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Single Molecule Array (Simoa™) for Ultra High Sensitive Detection of Biomarkers

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Who We Are

Founded in 2007, Quanterix is a leader in high definition diagnostics, delivering ultrasensitive single molecule measurements for the benefit of human health.

• Leverages proprietary single molecule array (Simoa™) technology • Offers 1000x sensitivity improvement over current standard • Proven applications in oncology, neurology, infectious disease, and beyond

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Corporate Overview

• Founded 2007 by David Walt (founder of Illumina) 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

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Quanterix is supported by and working with leading organizations around the world

Investors Partners

Funding Collaborations

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Counting Single Protein Molecules in Blood using Simoa

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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

• Reaction volume = 50 ×10−15 L (2 billion times smaller), 50 femtoliters

• Diffusion defeated = single molecule resolution = ultimate sensitivity

• One molecule needed to reach detection limit

Simoa (Digital)

0 aM 3.5 aM 350 aM 3.5 fM

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Step 1. Load Consumables

• Kits • Cuvettes and disposable tips • Simoa discs

Step 3. Load Samples

• 96 well plates or tubes

Step 2. Menu Setup

• Select assay • Calibrators and controls

Step 4. Run Samples

Step 5. Data analysis

• First result in 75 min

Ultra-sensitive with fully automated simplicity

Small sample volumes

Open platform (Homebrew for RUO)

68 test per hour, >500 data points per day

Quanterix Instrument Workflow

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Simoa Disc

• Low-cost consumable

• 24 arrays/disc (4 discs = 96 well plate)

• 216,000 femtoliter wells per array

• Manufactured using state-of-the art Blueray™ technology by Sony DADC

• High volume production easily scalable 216K wells

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Quanterix Instrument and Assay Features Metric Quanterix

Throughput • 68 Tests/hour • >500 data points per day

Workflow • Batch (plates or tubes)

Total Assay Time • <2.5 hours per 96-well plate

Hands on Time • Startup time <20 minutes • Walkaway time TBD

Automation • Full (sample in to data out)

Sample input • 96-well plates and tubes

Sample volume • 10 ml -100 ml

Instrument Footprint • 135x60x160 cm (freestanding)

Content • 30 Kits at launch • Homebrew (build your own)

Assay Flexibility • 1, 2 or 3-step assays, incubation time, number of wash steps

Metric Quanterix

Sensitivity • 200 - 1,000 fold better than ELISA

Dynamic range • >4 logs

Precision • <10%

Instrument

Assay Performance

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Digital ELISA detects proteins in the fg/mL range

Protein SiMoA LOD

(pg/mL)

Gold Standard Assay LOD

(pg/mL)

Sensitivity Fold

Increase

IL-1b 0.001 0.057 R&D Systems 57

TNF-a 0.003 0.106 R&D Systems 35

GM-CSF 0.003 0.26 R&D Systems 87 p24 0.005 5 OCD 1000

IL-6 0.005 0.039 R&D Systems 8 PSA 0.006 10 Roche/Siemens 1667

Ab42 0.02 50 Innotest 2500

tau 0.02 60 Innotest 3000

IL-1a 0.024 1 R&D Systems 42

GLP-1 0.033 11 Luminex 333 p-tau (181 + 231) 0.04 NA NA ND

IL-5 0.049 0.115 R&D Systems 2 Troponin I 0.05 3 Abbott Architect (HS) 60 p-tau-231 0.05 NA NA ND

p-tau-181 0.1 60 Innotest 600

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Simoa Addresses Many Opportunities That Exist for High Sensitivity Clinical Diagnostics

Infectious Disease

Inflammatory Disease Oncology

1 mm3 Tumor

Blood Brain

Endothelial Cells

Neurodegenerative Disease

Prenatal Screening

Cardiovascular Disease

Abnormal/ Rare Protein

Simoa allows blood tests for

neurological disorders

Simoa is world’s most sensitive

troponin test

Simoa is first Immunoassay to

be as sensitive to virus as PCR

Simoa first technology to

quantify effect of anti-TNF-a

drugs on target protein

Simoa shortened time for

detection of cancer recurrence

from years to months

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Inflammation

Crohn’s Disease (TNF-a, IL-6) (Mayo Clinic)

• Study Objectives: Measure cytokine levels in normal individuals and patients with clinically active Crohn’s disease (CDAI>220) undergoing anti-TNFa therapy (e.g. Remicade, Humira, Enbrel)

• Completed retrospective clinical study on 17 patients.

