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STCU-NATO Workshop "From Science to Business"STCU-NATO Workshop "From Science to Business" Kyiv — October 11–12, 2006Kyiv — October 11–12, 2006

Tel.: +380 (57) 341-02-72Tel.: +380 (57) 341-02-72E-mail: patsenker@isc.kharkov.com E-mail: patsenker@isc.kharkov.com http://www.isc.kharkov.com/oldhttp://www.isc.kharkov.com/old

Leonid PatsenkerLeonid PatsenkerPh.D., Head of DepartmentPh.D., Head of Department

Fluorescent Probes and Labels Fluorescent Probes and Labels for Biomedical Applicationsfor Biomedical Applications

National Academy of Sciences of UkraineNational Academy of Sciences of UkraineState Scientific Institution State Scientific Institution

"Institute for Single Crystals""Institute for Single Crystals"Department of Organic Luminophores and DyesDepartment of Organic Luminophores and Dyes 60 Lenin Ave., Kharkov, 61001, Ukraine60 Lenin Ave., Kharkov, 61001, Ukraine

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Talk outlineTalk outline

1.1. What we offerWhat we offer: brief technology description : brief technology description

2.2. StageStage of development of development

3.3. The The marketmarket for fluorescent probes and labels for fluorescent probes and labels

4.4. WhoWho needs it? needs it?

5.5. AdvantagesAdvantages of our materials of our materials

6.6. Our Our competitorscompetitors and how to and how to beatbeat them them

7.7. OpportunityOpportunity for joint work for joint work

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Proprietary information statementProprietary information statement

The materials presented in this talk are The materials presented in this talk are available for licensingavailable for licensing and and joint product joint product developmentdevelopment

NoneNone of the slides contain any confidential or of the slides contain any confidential or proprietary informationproprietary information

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What We OfferWhat We Offer

Fluorescent DyesFluorescent Dyes((LuminophoresLuminophores))

Non-fluorescent DyesNon-fluorescent Dyes((QuenchersQuenchers))

Dyes for Biomedical ApplicationsDyes for Biomedical Applications

Reactive MarkersReactive Markers((LabelsLabels))

Non-reactive Non-reactive MarkersMarkers((ProbesProbes))

ClassificationClassificationDyes for Dyes for

MicrospheresMicrospheres

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We Offer the Life Sciences Related We Offer the Life Sciences Related Products and TechnologiesProducts and Technologies

• Fluorescent probesFluorescent probes and and labelslabels but alsobut also

• Non-fluorescent quenchersNon-fluorescent quenchers for biological and for biological and pharmaceutical researches, and biomedical assays pharmaceutical researches, and biomedical assays

• Classification dyesClassification dyes for single or multiple encoding of for single or multiple encoding of microspheres used in High-throughput Screening (HTS)microspheres used in High-throughput Screening (HTS)

• TechnologiesTechnologies to synthesize, produce and use these materials to synthesize, produce and use these materials

• Custom synthesis, R&DCustom synthesis, R&D

100 products100 products

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These materials and technologies These materials and technologies are available for licensingare available for licensing

Fluorescent probes, labels, and Fluorescent probes, labels, and non-fluorescent dyes non-fluorescent dyes (quenchers)(quenchers) for biological and for biological and biomedical applications, biomedical applications, pharmacology and pharmacology and environmental pollution environmental pollution monitoringmonitoring

Classification dyesClassification dyes for beads for beads

Other luminescence-based Other luminescence-based materialsmaterials

TechnologiesTechnologies to synthesize, to synthesize, produce and use these produce and use these materialsmaterials

Some of Our Existed Patents and Some of Our Existed Patents and Patent Applications: Patent Applications:

1.1. PCT WO 2003/087052 A3 (2003) PCT WO 2003/087052 A3 (2003)

2.2. US Patent 6538129 (2003)US Patent 6538129 (2003)

3.3. US Patent Application US Patent Application 2004/0166515 A1 (2004)2004/0166515 A1 (2004)

4.4. US Patent Application US Patent Application 2003/0235846 A1 (2003)2003/0235846 A1 (2003)

5.5. US Patent ApplicationUS Patent Application 20052005/0/0202565202565 A1 (2005) A1 (2005)

Stage of DevelopmentStage of Development

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A Sample Page @ Online Product CatalogA Sample Page @ Online Product Cataloghttp://www.isc.kharkov.com/oldhttp://www.isc.kharkov.com/oldhttp://www.isc.kharkov.com/oldhttp://www.isc.kharkov.com/old

