磁珠在临床以及生物科学与技术中的运用.ppt

Magnetic nano-and microparticles in bioscience and biotechnology,Type of magnetic nano-and microparticles,Multi domain,single domain or superparamagneticMagnetite(Fe3O4)Ferrites(MeO.Fe2O3;Me=Ni,Co,Mg,Zn,Mn)Maghemite(-Fe2O3)Greigite(Fe3S4)Iron,nickel,Magnetic nano-and microparticles,Inorganic material,but:Present in biological systemsUsable for a variety of bio-applications,Important dates,1962-Lowenstam(biochemically-precipitated magnetite as a capping material in the radula(tongue plate)teeth of chitons(marine mollusks of the class Polyplacophora)1975-Blakemore(magnetotactic bacteria),Magnetic material in teeth of chitons,hardening the tooth caps(enabling the chitons to extract and eat endolithic algae from within the outer few millimeters of rock substrates),Magnetotactic bacteria,Various morphological types existGram-negative prokaryotesMagnetic nanoparticles present in magnetosomesFound in sediments in diverse aquatic environments,Magnetotactic bacteria,Provided by Dr.D.Schler,Germany,Magnetosomes,Composed from magnetite(Fe3O4)or greigite(Fe3S4)Ca 50 nm in diameterCovered with a lipidic membraneImportant for navigation of the bacteria into oxic/anoxic layer Biocompatible character(phospholipid bilayer)Possible applications in biosciences,Magnetosomes,Provided by Dr.D.Schler,Germany,Magnetosomes,Provided by Dr.D.Schler,Germany,Biogenic magnetic nanoparticles,Magnetotactic bacteriaMagnetotactic algae ProtozoaInsectFishAmphibiansReptilesBirdsMammals,Animal magnetite,Magnetosome chains from the frontal tissues of chinook salmon S.Mann et al.:J.Exp.Biol.140(1988)35-49.,Importance of animal magnetite,Most probably involved in magnetoreception(what is the current position?,which direction to move?)interaction of magnetite nanoparticles(chains of magnetic nanoparticles?)with the sensing cells,Non-physiological presence of magnetic nanoparticles,Found in human brain in case of neurodegenerative diseasesDisruption of normal iron homeostasis in the brainEarly detection of neurodegenerative diseases using MRI?,Magnetic nanoparticles in meteorites from Mars,Meteorite magnetic nanoparticles resemble magnetosomes from Earth magnetotactic bacteriaHypothesis:Was there a transfer of living systems from Mars to Earth?,Lets return back to Earth!,Magnetic nano-and microstructured materials for bioapplications,Useful for many applications in biosciences,biotechnologies and environmental technologiesEnable to label diamagnetic structures Enable separation or detection of magnetically labeled structuresBoth separation and non-separation applications are important,Which nano-and microstructured magnetic materials?,Three important bio-Biocompatible(in the broadest sense of this word)BiodegradableBiomimetic,Biocompatible materials,Biocompatibility in the broadest sense of the wordMinimal toxicity Stability of nanoparticles in body fluidsPossibility for immobilization of biologically active compounds,cell organelles,cells(activated nanoparticles)Reproducible interaction with biosystemsEnable labeling of biostructures(for magnetoassays)Biopolymer based particles,PEGylated structures,magnetoliposomes,biopolymer-based FFs,self-assembled magnetic structures,Biodegradable materials,Important from the point of view of possible clinical applications(e.g.,drug targeting)Nanoparticles formed by biodegradable(bio)polymerMagnetoliposomes and related structures Starch-based magnetic fluids,Biomimetic materials,Analogues of bacterial magnetosomesFerritin x magnetoferritin,Important data,1960s Pappell(first ferrofluids,1000 hours of magnetite milling)1966-Magnetically controlled thrombosis of intracranial aneurysms1970s increase of interest in biomagnetic research and technology1980 Massart(chemical synthesis of aqueous ferrofluid without detergents)1980s commercially available magnetic particles,Magnetic fluids examples of magnetic nanostructured materials,ferrofluid can be influenced by the presence of external magnetic field,Synthetic biocompatible magnetic nanoparticles and complexes containing them,Biocompatible magnetic fluidsMagnetoliposomesMagnetic nano-and microparticlesMagnetoferritin,Magnetic microparticles,DynabeadsHomogeneous size distribution Simple recovery with small permanent magnetsMagnetic nanoparticles present inside the beadsAvailable in plain,activated and bioactive form,Examples of of magnetic nano-and microparticles applications,From molecular biology to environmental technologies Manipulation of microliters as well as million of liters Manipulation in suspension systemsBoth separation and non-separation techniques are important,Immobilization and modification of biologically active compounds,Possibility of simple removal of the immobilized molecules or cells from the system Immobilization of enzymes,antibodies,lectins,oligonucleotides,affinity ligandsModification of proteins via magnetic PEGImmobilization in magnetoliposomesMagnetosomes from genetically engineered magnetotactic bacteria carrying target proteins,Isolation of biologically active compounds,Direct separation of targets from crude samples containing solidsAffinity magnetic beads or magnetic two-phase systemsPreconcentration of target molecules from large volumes of crude materials Poly A+mRNA,DNA,oligonucleotides EnzymesLectinsDNA/RNA binding proteinsAntibodies,antigens Xenobiotics(both organic and inorganic),High gradient magnetic separators,Test tube magnetic separators,Automated