1. The Use of Gold Nanoparticles in Imaging and Targeted Drug Delivery
Simona Dostálová
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“ ( nm)
Chemical sensors/immunosensors/biosensor
Imaging agent
Photodynamic therapy
Drug delivery vehicles
siRNA, miRNA delivery
Tumor targeting

4. The past
Characterization of gold nanoparticles-modified CdTe quantum dots by scanning electron microscopy
NanoCon 2013
3D-printed chip for detection of methicillin-resistant Staphylococcus aureus labeled with gold nanoparticles
Electrophoresis
Apoferritin as a targeted delivery system
MendelNet 2013

5. The present Significant changes of metallotionein expression 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] mL
stirring
Heating in different conditions
Microwave reaction system
Shaking
Power 300 W
Ramp 10:00
Hold 12:00
IR 50°C
Power 300 W
Ramp 10:00
Hold 05:00
IR 100°C
Power 300 W
Ramp 05:00
Hold 05:00
IR 120°C
SIZE: 2 nm – 40 nm

7. GNPs conjugation with HWR peptideL 1 2 3 4
µL HWR
µL HWR
+ 5 µL HWR
HWR peptide
Gold NPs
Added HWR (μl)
Average size (nm) 32
1.25 50
2.50 90
5.00
150
1= HWR 1 mg/mL
2= GNPs µL HWR
3= GNPs µL HWR
4= GNPs µL HWR

8. Conjugation with antibodies
Gold NPs
Gold NPs
Gold NPs +
HWR
Gold NPs +
HWR +
antibody
1:100
Gold NPs +
HWR +
antibody
1:1000
Gold NPs +
HWR +
antibody
1:10000 L
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 [ mg/mL]
Peptide concentration [ 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 [ mg/mL]
5= QDs + HWR + IgY [ mg/mL]
The conditions of gel electrophoresis were optimized:
- running buffer Tris Tricine with pH adjusted to 9,
- loading buffer with mercaptoethanol (stabilizing agent)
Optimal ratios of HWR and antibody were found 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. Investice do rozvoje vzdělávání
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