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MARKET for Fluorescent Probes & Labels:MARKET for Fluorescent Probes & Labels: Biology, Medicine, Pharmacology, and Biology, Medicine, Pharmacology, and

Environmental Pollution MonitoringEnvironmental Pollution Monitoring

• Biological Imaging• Clinical Diagnostics• High-throughput Screening (HTS)• Real-time PCR Based Applications• Cellular and Molecular Biology• Cytology• Immunology• Genomics• Proteomics• Enzymatic • Drug Screening• Photodynamic Therapy• Environmental (chemical and

bacterial) pollution monitoring

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Who needs our markers?Who needs our markers?The potential application of fluorescent probes, labels, and The potential application of fluorescent probes, labels, and classification dyes is very versatile.classification dyes is very versatile. Therefore Therefore the market for these the market for these materials is hugematerials is huge

Any company using fluorescenceAny company using fluorescence as a read-out parameter can be as a read-out parameter can be interested in our productsinterested in our products

These are organizations working These are organizations working in the life-science marketin the life-science market: :

Biomedical companies Biomedical companies

Clinical laboratoriesClinical laboratories

Environmental monitoring laboratories Environmental monitoring laboratories

Academic researchers in the biological, chemical and medical Academic researchers in the biological, chemical and medical fieldsfields

Fluorescent dyes for biomedical applications Fluorescent dyes for biomedical applications are a multi-billion dollar businessare a multi-billion dollar business!!

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Our products Our products have been tested at:have been tested at:

SETA Biomedicals (USA)SETA Biomedicals (USA) Perkin Elmer, Inc. (USA)Perkin Elmer, Inc. (USA) Tecan (Austria)Tecan (Austria) ReaMetrix, Inc. (USA)ReaMetrix, Inc. (USA) Dako (Denmark)Dako (Denmark) Luminex Corp. (USA) Luminex Corp. (USA) LJL Biosystems (USA)LJL Biosystems (USA) Molecular Devices (USA)Molecular Devices (USA) ISS (USA)ISS (USA) Avanti Polar Lipids, Inc. Avanti Polar Lipids, Inc.

(USA)(USA) Edinburgh Instruments (Great Edinburgh Instruments (Great

Britain)Britain) Becton Dickinson (USA)Becton Dickinson (USA) Pierce Biotechnology (USA)Pierce Biotechnology (USA) Glen Research Corp. (USA)Glen Research Corp. (USA)

Potential customers:Potential customers:

Abbot (USA)Abbot (USA) Bayer (Germany) Bayer (Germany) Pfizer (USA)Pfizer (USA) GE Healthcare (Amersham) (USA)GE Healthcare (Amersham) (USA) Invitrogen (Molecular Probes) (USA)Invitrogen (Molecular Probes) (USA) Glaxo Smith Kline (USA) Glaxo Smith Kline (USA) CIS Bio International (France)CIS Bio International (France) Alpha Diagnostics International (USA)Alpha Diagnostics International (USA) Dakota Technologies, Inc. (USA)Dakota Technologies, Inc. (USA) IQ Products (The Netherlands)IQ Products (The Netherlands) GeneFocus (Biomedical Photometrics GeneFocus (Biomedical Photometrics

Inc.) (Canada)Inc.) (Canada) Many others ...Many others ...

1111

300 400 500 600 700 8000.0

0.2

0.4

0.6

0.8

1.0

1.2

Square-755-di-NHSSeta-670-NH-di-NHS

Square-565-mono-NHS

Seta-635-B-di-NHS

Cy5

Abs

orba

nce

[a.u

.]

Wavelenght [nm]

We offer biomedical markers for We offer biomedical markers for a wide spectral range, a wide spectral range, from the ultra-violet to the near-infraredfrom the ultra-violet to the near-infrared

As compared to materials from other As compared to materials from other producers our markers have certain producers our markers have certain

advantages in most all characteristicsadvantages in most all characteristics

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300 400 500 600 7000.0

0.2

0.4

0.6

0.8

1.0

1.2

Absorptionin blue region

Seta-670-NH-di-NHSSquare-565-mono-NHS

Cy5

Ab

sorb

ance

[a.

u.]

