1. NanoMed 2009, Berlin Nanoparticle-bioconjugates by laser ablation- a novel method aiming at pure drug and gene delivery S. Petersen, S. Barcikowski JRG Nanoparticles

2. NanoMed 2009, Berlin Nano Medicine Imaging Drug Delivery Therapy Targeting Gao et al. 2004 © MagForce Nanotechnolgies Nanobiomaterials Science Photogallery

3. NanoMed 2009, Berlin Imaging Drug Delivery Therapy Targeting Gao et al. 2004 © MagForce Nanotechnolgies Nanobiomaterials Science Photogallery Core/Shell Nanoparticle

4. NanoMed 2009, Berlin Functional Nanoparticles Core : Low toxicity, stable, visualizable, easily conjugable (e.g. gold nanoparticles) Coating: Tailored biological function Prerequisite for Nanomedicine Requirements for Production Method No side effects, cytotoxicity Rapid (single step) preparation method Size control Material variety Multifunctionality

5. NanoMed 2009, Berlin Femtosecond laser ablation in liquids 100% pure Material variety Fully dispersed and stable Safe fs-laser

6. NanoMed 2009, Berlin Electron affinity of laser generated gold nanoparticles fs-laser variable target (e.g. Au) Au +, Au 3+  Nanoparticle = electron acceptor Ligand = electron donor  Conjugation SH COOH NH 2 Compagnini, G et al, J. Mater. Res. 2004, Vol.19, No.10 Meunier group: Journal Am. Chem. Soc. 2004, Vol 126, S. 7176 – 7177 S. Petersen, J. Jakobi, S. Barcikowski. Biomaterialien 8 (3) 2007. 155-156 S. Petersen, S. Barcikowski. Appl. Surf. Sc. (2008). in press Mafune group, J. Phys. Chem. C, 2007, 10.1021 Meunier group, J. Phys. Chem. B, 2004, 108, 16864-16869

7. NanoMed 2009, Berlin In-situ bioconjugation e.g. gold nanoparticles with thiolized single stranded oligonucleotides (23mer) Au S S S S S S S S S fs laser beam variable target (e.g. Au) variable reagent (e.g. thiolized ssoligonucleotide)

8. NanoMed 2009, Berlin 1.) Negligible degradation of biomolecule Gel electrophoresis Deg (x) = 1- I(x)/ I(0) Evaluating optimal laser and process parameter 2.) Sufficient nanoparticle productivity SH

9. NanoMed 2009, Berlin Degrades the biomolecule during laser irradiation? SH Petersen et al. Adv. Funct. Mater. 2008

10. NanoMed 2009, Berlin Full integrity of biomolecules can be guaranteed Degrades the biomolecule during laser irradiation? SH Petersen et al. Adv. Funct. Mater. 2008

11. NanoMed 2009, Berlin Productivity during in-situ bioconjugation Productivity SH Petersen et al. APSUSC 2008

12. NanoMed 2009, Berlin Productivity and Purity during in-situ bioconjugation Productivity Integrity 100% integrity of DNA at moderate nanoparticle productivity: 1mg/h SH Petersen et al. APSUSC 2008

13. NanoMed 2009, Berlin Gold- DNA- Conjugate Gold Au Petersen et al. JLMN 2009 UV-Vis Spectra of Bioconjugates SH

14. NanoMed 2009, Berlin 200 nm 10 nm 200 nm Petersen et al. Adv. Funct. Mater. 2008 Size quenching leading to monodisperse bioconjugates SH

15. NanoMed 2009, Berlin Tailored Sizes of Bioconjugates SH Petersen et al. Adv. Funct. Mater. 2008

16. NanoMed 2009, Berlin Tailored Sizes of Bioconjugates Screening of different sizes in biomedical assays is enabled SH Petersen et al. Adv. Funct. Mater. 2008