• Observed significant elevation of IL-6 in plasma from patients compared to controls. After therapy, the mean reduction in the concentrations of TNF-a and IL-6 were 46% and 58%, respectively.

• Clinical Utility: Changes in cytokine concentrations after therapy indicate that measurement using digital ELISA could be used for monitoring therapeutic efficacy

Song et al., J. Immunol. Methods, 2011, 372, 177-186 0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

#1 #3 #5 #11 #13 #17 #19 #25 #27

Case Number

Before treatment with anti-TNF-a

After treatment with anti-TNF-a

TN

F-a

(pg

/mL

)

xMAP LOD

Crohn's Disease Healthy

0.01

0.1

1

10

100

[IL

-6]

(pg

/mL

)

xMAP LOD

Simoa LOD

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Neurology

Brain hypoxia (Ab42, tau) (Univ. of Gothenburg)

• Achieved 2500-3000-fold greater sensitivity than benchmark commercial assays (LOD 10-20 fg/mL). Only test capable of precise measurement of these markers in blood (normally tested in CSF).

• Completed retrospective clinical study on 25 cardiac arrest patients. Objective: look for changes in serum levels of these markers as potential early indicators of long term brain damage.

• Observed, for the first time, Ab42 elevation in live humans associated with brain ischemia. Suspected significance in Alzheimer's disease pathogenesis. Strong correlation with 6 month cognitive outcome. Tau data even stronger.

• Clinical Utility: early prognostic indicators of long term brain damage.

Zetterberg et al., PLoS ONE 6(12): e28263. doi:10.1371/journal.pone.0028263

good outcome

bad outcome

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Ac

cu

PS

A(p

g/m

l) L

og

S

ca

le

0.1

1

10

100

1000

Non-Recurrence Recurrence

Oncology

AccuPSATM (Johns Hopkins, NYU)

• Achieved 1,667-fold greater sensitivity than benchmark commercial assays

(LOD = 6 fg/mL). Only test capable of precise measurement of PSA following

prostatectomy.

• Completed pilot study on utility of the assay to predict recurrence of prostate

cancer after radical prostatectomy from a single blood test.

• Convincingly demonstrated that a single AccuPSA result 3-6 months after

surgery highly predictive of cancer recurrence over five year period.

• Clinical value: ability to target secondary

radiation treatment for higher risk group,

identification of ultra-low risk group for whom

such treatment unnecessary

• Commercial leverage: more accurate treatment

targeting; improved efficacy with earlier

treatment; single test result

• Lepor et al., Br. J. Urol. Int., 2011, 12 OCT 2011, DOI:

10.1111/j.1464-410X.2011.10568.x; Wilson et al., Clin. Chem.,

2011, 57, 1712-1721.

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SimoaTM Benefits

Current Offerings Simoa

Current technologies lack sufficient sensitivity for many biomarkers

1000x sensitivity improvement (femtomolar, 10-15)

Limited dynamic range > 4 logs of dynamic range

Time consuming manual process (3-5 hrs.) Fully automated system – sample in, result out, save on FTE cost and manual errors

Imprecision High imprecision of results < 10% CVs thanks to digital approach

Large sample size (25-50 microliters) As little at 1 microliter (with dilution)

Acceptable cost/sample Similar costs to current approach, using conventional reagents

Limited path to Laboratory Developed Tests (LDT) and IVD

Simoa is very amenable to LDT. Partner in IVD is bioMerieux

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Using single molecule measurement Simoa delivers high definition diagnostics. This remarkable digital sensitivity combined with full automation unlocks a world of insight into disease detection, diagnosis, and patient treatment while meeting the demands of today’s laboratory.

Thank You