Wavelenght [nm]

New markers are excitable not New markers are excitable not only with only with red diode lasersred diode lasers but also but also

with with blue diode lasersblue diode lasers, , LEDsLEDs, , and and UV lightUV light

650 700 750 800 850 9000

10

20

30

40

50

60

70Square-670 +10

–4 M BSA

Square-670

Flu

ore

scen

ce In

tetc

ity

[a.u

.]

Wavelength [nm]

New markers New markers show noticeableshow noticeable increase of fluorescence intensity increase of fluorescence intensity and and longer fluorescence lifetimeslonger fluorescence lifetimes

in presence of proteinin presence of protein

Advantages of Our MarkersAdvantages of Our MarkersCompatibility with VariousCompatibility with VariousExcitation Light SourcesExcitation Light Sources

Response for Response for MicroenvironmentMicroenvironment

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New reactive labels demonstrate New reactive labels demonstrate higher brightnesshigher brightness when bound to when bound to protein as compared to e.g. Cy5 protein as compared to e.g. Cy5

of of GE HealthcareGE Healthcare

0 1 2 30

20

40

60

Seta-635-B — BSA conjugates

Cy5—BSA conjugates

Seta-635 — BSA conjugates

Dye-to-Protein Ratio

Qu

an

tum

Yie

ld [

%]

400 500 600 7000,00

0,02

0,04

0,06

pH 11.88

Seta-660-B-NH

pH 5.46

Wavelength [nm]

Ab

so

rba

nc

e

New markers can be made New markers can be made pH-sensitivepH-sensitive

Advantages of Our MarkersAdvantages of Our Markers

High BrightnessHigh Brightness pH SensitivitypH Sensitivity

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Decay of brightness Decay of brightness of a biological imageof a biological image

High PhotostabilityHigh Photostability

0 10 20 30 40 50 600.94

0.96

0.98

1.00

1.02

Seta-635-B (ISC)

Seta-635-S (ISC)

Cy5 (GE Healthcare)

Rel

ativ

e B

rig

htn

ess

Time [sec]

Biological ImagingBiological Imaging

Advantages of Our MarkersAdvantages of Our Markers

New markers are New markers are much more photostablemuch more photostable compared to those from other compared to those from other producers, which is particularly important to obtain higher-stable producers, which is particularly important to obtain higher-stable

biological imagesbiological images

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400 500 600 700 800 900 10000

20000

40000

60000

80000

100000

120000 Excitation

Fluorescence

Ext

inct

ion

Co

effi

cien

t [M

–1cm

–1]

Wavelength [nm]300 400 500 600 700 800 900 1000

0.0

0.2

0.4

0.6

0.8

1.0

NoFluorescence

Excitation

Ab

sorb

ance

[a.

u.]

Wavelenght [nm]

Advantages of Our MarkersAdvantages of Our MarkersFluorescent Probes Fluorescent Probes and Reactive Labelsand Reactive Labels

Effective Quenchers Effective Quenchers (Real Dark Dyes)(Real Dark Dyes)

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CompetitionCompetition

InvitrogenInvitrogen (Molecular Probes, USA)(Molecular Probes, USA)GE HealthcareGE Healthcare (Amersham(Amersham, , USAUSA))

Leading Competitors:Leading Competitors:

Dyomics GmbHDyomics GmbH (Germany)(Germany)

Active Motif Chromeon GmbHActive Motif Chromeon GmbH (Germany)(Germany)

Several small companiesSeveral small companies

Other Competitors:Other Competitors:

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Our biomedical materials have Our biomedical materials have better characteristicsbetter characteristics::

Higher brightness Higher brightness

Longer luminescence lifetimes, which make them useful Longer luminescence lifetimes, which make them useful for the FLT and polarization based assaysfor the FLT and polarization based assays

Higher photostabilityHigher photostability

Our materials are Our materials are cheapercheaper

How will our materials How will our materials beat the competitors?beat the competitors?

