Atolyzer®A point of care instrument

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A flexible point of care instrument with a disposable cartridge for unlimited assay testing

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The prototype is ready and during 2010 the first heart marker

test will be marketed in Japan.

The immunodiagnostic system represents an entirely new

microfluidic solution and is based on a flexible instrument

and one disposable assay cartridge. This combination is a

mechanical engineering and technological innovation of unique

quality.

The instrument and cartridge are a PoC system characterized

by important features such as flexibility, ultra high sensitivity,

safe and easy handling and low costs.

Atolyzer®

A point of care instrument

During spring 2009 several hundred preclinical heart marker

tests performed by the Atolyzer were carried out at a Danish

hospital and measured against the heart clinics’ own central

laboratory reference tests. Whole blood and plasma samples

run by the Atolyzer correlated 100 %. Likewise, the Atolyzer

test results showed excellent correlation with the ADVIA

Centaur® results.

This brochure introduces six essential engineering elements of

the Atolyzer and cartridge that make the system different from

other PoC devices:

n One system – two parts

n Temperature control

n Blood filtration

n Three step assay

n Anticlustering magnetic particles

n Calibration

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One system – two parts

AtolyzerThe instrument is a platform containing mechanics, electronics

and software only. All essential biomarker information is trans-

ferred from the cartridge to the Atolyzer.

The Atolyzer is controlled by a two level CPU structure: a pro-

prietary CPU hardware driver and standard WIN CE 6.0 CPU

with software. All necessary basic SW and (I/o) LAN, UBS,

Ethernet and Flash are incorporated to connect to HIS

systems.

The instrument is operated by a 3,5” colour touch screen.

The system allows for seamless communication with patient

record data bases and bar code scanning.

The instrument is small (height 6 / width 4 /depth 4.5 inches),

mobile (weight 2.4 lb) and requires no routine maintenance.

The Atolyzer is a flexible instrument able to process any bio-

marker cartridge (to be developed on demand).

The Atolyzer PoC system is constituted of:n The Atolyzer instrument

n One disposable assay cartridge

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Temperature control

CartridgeThe cartridge has a blood inlet (requires 36 µl of blood) and

works without preparatory steps.

Within the cartridge a microfluidic system holds all functionality

necessary to execute and measure the biomarker assay.

Information on chemistry, control of incubation time, tempera-

ture, washing time, algorithm values, cartridge lot number

and calibration values is also stored in the cartridge.

The self-contained reagent system - with waste storage -

leaves the Atolyzer clean after use.

The Atolyzer controls the temperature during assay execution.

This feature is required when adapting the instrument to the

surrounding environment in various climates and settings. The

same feature makes the Atolyzer an unique and exclusive PoC

platform amongst other PoC instruments.

All immunodiagnostic measurements are dependent on tem-

perature control to obtain high sensitivity and low CV values.

The specific biomarker assay determines the temperature,

therefore temperature requirements are transferred from the

cartridge to the Atolyzer.

The temperature control is based on a two way cooling and

heating system. The temperature control system is in direct

contact with the cartridge as well as monitors and controls the

inside temperature of the Atolyzer.

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Basic assay rules set the standard for any comparison betwe-

en PoC testing and central laboratory. These rules define the

incubation time of primary and secondary antibody, require

that assay surface must be large and that mixing and washing

procedures are efficient.

Our assay technology within the cartridge is a self-contained

system based on moving micro magnetic particles between

three locations:

n Two step incubation procedure

n Washing

n Detection

The moving procedure gives high signal / noise ratio due to

four key focus points that set the realm of the engineering

development. The result is a cartridge technology that meet

the required assay standard:

n Large assay surface

n Optimal incubation and shaking of assay surface

n Optimal washing conditions

n No tracer / plasma contamination of detection area

Three step assay

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The transformation of blood into plasma or serum is an in-

evitable requirement in PoC technology. To obtain identical

assay results compared to central laboratory the blood used

in a PoC platform needs to be filtered to same plasma quality

as that of centrifugated blood.

The Atolyzer holds a unique blood filtration unit. The construc-

tion of the filtration system is based on a geometrical proprie-

tary separation membrane.

Liquid sensors located at both ends of the channel monitor

the filtration process.

Key advantagesn Blood filtration time <2 minutes

n Blood volume - 36 µl

n Identical plasma quality compared to centrifugation

Blood filtration

2500

2000

1500

1000

500

0 0 500 1000 1500 2000 2500 Plasma (pg/ml)

Ato

lyze

r B

lood

(p

g/m

l)Atolyzer Blood (pg/ml) versus Atolyzer Plasma (pg/ml)

y = 1.0002x - 17.551R2 = 0.9856

The Blood separation unit

Correlation between whole blood versus plasma results onAtolyzer® (n = 100).The correlation between whole blood and plasma results areshown below. The linear regression is (blood result) = (plasma result) – 17.5 (in pg/ml). R2 is 0.985.

