WO1991009678A1 - Particules revetues d'un polymere organo-metallique utilisees dans des separations - Google Patents

Particules revetues d'un polymere organo-metallique utilisees dans des separations Download PDF

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WO1991009678A1
WO1991009678A1 PCT/US1990/007492 US9007492W WO9109678A1 WO 1991009678 A1 WO1991009678 A1 WO 1991009678A1 US 9007492 W US9007492 W US 9007492W WO 9109678 A1 WO9109678 A1 WO 9109678A1
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particle
organo
titanium
iron
magnetically responsive
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PCT/US1990/007492
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English (en)
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Mark S. Chagnon
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Omni Quest Corporation
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
    • A61K49/1821—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
    • A61K49/1824—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
    • A61K49/1827—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
    • A61K49/183—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with an inorganic material or being composed of an inorganic material entrapping the MRI-active nucleus, e.g. silica core doped with a MRI-active nucleus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/06—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations
    • A61K49/18—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes
    • A61K49/1818—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles
    • A61K49/1821—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles
    • A61K49/1824—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles
    • A61K49/1827—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle
    • A61K49/1833—Nuclear magnetic resonance [NMR] contrast preparations; Magnetic resonance imaging [MRI] contrast preparations characterised by a special physical form, e.g. emulsions, microcapsules, liposomes particles, e.g. uncoated or non-functionalised microparticles or nanoparticles coated or functionalised microparticles or nanoparticles coated or functionalised nanoparticles having a (super)(para)magnetic core, being a solid MRI-active material, e.g. magnetite, or composed of a plurality of MRI-active, organic agents, e.g. Gd-chelates, or nuclei, e.g. Eu3+, encapsulated or entrapped in the core of the coated or functionalised nanoparticle having a (super)(para)magnetic core coated or functionalised with a small organic molecule
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B03—SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATIC TABLES OR JIGS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C—MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLID MATERIALS OR FLUIDS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS
    • B03C1/00—Magnetic separation
    • B03C1/005—Pretreatment specially adapted for magnetic separation
    • B03C1/01—Pretreatment specially adapted for magnetic separation by addition of magnetic adjuvants
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
    • G01N33/54326—Magnetic particles
    • G01N33/5434—Magnetic particles using magnetic particle immunoreagent carriers which constitute new materials per se
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/551—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2446/00—Magnetic particle immunoreagent carriers
    • G01N2446/20—Magnetic particle immunoreagent carriers the magnetic material being present in the particle core
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2446/00—Magnetic particle immunoreagent carriers
    • G01N2446/80—Magnetic particle immunoreagent carriers characterised by the agent used to coat the magnetic particles, e.g. lipids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2446/00—Magnetic particle immunoreagent carriers
    • G01N2446/80—Magnetic particle immunoreagent carriers characterised by the agent used to coat the magnetic particles, e.g. lipids
    • G01N2446/84—Polymer coating, e.g. gelatin
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2446/00—Magnetic particle immunoreagent carriers
    • G01N2446/80—Magnetic particle immunoreagent carriers characterised by the agent used to coat the magnetic particles, e.g. lipids
    • G01N2446/90—Magnetic particle immunoreagent carriers characterised by the agent used to coat the magnetic particles, e.g. lipids characterised by small molecule linker used to couple immunoreagents to magnetic particles

Definitions

  • magnetic particles When used as solid supports in immunoassays or other competitive binding assays, magnetic particles permit the reaction to occur under homogeneous conditions, which promotes optimal binding kinetics, minimally alters analyte-adsorbent equilibrium and facilitates separation of bound from unbound analyte, particularly as compared to centrifugation.
  • Centrifugal separations are time-consuming, require expensive and energy-consuming equipment and pose radiological, biological and physical hazards. Magnetic separations are relatively rapid and easy, requiring simple equipment.
  • the use of non-porous adsorbent- coupled magnetic particles in affinity chromatography systems allows better mass transfer and results in less fouling of the sample than in conventional affinity chromatography systems.
