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EBS2019 Florence, Italy, February 18-21, 2019 Microarray based - PowerPoint PPT Presentation

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2 European Biosensor Symposium 2019 EBS2019 Florence, Italy, February 18-21, 2019 Microarray based platform for extracellular vesicles imaging and phenotyping D. Brambilla, M. Cretich, A. Romanato, M. Odinolfi, E. Chiodi, S. Descroix, A.

  1. 2° European Biosensor Symposium 2019 EBS2019 Florence, Italy, February 18-21, 2019

  2. Microarray based platform for extracellular vesicles imaging and phenotyping D. Brambilla, M. Cretich, A. Romanato, M. Odinolfi, E. Chiodi, S. Descroix, A. Yalcin Ozkumur, M.S. Unlu, M. Chiari

  3. ü Microarray platform for Exosome Immune-Capture ü Interferometric detection of Single NanoParticles

  4. Extracellular Vesicles Exosomes q Early Endosome q Multi Vesicular Bodies q Lysosomial degradation q Exosome Release “Extracellular vesicles: Exosomes, microvesicles, and friends” G. Raposo and W. Stoorvogel - J. Cell Biol. Vol. 200 No. 4 373–383

  5. Exosome: Diagnostic potential q Mirrors the cell of origin q Liquid Biopsy ü Cancer ü Neurodegenerative diseases

  6. Exosome: Characteristics ü Number ü Size distribution ü Membrane composition ü Cargo

  7. Exosome: Capture ü Microarray platform for Exosome Immune-Capture

  8. Microarray technique

  9. Polymeric Coating Copoly(DMA-NAS-MAPS) m=97 n=2 p=1

  10. Probe Immobilization Antibodies (proteic probes) ü Tetraspanins (CD9, CD63)

  11. DNA-directed Immobilization #1

  12. DNA-directed Immobilization #2 ü DNA Spot morphology ü Antibody specificity ü Improved flexibility

  13. Selectivity ü Multiplexing

  14. Detection ü Interferometric detection of Single NanoParticles

  15. SP-IRIS: Sizing and Phenotyping #1 ü IRIS: Interferometric Reflectance Imaging System § Led-based technique § Matter immobilized on surface § Shifts of reflectivity Image contrast G. G. Daaboul et al. (2011) “LED-based Interferometric Reflectance Imaging Sensor for quantitative dynamic monitoring of biomolecular interactions” Biosensors and Bioelectronics, 26, 5, 2221-2227.

  16. SP-IRIS: Sizing and Phenotyping #2 G. G. Daaboul et al. (2016) “Digital Detection of Exosomes by Interferometric Imaging” Scientific Reports, 6:37246.

  17. SP-IRIS: Sizing and Phenotyping #3 ü Digital count of nanoparticles (50-200 nm) ü Size distribution ü Phenotyping (microarray platform) G. G. Daaboul et al. (2016) “Digital Detection of Exosomes by Interferometric Imaging” Scientific Reports, 6:37246.

  18. Exosome Immune-capture: Results #1 Spot : DNA-directed antiCD9 Incubation : Human Blood Serum, 2.5h, RT

  19. Exosome Immune-capture: Results #2 Spot : DNA-directed antiCD9 Incubation : Human Blood Serum, 2.5h, RT

  20. Exosome Immune-capture: Results #3 Spot : DNA-directed antiCD9 Incubation : Human Blood Serum, 2.5h, RT

  21. Exosome Immune-capture: Results #4

  22. Exosome Immune-capture: Results #5

  23. Exosome Immune-capture: Results #6 unmodified AntiCD63 DNA-Directed AntiCD63

  24. Next Step: Real Time SP-IRIS ü Kd constant Affinity ü Distinguish specific and non-specific events

  25. Acknowledgments FET-Open 2016-2017 Grant Agreement no. 766466

  26. THANK YOU FOR THE KIND ATTENTION

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