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Competitive MatrixCompetitive Matrix

CharacteristicCharacteristicOur MarkerOur Marker

Seta-640-di-NHSSeta-640-di-NHS

InvitrogenInvitrogen

(Molecular Probes(Molecular Probes))

Alexa Fluor 647Alexa Fluor 647

GE HealthcareGE Healthcare

((AmershamAmersham))

Cy5 NHS esterCy5 NHS ester

Absorption / Emission Wavelength 648 / 656 nm648 / 656 nm 650650 // 668 nm668 nm 648 / 664 nm648 / 664 nm

Quantum Yieldof BSA Conjugate 65%65% 42%42% 25%25%

Luminescence Lifetime (Free / Bound to BSA) 0.7 / 3.0 ns0.7 / 3.0 ns 1.1 / 1.2 ns1.1 / 1.2 ns 1.0 / 1.2 ns1.0 / 1.2 ns

Applicability for the polarization

and FLT assaysYesYes NoNo NoNo

Photostability 90%90% 80%80% 60%60%

Price$125 / 1 mg $125 / 1 mg

$2,500 / 50 mg$2,500 / 50 mg

$212 / 1 mg $212 / 1 mg

$4,300 / 50 mg$4,300 / 50 mg

$222 / 1 mg $222 / 1 mg

$4,455 / 50 mg$4,455 / 50 mg

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Competitive MatrixCompetitive Matrix

CharacteristicCharacteristicOur MarkerOur Marker

SeTau-380-SeTau-380-NHSNHS

Our MarkerOur Marker

SeTau-425-SeTau-425-NHSNHS

InvitrogenInvitrogen

DPHDPH

InvitrogenInvitrogen

NBDNBD

Absorption / Emission Wavelength 381 / 480 nm381 / 480 nm 425 / 545 nm425 / 545 nm 360 / 430 nm360 / 430 nm 470 / 530 nm470 / 530 nm

Quantum Yield (Free / Bound to BSA) 56% / 44%56% / 44% 39% / 16%39% / 16% 0% / 80%0% / 80% 0% / 32%0% / 32%

Fluorescence in water YesYes YesYes NoNo NoNo

Luminescence Lifetime (Free / Bound to BSA) 33 / 31 ns33 / 31 ns 26 / 13 ns26 / 13 ns 4 / 8 ns4 / 8 ns 5 / 10 ns5 / 10 ns

Price per 1 mg $70$70 $80$80 $78$78 $180$180

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Opportunities for Joint WorkOpportunities for Joint Work

Search for Search for the continuous customersthe continuous customers of our products and of our products and technologies ...technologies ...

Collaboration with companies which are interested in Collaboration with companies which are interested in further further testingtesting, , licensinglicensing and and commercializationcommercialization of our of our materials and technologies ...materials and technologies ...

Collaboration with companies which are interested in Collaboration with companies which are interested in further joint development of our materialsfurther joint development of our materials towards making towards making of new of new "Marker — Specific Biological Species""Marker — Specific Biological Species" conjugates conjugates such as fluorescently labeled peptides, oligonucleotides, such as fluorescently labeled peptides, oligonucleotides, proteins, antibodies, and antigens but also proteins, antibodies, and antigens but also development of development of kits and assayskits and assays based on our markers ... based on our markers ...

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STCU Project U111STCU Project U111 Red and NIR Absorbing and Emitting Fluorescent Probes and Labels

for Use in Biological and Biomedical Research

2004 — 2006

CommercializationCommercialization

STCU 3804STCU 3804Fluorescence Lifetime-based Probes

and Labels for Biomedical Assays and Bacterial Pollution Monitoring

Submitted

STCU Project 3795STCU Project 3795 Improving Multiplexed Analysis

Systems for Environmental Pollution Monitoring and Medical Diagnostics

Using Novel Classification Dyes

Submitted

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Leonid PatsenkerLeonid PatsenkerPh.D., Head of DepartmentPh.D., Head of Department

Tel.: +38 (057) 341-02-72 Tel.: +38 (057) 341-02-72 Fax: +38 (057) 340-93-43 Fax: +38 (057) 340-93-43

E-mail: patsenker@isc.kharkov.com E-mail: patsenker@isc.kharkov.com http://www.isc.kharkov.comhttp://www.isc.kharkov.com

National Academy of Sciences of UkraineNational Academy of Sciences of Ukraine

State Scientific Institution State Scientific Institution "Institute for Single Crystals""Institute for Single Crystals"Department of Organic Luminophores and Dyes Department of Organic Luminophores and Dyes 60 Lenin Ave., Kharkov, 61001, Ukraine60 Lenin Ave., Kharkov, 61001, Ukraine

Contact InformationContact Information