  • Small magnetic particles e.g., having a mean diameter in solution less than about 0.03 microns
  • the magnetic field and magnetic field gradient required to remove such particles from solution are large and require heavy and bulky magnets for generating these fields, which are inconvenient to use in bench top work.
  • Magnets capable of generating magnetic fields in excess of 5000 Oersteds, for example, are typically required to separate magnetic particles of less than 0.03 microns in diameter.
  • the ferromagnetic carrier particles must generally be coated in order to provide a reactive substrate for attaching chemical functional groups to bind enzymes or antibodies, for example.
  • Silane polymers are often used for this purpose.
  • Particles described in U.S. Patent No. 3,933,997 are coated with silanes capable of reacting with anti-digoxin antibodies to chemically couple the antibodies to the carrier particles.
  • Various silane couplings are discussed in U.S. Patent No. 3,652,761, which is hereby incorporated by reference. Procedures for silanization known in the art generally differ from each other in the media chosen for the polymerization of silane and its disposition on reactive surfaces.
  • the medium is generally an organic solvent such as toluene (H.W., Weetall, In: Methods of Enzymology, K. Mosbach (ed), 44:134-148, 140 (1976)), methanol (U.S. Patent No. 3,933,997) or chloroform (U.S. Patent No. 3,652,761). Silane depositions from aqueous alcohol and aqueous solutions with acid have also been used. H.W. Weetall, In: Methods in Enzymology, supra, p.139 (1976).
  • silane-coated particles There are several drawbacks to silane-coated particles.
  • the dehydration methods used to dry the coatings such as air and/or oven drying, allow the silanized surfaces of the carrier particles to contact each other, potentially resulting in interparticle bonding, including cross-linking between particles by siloxane formation, van der Waals interactions or physical adhesion between adjacent particles.
  • This interparticle bonding yields covalently or physically bonded aggregates of silanized carrier particles of considerably larger diameter than individual carrier particles.
  • Such aggregates have low surface area per unit weight and hence, a low capacity for coupling with molecules such as antibodies, antigens or enzymes. Such aggregates also have gravitational settling times which are too short for many applications.
  • Radioimmunoassay is a term used to describe methods for analyzing the concentrations of substances involving a radioactively labeled substance which binds to an antibody. The amount of radioactivity bound is altered by the presence of an unlabeled test substance capable of binding to the same antibody. The unlabeled substance, if present, competes for binding sites with the labeled substance and thus decreases the amount of radioactivity bound to the antibody. The decrease in bound radioactivity can be correlated to the concentration of the unlabeled test substance by means of a standard curve.
  • An essential step of RIA is the separation of bound and free label which must be accomplished in order to quantitative the bound fraction.
  • Coated tubes such as described in U.S. Patent No, 3.646,346 allow separation of bound and free label without centrifugation but suffer from two major disadvantages.
  • Antibodies have been attached to particulate systems to facilitate separations.
  • U.S. Patent Nos. 3,652,761 and 3,555,143 Such systems have large surface areas permitting nearly unlimited amounts of antibody to be used, but the particulates frequently settle during the assay. The tube frequently must be agitated to achieve even partial homogeneity P.M. Jacobs, The Ligand Quarterly, 4(4): 23-33 (1981). Centrifugation is still required to effect complete separation of bound and free label.
  • Antibodies may react with labeled and unlabeled molecules followed by separation using a second antibody raised to the first antibody.
  • the technique termed the doubled antibody method, achieves homogeneity of antibody during reaction with label but requires an incubation period for reaction of first and second antibodies followed by a centrifugation to pellet the antibodies.
  • Antibodies have been attached to magnetic supports in an effort to eliminate the centrifugation steps in radioimmunoassays for nortriptyline, methotrexate, digoxin, thyroxine and human placental lactogen.
  • Antiestradiol antibodies were coupled to such particles and their potential usefulness in estradiol RIAs was demonstrated. While this approach may have over come the problem of settling, the particle size and the magnetic coating nonetheless present limitations on surface area and hence limitations on the availability of sites for antibody coupling.
  • Magnetic separations have been applied in other biological systems besides RIA.
