
volumes
• Homebrew capabilities
• >500 samples per shift
• Available October 2013 for RUO
• IVD clinical studies begin 2014
Sensitivity Means Knowing
Simoa is a high definition diagnostics system
that gives you the power to see more clearly,
know with certainty, and reach new heights
in lab productivity.
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Simoa Disc
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Low-cost consumable
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24 arrays/disc (4 discs = 96 well
plate)
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216,000 femtoliter wells per array
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Manufactured using state-of-the
art Blue-ray™ technology by Sony
DADC
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High volume production easily
scaleable
216K wells
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Quanterix Instrument Workflow
Step 1. Load Consumables
• Kits (singleplex or multiplex)
• Cuvettes and disposable tips
• Simoa discs
Step 2. Menu Setup
• Select assay
• Calibrators and controls
Step 3. Load Samples
• 96 well plates or tubes
Step 4. Run Samples
• Simoa HD-1 Analyzer
performs all mixing, washing,
incubation, and sample readout, with first result in 30
minutes and subsequent
samples every 45 seconds
thereafter.
Ultra-sensitive with
fully automated
simplicity
Multiplexed analysis
and small sample
volumes
Open platform
(Homebrew for RUO)
High volume, low cost
production of 24array discs
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Simoa HD-1 Roadmap
2011
Instrument
Development
2011-13
Menu Dev.
2013
RUO
Launch
2015
Increased plex
DNA, RNA
2016
IVD
SiMoA
Research Use
Only
Companion Dx
In vitro diagnostics
Non-regulated clinical studies
Biomarker validation
Pharma, Biotech, Core Labs, CROs
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Exclusive
Partner
Quanterix Value Map in Biomedical Industry
Clinical
IVD
RUO
Pharma &
BioPharma Clinical
CRO
Novel Markers
Differentiated
Value
Quanterix
Revenue
Streams
Sensitivity – Efficiency – Precision – Multiplexing - Value
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Simoa HD-1 Analzyer
Assay Kits
Homebrew
Service
Boutique Testing & Custom Assay Dev.
Blood
Screening
Quanterix or Co-Exclusive
Academic
Research
LDTs
cDx
POC
Food
Safety,
Biothreat,
etc.
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Thank You
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