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Jacqueline Vet   24 mei 2012
2007

                        ZZP-organisation



 Activities:
     • design and development of prototype molecular diagnostic assays

       • workshops/ courses




  Business model: contract research
ORGANISATION                                                    ZZP




                                               NYtor
                                           Universitair Bedrijvencentrum
                                           Toernooiveld 102
                                           Nijmegen




                                                Molecular Beacon Lab (USA)                 Bio-Rad
      HAN BioCentre                             PHRI, UMDNJ, Dept. of Molecular Genetics   1.   Apparatus
                                                                                           2.   Reagents
                                                1.     Assay design                        3.   Network
1.   Collaboration agreement
                                                2.     New technological developments
2.    Use labspace                                                                         Biolegio
                                                3.     Collaboration projects              1.   primers-probe
3.   NYtor hires research analist                                                               synthesis
4.   Use both networks for new customers




     www.hanbiocentre.nl
                                                     www.molecular-beacons.org
2012
                                      BV



                   SERVICES                            DEVELOPMENT




Business model: designed assays are licensed to suitable party : fee +royalties




Activities: own product development
ORGANISATION                                    BV



     NYtor office                          NYtor laboratory
     Universitair Bedrijvencentrum         Mercator III
     Toernooiveld 102                      Toernooiveld 1
     Nijmegen                              Nijmegen




Molecular Beacon Lab (USA)                                  Bio-Rad
PHRI, UMDNJ, Dept. of Molecular Genetics                    1.   Apparatus
                                                            2.   Reagents
1.    Assay design                                          3.   Network

2.    New technological developments
                                                            Biolegio
3.    Collaboration projects                                1.   primers-probe
                                                                 synthesis




        www.molecular-beacons.org
NYtor TECHNOLOGY
NYtor TECHNOLOGY




     Molecular beacon qPCR-technology

     =

     DNA-amplification + high specific DNA-detection
              (target)                 (molecular beacon)




 24-05-2012
NYtor TECHNOLOGY - II




              Multiplex Four Target PCR Assay




 24-05-2012
NYtor TECHNOLOGY - III



   Technique developed at Public Health Research Institute (NY, VS)
   1995-1998, postdoctoral fellowship: co-inventor / co-patent holder




Licensees: > 50 e.g.: bioMerieux, Becton Dickinson, Gen-Probe, Abbott Laboratories,
                        Bio-Rad, Affymetrix, Stratagene, Cepheid




     24-05-2012
Applications




Fields of use for the molecular beacons technology
1. Human in vitro diagnostics
a. Testing for infectious agents   7. Agricultural infectious agents

b. Genetic testing                 8. Forensics

c. Oncology                        9. Paternity testing

2. Research                        10. High-density probe arrays

3. GMO testing                     11. Industrial microbiology

4. Food testing                    12. Blood products quality control

5. Veterinary diagnostics          13. Environmental pathogens

6. Hybrid seed production
Overview developed qPCR assays - I

FOOD/WATER:
   •Legionella pneumophila
   •Legionella species
   •Campylobacter jejuni-coli-lari multiplex
   •E.coli 0157
   •Listeria monocytogenes
   •Listeria species



PHARMACEUTICAL:
   •CHO qPCR: quantitation of host-cell residual DNA from Chinese Hamster Ovary (CHO) cells




VETERINARY:
   •Scrapie genotyping: multiplex detection 5 SNPs (sheep)
   •Helicobacter ganmani-rodentium multiplex (fish)




     24-05-2012
Overview developed qPCR assays - II

CLINICAL:
   •HIV-1, HIV-2, HTLV-II and HTLV-II multiplex
   •Chlamydia pneumonia and Mycoplasma pneumonia multiplex
   •Clostridium difficile ribotypes 017 and 027 multiplex
   •Methylenetetrahydrofolate reductase (MTHFR)-gene (cardiovascular disease)



MISCELLANEOUS
   •RT-qPCR assay to dermine optimum moment of harvesting (pears)
   •Lactococcus lactis spp. lactis, spp. cremoris, spp. lactis biovar diacetylactis and
    ,Leuconostoc monoplex (starter cultures yogurt and cheese)




     24-05-2012
PROJECTS                     BV




 1.      Superbug project (EFRO) - Check-Points

 2.      Veterinary diagnostics - Biochek

 3.      Personalized Medicine - Future Diagnostics




 24-05-2012
SUPERBUG PROJECT


- Fast increase multi resistant bacteria (superbugs)

- Cause resistance: enzymes destroy antibiotics
   • Extended Spectrum Beta-Lactamases (ESBL)
   • Klebsiella Pneumonia Carbapenemase (KPC)
   • New Delhi Metallo-Beta-Lactamase (NDM-1)