  • Several nonisotopic immunoassays such as fluoroimmunoassays (FIA) and enzyme-immunoassays (EIA) have been developed which employ antibody-coupled (or antigen coupled) magne t i c particles.
  • FIA fluoroimmunoassays
  • EIA enzyme-immunoassays
  • the principle of competitive binding is the same in FIA and EIA as in RIA except that fluorophores and enzymes, respectively, are submitted for radioiso- topes as label.
  • Kamel et al. developed magnetizable solid-phase FIA's for cortisol and phenyltoin, respectively, utilizing ferromagnetic cellulose/iron oxide particles to which antibodies were coupled by cyanogen bromide activation.
  • a magnetizable solid phase non-immune radioassay for vitamin B12 has been reported by D.S. Ithakissios and D.O. Kubiatowicz Clin. Chem., 23(11) : 2072 - 2079 (1977).
  • the principle of competitive binding in non-immune radioassays is the same as in RIA with both assays employing radioiso topic labels.
  • RIA is based on the binding or interaction of certain biomolecules like vitamin B12 with specific or non-specific binding, carrier, or receptor proteins.
  • the magnetic particles of Ithakissios and Kubiatowicz were composed of barium ferrite particles embedded in a water-insoluble protein matrix.
  • magnetic particles have been used for a variety of other biological purposes.
  • magnetic particles have been used in cell sorting systems to isolate select viruses, bacteria and other cells from mixed populations.
  • Magnetic particles have also been used as the solid phase support immobilized enzyme systems.
  • Enzymes coupled to magnetic particles are contacted with substrates for a time sufficient to catalyze the biochemical reaction. Thereafter, the enzyme can be magne- tically separated from products and unreacted substrate and potentially can be reused.
  • Magnetic particles have been used as supports for ⁇ -chymotrypsin, ⁇ -galactos idase (U.S. Patent No. 4,152,210), hereby incorporated by reference) and glucose isomerase (U.S. Patent No. 4,343,901, hereby incorporated by reference) in immobilized enzyme systems.
  • the present invention relates to magnetically responsive particles coated with an organo-metallic polymer capable of binding bioaffinity adsorbents, and to their use in the separation of biological molecules from, or directed movement of the molecules in, the surrounding medium.
  • the organo-metallic coating is adsorbed onto or covalently bound to the magnetic particle.
  • the magnetic particles comprise a magnetically responsive metal, metal alloy, or metal oxide core surrounded by an organo-metallic polymer coating which is adsorbed or covalently bound to the particle.
  • the organo-metallic polymer is formed from an organo-metallic monomer, which is applied to the metal particle, and thermally cross-linked in situ to form an adsorbed or a covalently bound polymer coating.
  • Organo-titanium polymers are preferred, however, organo-metallic polymers formed from coordinate complexes of other transition metals, such as zirconium (Zr), hafnium (Hf), vanadium (V), tantalum (Ta) and niobium (Nb), or post-transition metals, such as tin (Sn) and antimony (Sb), can be used.
  • Zr zirconium
  • Hf hafnium
  • V vanadium
  • Ta tantalum
  • Nb niobium
  • post-transition metals such as tin (Sn) and antimony (Sb)
  • a wide variety of bioaffinity adsorbents can be covalently bonded to the organo-metallic polymer coating through selected coupling chemistries.
  • the invention relates to methods for the preparation of magnetically responsive particles comprising a metal, metal alloy or metal oxide core and an organo-metallic coating having an aliphatic moiety and an organic functionality to which a variety of organic and/or biological molecules can be coupled.
  • the particles, coupled or uncoupled, can be dispersed in aqueous media forming a colloidal dispersion which is stable, that is, the particles resist rapid gravitational settling.
  • the particles can be reclaimed from the media by applying a magnetic field.
  • the particles are superparamagnetic; that is, they exhibit no reminent magnetization after removal of a magnetic field which allows the particles to be redispersed without magnetic aggregate formation.
