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Cecília Roque

MAGNETISMO E AFINIDADE PARA PROCESSOS BIOTECNOLÓGICOS MAGNETISM AND AFFINITY FOR BIOTECHNOLOGICAL PROCESSES. A. Cecília Roque. REQUIMTE - Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal. Cecília Roque. 6 th July 2010.

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Cecília Roque

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  1. MAGNETISMO E AFINIDADE PARA PROCESSOS BIOTECNOLÓGICOS MAGNETISM AND AFFINITY FOR BIOTECHNOLOGICAL PROCESSES A. Cecília Roque REQUIMTE- Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal Cecília Roque 6th July 2010

  2. MAGNETISM AND BIOMIMETISM IN BIOTECHNOLOGY Cecília Roque 6th July 2010

  3. MAGNETIC NANOPARTICLES Cecília Roque 6th July 2010

  4. SUPERPARAMAGNETISM Cecília Roque 6th July 2010

  5. MAGNETIC NANOMATERIALS (BIO)POLYMERIC SHELL FUNCTIONALITY MAGNETIC CORE – Fe3O4 Cecília Roque 6th July 2010

  6. MAGNETIC NANOMATERIALS Groups that can be used to anchor polymers/molecules NPG Asia Mater. 2(1) 23–30 (2010) Cecília Roque 6th July 2010

  7. BIOPOLYMERS BIOPOLYMERIC SHELL • Made of simple biological compounds • Produced by living organisms (natural andengineeredsources) • Renewable materials, generally non-toxic and biodegradable • Excellent functional properties • Biocompatible SUPERPARAMAGNETIC CORE POLYSACCHARIDES Cecília Roque 6th July 2010

  8. BIOPOLYMERS – GUM ARABIC • Natural composite (exudate trees) • Highly branched polysaccharide & arabinogalactan-protein complex Gold Nanoparticles Carbon Nanotubes Quantum Dots Iron Oxide Magnetic Nanoparticles Biocompatibility Stabilization Cecília Roque 6th July 2010

  9. MNP SURFACE COATING Bare magnetite (MNP_GAadsorbed) GA co-precipitated MNPs (MNP_GA) Diameter of individual core (11-14 nm) Aldehyde-activated MNPs (MNP_GAAPTES) 300 - 1000 nm Aminated MNP (MNP_GAEDC) Journal of Biotechnology, 144 (2009), pp. 313-320 Cecília Roque 6th July 2010

  10. MAGNETIC NANOPARTICLES AND GA a) HEK293 cells (control), b) HEK293 cells + MNP, c) HEK293 cells + MNP_GAADS Bar represents 50 µm Bicho et al, J. Mol. Recognit., (in press) COATING WITH GA INFLUENCES CELLULAR GROWTH Cecília Roque 6th July 2010

  11. AFFINITY MAGNETIC FISHING NEW BIOSEPARATION PROCESSES MAGNETIC FISHING Cecília Roque 6th July 2010

  12. BIOSEPARATIONS 70% TREATMENT AND DIAGNOSTICS HIGH COST BIOPHARMACEUTICALS CANCER AUTO-IMMUNE DISEASES Cecília Roque 6th July 2010

  13. AFFINITY MAGNETIC FISHING contaminants PRODUCT Cecília Roque 6th July 2010

  14. AFFINITY MAGNETIC FISHING GUM ARABIC MAGNETIC CORE AFFINITY LIGANDS MAGNETIC NANOPARTICLES WITH AFFINITY LIGANDS Cecília Roque 6th July 2010

  15. AFFINITY LIGANDS BIOLOGICAL PRODUCT BIOLOGICAL RECEPTOR • BIOMIMETISM • SYNTHETIC RECEPTOR • STABILITY • LOW COST Cecília Roque 6th July 2010

  16. ANTIBODY MAGNETIC FISHING GUM ARABIC • SYNTHETIC AFFINITY LIGANDS WITH AFFINITY FOR HUMAN IgG • LIGAND 22/8 • LIGAND 8/7 MAGNETIC CORE R1 R2 Batalha et al, J. Mol. Recognit., (in press) Cecília Roque 6th July 2010

  17. ANTIBODY MAGNETIC FISHING Ka = 1.5x105 M-1 Qmáx = 344 ± 21 mg IgG/g support Batalha et al, J. Mol. Recognit., (in press) Cecília Roque 6th July 2010

  18. MAIN CONCLUSIONS • MNPs can be coated with GA by adsorption and covalent interactions • The diameter of the MNPs magnetic core was between 11-14 nm. Hydrodynamic diameter ranges 300-1000 nm • Reduced cytotoxicity of MNPs coated with GA • MNP_GA present low non-specific binding when compared to bare magnetite • Successful modification of the GA shell with biological ligands (protein-protein interaction) • MNP_GA successfully functionalized with two different triazine-based ligands (IgG) • Affinity to IgG comparable to agarose based systems • Higher amount of IgG bound per mass of support (compared agarose) Cecília Roque 6th July 2010

  19. ACKNOWLEDGEMENTS Olga Iranzo (ITQB, Portugal) António Lopes (ITQB, Portugal) Chris Lowe (IoB, Univ. Cambridge, UK) Raquel Aires-Barros (IBB, IST, Portugal) M. Ascensão Reis, R. Oliveira (REQUIMTE, PT) Pedro Simões, P. Vidinha (REQUIMTE; PT) Dora Pereira (MRC, HNR, UK) Fundação para a Ciência e Tecnologia (FCT) PTDC/BIO/65383/2006 PTDC/EBB-BIO/102163/2008 PTDC/EBB-BIO/098961/2008 SFRH/BPD/34768/2007 SFRH/BD/48804/2008 Banco Santander Totta and UNL British Council/CRUP, through Acção Integrada B37/08 REQUIMTE (DQ/FCT, Portugal) Cecília Roque 6th July 2010

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