    - ESBLs in Klebsiella pneumoniae




            2002                   2005                      2008
      Bron: http://ecdc.europa.eu/en/activities/surveillance/EARS-Net/Pages/Database.aspx




          Crucial: identification of bacteria at early stage

           •      optimal treatment of patient
           •      timely detection outbreaks  prevent spread



     24-05-2012
PRODUCT DEVELOPMENT


       SuperBug assay

       Rapid test for detection of
       multi resistant bacteria




                        DNA                   PROBE-LIGATIE            MICROARRAY
conventionele        EXTRACTIE                  INCUBATIE          INCUBATIE & ANALYSE
    test                 1 uur                   3,5 uur                 2,5 uur


 SuperBug               DNA       qPCR
                     EXTRACTIE   met MB
   assay
                         1 uur    1 uur



                    0                     2                                              7
                                                    Tijd (uren)


                 Essential time saving ! (5 hour reduction to detection)



            24-05-2012
SAMENWERKINGSPARTNER




Bedrijfsstrategie NYtor: ontwikkelde testen worden ter licentie aangeboden
aan een geschikte marktpartij met bijbehorende royaltyregeling.



Voor de SuperBug assay samenwerking met:

Check-Points BV (Wageningen)

• wereldwijde marktleider op gebied van analyse van
  gramnegatieve bacteriën.
• internationaal distributienetwerk in 30 landen




NYtor + Check-Points: sterk team  snelle en efficiënte onwikkeling van
de superbug assay




      24-05-2011
SUPERBUG – INVESTMENT/EARNINGS




                                                    %
       EFRO grant                                   50
       Upfront Fee                                  25
       NYtor/KERN                                   25
       Total                                       100




 700
 600       SuperBug assay – 10 year prognosis
 500
 400
                                                                              REALISTIC
 300
 200                                                                          WORST CASE
 100                                                                          OPTIMISTIC
   0
-100   2011    2012   2013   2014   2015   2016   2017   2018   2019   2020
-200




        24-05-2011
‘PERSONALIZED MEDICINE’-ASSAYS


•   accurate diagnostic tests that identify patients who can benefit from
    targeted therapies




•   Strategic collaboration with Future Diagnostics


   NYtor: prototype assay development

   Future diagnostics: validation of the prototype assays
FUTURE PERSPECTIVES



•   Lab-on-a-Chip: coupling self-reporting molecular beacons to micro array format




•   Sloppy molecular beacons: increase multiplex format of qPCR up to hunderd targets
FUTURE CHALLENGES




   •     new facilities
   •     NYtor team: find right people
   •     financing
   •     business strategy

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Molecular Diagnostics: guided by the light