systems,KingFisher mLThermo,USA,KingFisher 96Thermo,USA,Te-MagSTecan,Japan,Determination and detection of biologically active compounds and xenobiotics,Immunomagnetic assays(magnetic particles used for separation of target analyte)Magnetoimmunoassays(magnetic particles serve as detectable labels)Magnetic substratesMagnetic solid-phase extractionNew principles for the detection and quantitation of both magnetic labels and biological structures tested(giant magnetoresistance,magnetoelasticity,magnetic atomic force microscopy),Automated systems for immunoassays,Combination of specific magnetic separation and sensitive detection(enzyme,radioactive,chemiluminescent or fluorescent)Different assay formats can be used(sandwich x competitive immunoassays)Generic solid phases can be used(streptavidin,secondary antibody,protein A),Beckman Coulters Access Immunoassay System,Magnetoimmunoassays,Magnetic nanoparticles serve as a labelPossibility to replace enzymes,radioisotopes Magnetic Permeability Reader(Euris,Sweden,top;Magnetic Assay Reader(Quantum design,USA,bottom),Magnetic solid-phase extraction,Detection of crystal violet in water Magnetite with immobilized phthalocyanine dye used as a selective adsorbentCurve B 0.5 g of dye in 1000 ml of water Sensitivity 0.5 ppb achieved,Safarik,I.,Safarikova,M.:Detection of low concentrations of malachite green and crystal violet in water.Water Res.36(2002)196,Cells,Immunomagnetic separation of target cells and cell organelles Detection and separation of cancer cellsIsolation of stem cellsSeparation of microbial pathogens Study of cell function Transfection of cells Magnetic twisting cytometry Magnetic labeling of cellsMRI tracking of transplanted cells(magnet.modified),Labeling of T-lymphocytes with magnetic nano-and microparticles,E.coli O157 bound to Dynabeads,Principle of IMS,Large-scale immunomagnetic separation of cells,Several magnetic separation devices commercially available CliniMACS employs magnetic nanoparticles for cells labeling and HGMS system for separation,Magnetotactic bacteria,Biotechnology production of native or genetically engineered magnetosomesNon-destructive domain analysisRadionuclide recovery,Drug and radionuclide targeting,Accumulation of active compounds in the target organ or tumor Ferrofluids,magnetoliposomes,ferrofluid modified erythrocytes,magnetic polymer composites,Magnetic drug targeting,MTC-composite of elemental iron and activated carbon with anticancer drug adsorbed onto it(FeRx Inc.,San Diego,CA,USA),Magnetic Field,Body surface,Magnetic fluid hyperthermia,Cancer treatmentHeating of tissues labeled with magnetic particles to 42-46 C after exposure to alternating magnetic fieldMagnetic fluids,magnetoliposomes,magnetic single-or multi-domain particlesCombination with drug targeting,Sketch of the first prototype MFH therapy system(MFH Hyperthermiesysteme GmbH,Berlin,Germany)Prof.Jordan,Contrast increasing during MRI,Superparamagnetic contrast agentsBiocompatible ferrofluids,magnetoliposomesAntibody-conjugated magnetite nanoparticles,MRI of bone marrow,PEGylated magnetoliposomes as a bone marrow-seeking MR contrast agent,Potential medical applications,Treatment of AIDSEndocytosis of ferrofluids by cancer cellsBrain tumor sensingArtificial sphincter muscleMagnetic polymers(artificial muscle?),Environmental chemistry and technology,Waste water and potable water purification using magnetite microparticles(Sirofloc process Australia)Magnetically modified microbial cells for xenobiotics removalMagnetic solid-phase extraction for the preconcentration of the target analytesMagnetically modified enzymes for the detection of their inhibitors(heavy metals,pesticides etc),Scheme of Sirofloc Process,Inhibition of magnetically modified trypsin,Dependence of the inhibition of the trypsin activity on the concentration of Ag+()and Pb2+().Dependence of the inhibition of the trypsin activity on the concentration of thionin(),4-aminobenzamidin(),safranin()and bacitracin().,Adsorption of magnetic nanoparticles on yeast cells,Interaction of Saccharomyces cerevisiae cells(bakers yeast,brewers yeast)with water-based ferrofluidsMagnetically modified yeast cells can be used as an adsorbent for the separation of biologically active compounds and xenobiotics(both organic and inorganic)Adsorption of organic dyes is presented,Future trends,Biotechnology production of magnetic non-modified and modified nanoparticlesMedical applicationsMagnetic nanoparticles as labelsMagnetic nanotubes Biotechnology Integration of magnetic nano/microparticles with other progressive technologies(microfluidics,chips,MEMS.),Biomagnetic research and technology,Multidisciplinary research area leading to new resultsBoth basic research and important applications Creativity and fantasy is needed Biomagnetic research and technology is an emerging scientific discipline!,Review papers,Safarik,I.,Safarikova,M.:Magnetic nanoparticles and biosciences.Monatsh.Chem.133(2002)737-759Safarikova,M.,Safarik,I.:The application of magnetic techniques in biosciences.Magn.Electr.Sep.10(2001)223-252 Safarik,I.,Safarikova,M.:Use of magnetic techniques for the isolation of cells.J.Chromatogr.B 722(1999)33-53 Available on www page:http:/www.uek.cas.cz/people/safarikThe WWW page includes also other biomagnetic information,A new on-line journal,Journal WWW page:Published by BioMed Central()Edited by Ivo Safarik and Mirka Safarikova,Thank you for your attention!,