MAGNETISMO E AFINIDADE PARA PROCESSOS BIOTECNOLGICOS MAGNETISM AND
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
MAGNETISM AND BIOMIMETISM IN BIOTECHNOLOGY NANOMATERIALS MAGNETISM BIOPOLYMERS AFFINITY BIOSEPARATION PROCESSES BIOCATALYSIS BIOMEDICAL APPLICATIONS Cecília Roque 6 th July 2010
MAGNETIC NANOPARTICLES Cecília Roque 6 th July 2010
SUPERPARAMAGNETISM Cecília Roque 6 th July 2010
MAGNETIC NANOMATERIALS (BIO)POLYMERIC SHELL FUNCTIONALITY MAGNETIC CORE – Fe 3 O 4 Cecília Roque 6 th July 2010
MAGNETIC NANOMATERIALS Groups that can be used to anchor polymers/molecules NPG Asia Mater. 2(1) 23– 30 (2010) Cecília Roque 6 th July 2010
BIOPOLYMERS BIOPOLYMERIC SHELL q Made of simple biological compounds q Produced by living organisms (natural and engineered sources) q Renewable materials, generally non-toxic and biodegradable q Excellent functional properties q Biocompatible SUPERPARAMAGNETIC CORE POLYSACCHARIDES Cecília Roque 6 th July 2010
BIOPOLYMERS – GUM ARABIC • Natural composite (exudate trees) • Highly branched polysaccharide & arabinogalactan-protein complex Gold Nanoparticles Carbon Nanotubes Quantum Dots Iron Oxide Magnetic Nanoparticles Cecília Roque Biocompatibility Stabilization 6 th July 2010
MNP SURFACE COATING Bare magnetite (MNP_GAadsorbed) GA co-precipitated MNPs (MNP_GA) Aldehyde-activated MNPs (MNP_GAAPTES) Diameter of individual core (11 -14 nm) 300 - 1000 nm Aminated MNP (MNP_GAEDC) Journal of Biotechnology, 144 (2009), pp. 313 -320 Cecília Roque 6 th July 2010
MAGNETIC NANOPARTICLES AND GA a) HEK 293 cells (control), b) HEK 293 cells + MNP, c) HEK 293 cells + MNP_GAADS Bar represents 50 µm Bicho et al, J. Mol. Recognit. , (in press) COATING WITH GA INFLUENCES CELLULAR GROWTH Cecília Roque 6 th July 2010
AFFINITY MAGNETIC FISHING NANOPARTICLES MAGNETISM AFFINITY NEW BIOSEPARATION PROCESSES MAGNETIC FISHING Cecília Roque 6 th July 2010
BIOSEPARATIONS 70% Downstream processing (Bioseparation) Others CANCER AUTO-IMMUNE DISEASES TREATMENT AND DIAGNOSTICS Cecília Roque HIGH COST BIOPHARMACEUTICALS 6 th July 2010
AFFINITY MAGNETIC FISHING contaminants Cecília Roque PRODUCT 6 th July 2010
AFFINITY MAGNETIC FISHING GUM ARABIC MAGNETIC CORE MAGNETIC NANOPARTICLES WITH AFFINITY LIGANDS Cecília Roque AFFINITY LIGANDS 6 th July 2010
AFFINITY LIGANDS BIOLOGICAL PRODUCT BIOLOGICAL RECEPTOR BIOMIMETISM SYNTHETIC RECEPTOR -STABILITY - LOW COST Cecília Roque 6 th July 2010
ANTIBODY MAGNETIC FISHING GUM ARABIC MAGNETIC CORE R 1 R 2 SYNTHETIC AFFINITY LIGANDS WITH AFFINITY FOR HUMAN Ig. G • LIGAND 22/8 • LIGAND 8/7 Batalha et al, J. Mol. Recognit. , (in press) Cecília Roque 6 th July 2010
ANTIBODY MAGNETIC FISHING Ka = 1. 5 x 105 M-1 Qmáx = 344 ± 21 mg Ig. G/g support Human Ig. G (mg/g) Batalha et al, J. Mol. Recognit. , (in press) Cecília Roque AGAROSE 0 MNP_GA 20 MNP 50 6 th July 2010
MAIN CONCLUSIONS q MNPs can be coated with GA by adsorption and covalent interactions q The diameter of the MNPs magnetic core was between 11 -14 nm. Hydrodynamic diameter ranges 300 -1000 nm q Reduced cytotoxicity of MNPs coated with GA q MNP_GA present low non-specific binding when compared to bare magnetite q Successful modification of the GA shell with biological ligands (protein-protein interaction) q MNP_GA successfully functionalized with two different triazine-based ligands (Ig. G) q Affinity to Ig. G comparable to agarose based systems q Higher amount of Ig. G bound per mass of support (compared agarose) Cecília Roque 6 th July 2010
ACKNOWLEDGEMENTS Olga Iranzo (ITQB, Portugal) António Lopes (ITQB, Portugal) Chris Lowe (Io. B, 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 B 37/08 REQUIMTE (DQ/FCT, Portugal) Cecília Roque 6 th July 2010
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