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The Use of Gold Nanoparticles in Imaging and Targeted Drug Delivery

The Use of Gold Nanoparticles in Imaging and Targeted Drug Delivery. Simona Dostálová 31. 7. 2015. Investice do rozvoje vzdělávání. Gold nanoparticles. EPR effect & biocompatibility High surface area Easy to produce

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The Use of Gold Nanoparticles in Imaging and Targeted Drug Delivery

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  1. The Use of Gold Nanoparticles in Imaging and Targeted Drug Delivery Simona Dostálová 31. 7. 2015 Investice do rozvoje vzdělávání

  2. Gold nanoparticles • EPR effect & biocompatibility • High surface area • Easy to produce • Unique optical properties (strong absorption, scattering in the visible-near infrared region) • ↓ size = ↑ absorption; ↑ size = ↑ scattering • Chemically inert • High electron density • Spheres/rods/shells/cages/cubes

  3. GNPs in theranostics and biomedicine • Light absorption in „water window“ (700-1200 nm) • Chemical sensors/immunosensors/biosensor • Imaging agent • Photodynamic therapy • Drug delivery vehicles • siRNA, miRNA delivery • Tumor targeting

  4. The past Characterizationofgoldnanoparticles-modifiedCdTequantumdots by scanningelectronmicroscopy NanoCon 2013 3D-printed chipfordetectionofmethicillin-resistantStaphylococcus aureuslabeledwithgoldnanoparticles Electrophoresis Apoferritin as a targeteddeliverysystem MendelNet 2013

  5. The present • Significantchangesofmetallotioneinexpression in tumors => Metal nanoparticle-based immunohistochemistry = immunodetection of metallothionein in tissue using gold-labeled antibodies and LA-ICP-MS

  6. GNPs • Tetrachloroauric acid HAuCl4 1mM[0.0197 g/50 mL] 10 mL • Sodium citrate [0.265 g/10 mL] 0.25 mL stirring Heating in differentconditions Microwavereactionsystem Shaking Power300 W Ramp10:00 Hold 12:00 IR 50°C Power300 W Ramp10:00 Hold 05:00 IR 100°C Power300 W Ramp 05:00 Hold 05:00 IR 120°C SIZE: 2 nm – 40 nm

  7. GNPs conjugation with HWR peptide L 1 2 3 4 + 1.25 µL HWR + 2.50 µL HWR + 5 µL HWR HWR peptide Gold NPs 1= HWR 1 mg/mL 2= GNPs+ 1.25 µL HWR 3= GNPs + 2.50 µL HWR 4= GNPs + 5.00 µL HWR

  8. Conjugation with antibodies GoldNPs + HWR + antibody 1:10000 GoldNPs Gold NPs GoldNPs + HWR GoldNPs + HWR + antibody 1:100 GoldNPs + HWR + antibody 1:1000 L 1 2 3 4 5 6 7 1= HWR 1:100 2= IgG 1:100 3= GNPs 4= GNPs + HWR 5= GNPs + HWR + IgG 1:100 6= GNPs + HWR + IgG 1:1000 7= GNPs + HWR + IgG 1:1000 Too low sensitivity => optimization on QDs

  9. QDs conjugation with HWR L= Ladder 1= HWR [1mg/mL] 2= QDs 110°C precipitated 3= QDs 110°C + HWR [0.0208 mg/mL] Peptide concentration [0.0208 mg/mL] causes the saturation of available bonds on QDs.

  10. QDs conjugation with HWR and antibodies L= Ladder 1= HWR [1mg/mL] 2= IgY [54 mg/mL] 3= QDs 110°C precipitated 4= QDs 110°C + HWR [0.0208 mg/mL] 5= QDs + HWR + IgY [13.675 mg/mL] Theconditionsof gel electrophoresiswereoptimized: - running buffer TrisTricinewith pH adjusted to 9, - loadingbufferwithmercaptoethanol (stabilizing agent) Optimalratiosof HWR and antibodywerefound L 1 2 3 4 5

  11. ELISA

  12. Dot blot LA-ICP-MS

  13. Conclusion • Gold nanoparticles have many uses in theranostics and biomedicine • One of them - metal nanoparticle-based immunohistochemistry • We optimized the binding of HWR peptide and antibodies • We need to optimize the HWR-GNPs binding and detection of complex

  14. Acknowledgements Bc. Monica Vazzana Mgr. Marketa Vaculovicova, Ph.D. Doc. RNDr. Pavel Kopel, Ph.D. Prof. Ing. Rene Kizek, Ph.D. Investice do rozvoje vzdělávání 11

  15. Thank you for your attention Investice do rozvoje vzdělávání

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