  • the organo-metallic coated magnetically responsive particles of the invention may be coupled through the organic functionality to biological or organic molecules with affinity for, or the ability to adsorb, or which interact with, certain other biological or organic molecules. Particles so coupled may be used in a variety of in vitro or in vivo systems involving s ep arat i ons steps or the directed movement of coupled molecules to particular sites, including immunological assays, other biological assays, biochemical or enzymatic reactions, affinity chromatographic purification, cell sorting and diagnostic and therapeutic uses.
  • the present organo-metallic coated magnetic particles provide superior composition, size, surface area, coupling versatility, settling properties and magnetic behavior for use in biological separations.
  • the magnetic particles of this invention are suitable for many of the assays, enzyme immobilization, cell sorting and affinity chromatography procedures reported in the literature and, in fact, overcome many of the problems associated with particle settling and reuse experienced in the past with such procedures.
  • the magnetically responsive particles of this invention overcome problems associated with the size, surface area, gravitational settling rate and magnetic character of previously developed magnetic particles. Gravitational settling times in excess of about 24 hours can be achieved with the present magnetic particles.
  • the gravitational settling time is defined to be the time for the turbidity of a dispersion of particles to fall by fifty percent in the absence of a magnetic field gradient.
  • the present magnetic particles comprise a core of a magnetically responsive metal, metal alloy or metal oxide, coated with organo-metallic polymer, which is capable of binding reactive groups or agents, for example, chemically reactive groups, biologically reactive groups or bioaffinity agents.
  • the organo- metallic polymer is adsorbed onto or covalently bound to the magnetic particle.
  • the term "magnetically responsive particle” or “magnetic particle” is defined as any particle dispersible or suspendible in aqueous media without significant gravitational settling, and separable from suspension by application of a magnetic field.
  • magnetic core is defined as a crystal or group (or cluster) of crystals of a transition metal, alloy or magnetic metal oxide having ferrospinel structure and comprising trivalent and divalent cations of the same or different transition metals or magnetic metal crystal group.
  • Metals, alloys and oxides which are useful as magnetic core material in the present invention include the metals, alloys and oxides based on metals which appear in the Periodic Table in Groups 4a and b, 5a and b, 6a and 7a.
  • divalent transition metals such as iron, magnesium, manganese, cobalt, nickel, zinc and copper
  • alloys of these metals such as iron alloys or oxides (e.g., iron magnesium oxide, iron manganese oxide, iron cobalt oxide, iron nickel oxide, iron zinc oxide and iron copper oxide), cobalt ferrite, samarium cobalt, barium ferrite, and aluminum - ni ckel - c ob al t and metal oxi de s inc luding magnetite (Fe 3 O 4 ), hematite (Fe 2 O 3 ) and chromium dioxide (CrO 2 ).
  • divalent transition metals such as iron, magnesium, manganese, cobalt, nickel, zinc and copper
  • alloys of these metals such as iron alloys or oxides (e.g., iron magnesium oxide, iron manganese oxide, iron cobalt oxide, iron nickel oxide, iron zinc oxide and iron copper oxide), cobalt ferrite, samarium
  • a magnetic core may be comprised of a cluster of superparamagnetic crystals of iron oxide, or a cluster of superparamagnetic or ferromagnetic crystals of irons or oxide, or may consist of an single superparamagnetic or ferromagnetic crystal of an iron oxide or metal alloy.
  • the present particles are preferably between about 0.003 and about 1.5 microns in diameter, and have a surface area of from about 50 to 150 meters/gm, which provides a high capacity for coupling of a bioaffinity adsorbent, chemical or biochemical reactive group.
  • Magnetic particles of this size range overcome the rapid settling problems of larger particles, but obviate the need for large magnets to generate the magnetic fields and magnetic field gradients required to separate smaller particles.
  • magnets used to effect separations of the magnetic particles of this invention need only generate magnetic fields between about 100 and about 1000 Oersteds. Such fields can be obtained with permanent magnets which are smaller than the container which holds the dispersion of magnetic particles and, thus, are suitable for benchtop use.
  • superparamagnetic particles do not exhibit the magnetic aggregation associated with ferromagnetic particles and permit redispersion and reuse.