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Atonomics has invented an anticlustering magnetic particle

technology.

The clustering of magnetic particles is a recognized problem

both in central laboratory and PoC instruments and it is also

well known that the clustering is worse in plasma than in

buffer fluids.

In central laboratory analyzers the problem is solved by

violently mixing the fluid and particles, however, this mixing

procedure is not possible in a microfludic system.

Our patent pending microformat technology is a solution that

makes immunoassay testing based on magnetic particles

possible and valid.

HowThe assay principle within the cartridge is based on the mani-

pulation of the magnetic particles. The technology ensures a

homogenous dispersion of the beads during assay processing

thereby enhancing the analyte reaction.

To avoid high CV values because of spontaneously clustering

beads a solution was found to keep the particles separated

during assay execution. The positive effects of this invention are:

n Significantly improved CV values

n Higher sensitivity

n Reduced total assay time

Multi panel detectionThe use of magnetic particles prepares the Atolyzer and

cartridge for single as well as multiplex immunoassay

detection. The Atolyzer technology ensures the separation of

different assays containing magnetic particles before detection

starts.

Magnetic particles

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A mandatory requirement for all in vitro diagnostics systems is

that user measurements can be reproduced and the precision

of the instrument validated. Early and correct treatment is

based on reliable results.

The Atolyzer has a three in one calibration system:

n Liquid quality control

n Test verification

n Assay execution – signal validation

The Atolyzer contains only electronics and mechanics, including

a photo multiplier tube, which makes it a constant instrument.

However, to be able to perform an absolute test of the system

(at optional intervals) a quality control reference liquid is used to

verify that the overall system works and that the value is correct.

When the Atolyzer starts a filtered and constant laser light

emitter checks that all vital functions - including the sensitivity

of the PMT - is working. All mechanical activity will be monito-

red and validated during and after assay execution.

Measurement validationIn a microfluidic system it is critical that the fluid flows as

predicted each time and that the fluid is air bubble free. If not,

treatment risks being based on a false negative value.

The Atolyzer has a built-in electroconductive measurement

system that controls the fluid movement during assay execu-

tion. The system detects whether the fluid is present at the

correct time and whether air bobbles are present in the chan-

nel. Both sample and reagent fluids are monitored and the

shape of the light curve created by the chemistry changes if

the assay is not executed correctly. A reference curve valida-

tes the assay execution.

Calibration

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The Atolyzer is a stand-alone platform due to an integrated

computer.

Computer characteristics n CPU ARM926 EJ 240 MHz

n 32 MB Flash – 64 MB SDRAM

n 3,5” TFT Touch screen – QVGA resolution (320 x 240)

n Ethernet 10/100 MBit

n 1 x USB 2.0 Host

n 1 x USB 2.0 Device

n Windows CE 6.0

Computer options n Windows PC connectivity

n File exchange via standard file explorer

n User interface control by external PC

n LAN network (standard IT infrastructure)

n Keyboard

n Mouse

n Printer

n Webserver interface

n PC / USB key data exchange

n Seamless communication with external records systems

n Bar code scanning

Extra features

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Performance Indicator Specification Current Results Sensitivity/Lower Detection Limit n < 5 pg/ml n Analytical sensitivity < 300 fg/mlMeasuring Range & its Linearity n 104 (1 - 10,000 units) n BNP assay has measuring range of 0.3 to 2,000 pg/ml Reproducibility CV values: CV values:

n high range < 5 % n very high range (> 1000 pg/ml) 1 - 3 % n mid range 5 - 10 % n high range (300 - 1,000 pg/ml) < 8 % n low range 10 - 15 % n mid range (> 40 - 300 pg/ml ) < 8 %

n low range (10 - 40 pg/ml ) < 8 % n very low range (< 10 pg/ml ) < 15 %

Sample Volume n 50 µl n 36 µl whole blood sample volumen 4.6 µl plasma used for the assay after blood separation

Turn-Around-TimeTime from sampling to result n 10 to 15 minutes n 18 minutes Signal/Background n As good as possible pg/ml BNP signal/background

0 1 x 4 28 x

10 116 x40 1,074 x

150 4,850 x600 19,895 x

2,000 34,643 x Correlation with Reference Method n Good correlation with results from n Correlation data with whole blood and plasma samples

standard lab immunoassay system at Atolyzer® with plasma samples at ADVIA Centaur® n Similar CVs as reference lab system

n R2 = 0.97 for Atolyzer® vs. Centaur® for plasma and whole bloodn R2 = 0.99 for plasma and whole blood at Atolyzer® n Similar precision data

Sample Material n Finger-prick/capillary and n Blood separation unit integrated/works reliable venous whole blood n Additional whole blood results (EDTA stabilized blood) generated

Summary of results

Atonomics A/S · Vestre Teglgade 10 · DK-2450 Copenhagen SV · Denmark

www.atonomics.com