  • 1. Jacqueline Vet 24 mei 2012
  • 2.
  • 3. 2007 ZZP-organisation Activities: • design and development of prototype molecular diagnostic assays • workshops/ courses Business model: contract research
  • 4. ORGANISATION ZZP NYtor Universitair Bedrijvencentrum Toernooiveld 102 Nijmegen Molecular Beacon Lab (USA) Bio-Rad HAN BioCentre PHRI, UMDNJ, Dept. of Molecular Genetics 1. Apparatus 2. Reagents 1. Assay design 3. Network 1. Collaboration agreement 2. New technological developments 2. Use labspace Biolegio 3. Collaboration projects 1. primers-probe 3. NYtor hires research analist synthesis 4. Use both networks for new customers www.hanbiocentre.nl www.molecular-beacons.org
  • 5. 2012 BV SERVICES DEVELOPMENT Business model: designed assays are licensed to suitable party : fee +royalties Activities: own product development
  • 6. ORGANISATION BV NYtor office NYtor laboratory Universitair Bedrijvencentrum Mercator III Toernooiveld 102 Toernooiveld 1 Nijmegen Nijmegen Molecular Beacon Lab (USA) Bio-Rad PHRI, UMDNJ, Dept. of Molecular Genetics 1. Apparatus 2. Reagents 1. Assay design 3. Network 2. New technological developments Biolegio 3. Collaboration projects 1. primers-probe synthesis www.molecular-beacons.org
  • 8. NYtor TECHNOLOGY Molecular beacon qPCR-technology = DNA-amplification + high specific DNA-detection (target) (molecular beacon) 24-05-2012
  • 9. NYtor TECHNOLOGY - II Multiplex Four Target PCR Assay 24-05-2012
  • 10. NYtor TECHNOLOGY - III Technique developed at Public Health Research Institute (NY, VS) 1995-1998, postdoctoral fellowship: co-inventor / co-patent holder Licensees: > 50 e.g.: bioMerieux, Becton Dickinson, Gen-Probe, Abbott Laboratories, Bio-Rad, Affymetrix, Stratagene, Cepheid 24-05-2012
  • 11. Applications Fields of use for the molecular beacons technology 1. Human in vitro diagnostics a. Testing for infectious agents 7. Agricultural infectious agents b. Genetic testing 8. Forensics c. Oncology 9. Paternity testing 2. Research 10. High-density probe arrays 3. GMO testing 11. Industrial microbiology 4. Food testing 12. Blood products quality control 5. Veterinary diagnostics 13. Environmental pathogens 6. Hybrid seed production
  • 12. Overview developed qPCR assays - I FOOD/WATER: •Legionella pneumophila •Legionella species •Campylobacter jejuni-coli-lari multiplex •E.coli 0157 •Listeria monocytogenes •Listeria species PHARMACEUTICAL: •CHO qPCR: quantitation of host-cell residual DNA from Chinese Hamster Ovary (CHO) cells VETERINARY: •Scrapie genotyping: multiplex detection 5 SNPs (sheep) •Helicobacter ganmani-rodentium multiplex (fish) 24-05-2012
  • 13. Overview developed qPCR assays - II CLINICAL: •HIV-1, HIV-2, HTLV-II and HTLV-II multiplex •Chlamydia pneumonia and Mycoplasma pneumonia multiplex •Clostridium difficile ribotypes 017 and 027 multiplex •Methylenetetrahydrofolate reductase (MTHFR)-gene (cardiovascular disease) MISCELLANEOUS •RT-qPCR assay to dermine optimum moment of harvesting (pears) •Lactococcus lactis spp. lactis, spp. cremoris, spp. lactis biovar diacetylactis and ,Leuconostoc monoplex (starter cultures yogurt and cheese) 24-05-2012
  • 14. PROJECTS BV 1. Superbug project (EFRO) - Check-Points 2. Veterinary diagnostics - Biochek 3. Personalized Medicine - Future Diagnostics 24-05-2012
  • 15. SUPERBUG PROJECT - Fast increase multi resistant bacteria (superbugs) - Cause resistance: enzymes destroy antibiotics • Extended Spectrum Beta-Lactamases (ESBL) • Klebsiella Pneumonia Carbapenemase (KPC) • New Delhi Metallo-Beta-Lactamase (NDM-1) - ESBLs in Klebsiella pneumoniae 2002 2005 2008 Bron: http://ecdc.europa.eu/en/activities/surveillance/EARS-Net/Pages/Database.aspx Crucial: identification of bacteria at early stage • optimal treatment of patient • timely detection outbreaks  prevent spread 24-05-2012
  • 16. PRODUCT DEVELOPMENT SuperBug assay Rapid test for detection of multi resistant bacteria DNA PROBE-LIGATIE MICROARRAY conventionele EXTRACTIE INCUBATIE INCUBATIE & ANALYSE test 1 uur 3,5 uur 2,5 uur SuperBug DNA qPCR EXTRACTIE met MB assay 1 uur 1 uur 0 2 7 Tijd (uren)  Essential time saving ! (5 hour reduction to detection) 24-05-2012
  • 17. SAMENWERKINGSPARTNER Bedrijfsstrategie NYtor: ontwikkelde testen worden ter licentie aangeboden aan een geschikte marktpartij met bijbehorende royaltyregeling. Voor de SuperBug assay samenwerking met: Check-Points BV (Wageningen) • wereldwijde marktleider op gebied van analyse van gramnegatieve bacteriën. • internationaal distributienetwerk in 30 landen NYtor + Check-Points: sterk team  snelle en efficiënte onwikkeling van de superbug assay 24-05-2011
  • 18. SUPERBUG – INVESTMENT/EARNINGS % EFRO grant 50 Upfront Fee 25 NYtor/KERN 25 Total 100 700 600 SuperBug assay – 10 year prognosis 500 400 REALISTIC 300 200 WORST CASE 100 OPTIMISTIC 0 -100 2011 2012 2013 2014 2015 2016 2017 2018 2019 2020 -200 24-05-2011
  • 19. ‘PERSONALIZED MEDICINE’-ASSAYS • accurate diagnostic tests that identify patients who can benefit from targeted therapies • Strategic collaboration with Future Diagnostics  NYtor: prototype assay development  Future diagnostics: validation of the prototype assays
  • 20. FUTURE PERSPECTIVES • Lab-on-a-Chip: coupling self-reporting molecular beacons to micro array format • Sloppy molecular beacons: increase multiplex format of qPCR up to hunderd targets
  • 21. FUTURE CHALLENGES • new facilities • NYtor team: find right people • financing • business strategy