17. NanoMed 2009, Berlin Petersen et al. APSUSC 2008 In-situ bioconjugation with unmodified oligonucleotides 100 nm  Monodisperse Bioconjugates

18. NanoMed 2009, Berlin Successful biological recognition of DNA (129bp) by PCR No alteration due to laser irradiation In-situ bioconjugation with double stranded DNA HPRT-DNA: canine Hypoxanthin- Guanin-Phosphoribosyltransferase *In collaboration with H. Murua Escobar, J. Soller Tierärztliche Hochschule Hannover, unpublished 100bp Negative control HPRT-DNA (129bp) AuNP-HPRT conjugates * SH

19. NanoMed 2009, Berlin In-situ Coating with Polymers and Silica 10 nm Silica FeO x PVP 8 mM TEOS, NH 3 (5 times) 100 nm Silica PVP FeO x S. Barcikowski, S. Petersen, A. Schnoor, unpublished

20. NanoMed 2009, Berlin In-situ Bioconjugation during laser ablation......enables the generation of pure bioconjugates...is a rapid (single step) preparation method...allows a size-tuning of bioconjugate...offers the possibility of screening various nanoparticle-bioconjugates...Multifunctionality ???

21. NanoMed 2009, Berlin In-situ bioconjugation - Summary Novel single step approach to pure nanoparticle-bioconjugates High material variety (core-material and biomolecule-shell) High purity Size control (via fluence) Rapid preparation method

22. NanoMed 2009, Berlin Thank you ! s.petersen@lzh.de Contributions from Group 'Nanomaterials' of LZH Stephan Barcikowski Niko Bärsch Annette Barchanski Anne Hahn Jurij Jakobi Ana Menendez Christin Menneking Laszlo Sajti Svea Petersen Ramin Sattari Andreas Schwenke Arne Schnoor Philipp Wagener Jürgen Walter Johanna Walter Financial support DFG-Excellence Cluster REBIRTH Junior Research Group 'Nanoparticles' Contributions from TIHO (I. Nolte) J. Soller H. Murua Escobar

23. NanoMed 2009, Berlin In organic Solvents, long pulses may cause pyrolysis ControlGC/MS Control Nd:YAG Laser, 800 ns t=30min, r=5kHz, f=100mm, V=3mL, E=1.5mJ GC/MS Control fs-Laser, 100 fs t=30min, r=5kHz, f=100mm, V= 3mL, E=300µJ fs pulses sub-µs/ns pulses

24. NanoMed 2009, Berlin Ultrashort laser pulses give pure structures Equilibrum temperature, heat conduction Ablation with nanosecond pulses 0 s10 -15 s10 -12 s10 -9 s10 -6 s 10 -3 s 10 ns Laser Pulse Duration Photons-Electrons Electrons-Atoms Heat Conduction 25 Nanosecond pulse laser ablated hole in silicon No thermal load, multiphoton effects Ablation with femtosecond pulses 0 s10 -15 s10 -12 s10 -9 s10 -6 s 10 -3 s 120 fs Laser Pulse Duration Electrons-Atoms (Lattice) Photons-Electrons 150 Femtosecond pulse laser ablated hole in silicon

25. NanoMed 2009, Berlin Selecting Appropriate Laser Ablation Methods for Nanoparticle Production Picosecond Laser Ablation Ablation of Bulk Material in Liquids and Gases, but: stability issues Femtosecond Laser Ablation (Stoichiometric) Ablation of Bulk Material (Metals, Metal Oxides, Alloys) in Liquids and gases IR-Laser Ablation (Continuous Wave) Generation of 50 – 500 nm Aerosols (e.g. Metals, Polymers) Quality and Versatility Productivity IR-Laser Ablation (ns pulsed) Generation of nanoarticles, but: thermal load of liquid and target

26. NanoMed 2009, Berlin Bioconjugation Au SH In situ functionalization of gold nanoparticles with thiol terminated ssoligonucleotides