  • superparamagnetism is defined as that magnetic behavior exhibited by iron, cobalt, nickel or other metal alloys or metal oxides having a crystal size of less than about 300A, which behavior is characterized by responsiveness to a magnetic field without reminant magnetization.
  • Ferromagnetic particles may be useful in certain applications of the Invention.
  • the term "ferromagnetism” is defined as that magnetic behavior exhibited by Iron, iron alloys or iron oxides with a crystal size greater than about 500A, which behavior is characterized by responsiveness to a magnetic field with a reminant magnetization of greater than about 10 gauss upon removal of the magnetic field.
  • Organo-metallic monomers useful for the present coated particles are organic coordinate complexes of selected transition and/or post transition metals which are capable of forming a stable coordination coumpound, and organic ligands, which can be adsorbed onto or covalently bound to the magnetic particle and, crosslinked in situ on the particle surface, thereby forming the organo-metallic polymer coating.
  • the organo-metallic monomer must be able to be functionalized or derivatized in a manner that allows the polymer formed therefrom to form covalent bonds with bioaffinity or chemical affinity adsorbents.
  • the organo-metallic polymer is post- functionalized or derlvitized with an aliphatic "spacer arm" which is terminated with an organic functional group capable of coupling with bioaffinity adsorbents.
  • the "spacer arm” is an aliphatic hydrocarbon having from about 2 to about 60 atoms, e.g., carbon, nitrogen and/or oxygen atoms.
  • the purpose of the spacer arm is to provide a non-reactive linker (or spacer) between the organic group which reacts with the chemical group, biochemical group or bioaffinity adsorbent and the polymer chain, and to impart an appropriate degree of hydrophilic/hydrophobic balance to the surface of the coated particle.
  • organo-metallic compounds are coordinate complexes formed from selected transition metals (e.g., Ti, Zr, Hf, V, Zn, Cd, Mn, Te, Re, Ta, Nb) and/or post-transition metals (e.g., Sn, Sb, Al, Ga, In, Ge).
  • transition metals e.g., Ti, Zr, Hf, V, Zn, Cd, Mn, Te, Re, Ta, Nb
  • post-transition metals e.g., Sn, Sb, Al, Ga, In, Ge.
  • Organo-titanium compounds are parti cul ar ly preferred.
  • Organo-titanium compounds which are us e ful including, for example, titanium - tetra - isopropoxide, amino - hexyl - titanium - tri - isopropoxide, amino-propyl- titanium - tri - isopropoxide and carboxyl -hexyl - titanium- tri - isopropoxide.
  • amino -hexyl - titanium- tri - isoproxide is coated onto the magnetic particle of choice, and thermally crosslinked to form an organo-titanium polymer coating having an aliphatic spacer arm (the hexyl moiety) and organic functional group (the amine group).
  • the coated particle is post-functionalized, if necessary, in a manner that allows the organo-metallic polymer to form covalent bonds with bioaffinity or chemical affinity adsorbents.
  • an organo-titanium polymer such as titanium- tetra- isopropoxide which lacks the spacer arm and organic functional group, is coated onto the magnetic particle of choice and partly crosslinked at about 40°C for a period of time sufficient to allow the organotitanium polymer to become adsorbed onto the particle surface.
  • the organotitanium coated magnetic particle is then activated by reaction with an agent such as 1-hydroxy- 6-amino hexane, to form the amino-hexyl- titanium- tri- isopropoxide.
  • the coating is then crosslinked at elevated temperatures to form an organotitanium polymer coating having an aliphatic spacer arm and an organic functionality (i.e., the amine group).
  • the functionalized particle can then be reacted or coupled, with the bioaffinity adsorbent of choice.
  • the magnetic core particles are prepared according to the following general procedure: metal salts are precipitated in a base to form fine magnetic metal oxide crystals. The crystals are redlspersed, then washed in water and in an electrolyte. Magnetic separation can be used to collect the crystals between washes if the crystals are superparamagnetic.
  • superparamagnetic iron oxide particles are made by precipitation of divalent (Fe 2+ ) and trivalent (Fe 3+ ) iron salts, for example, ferrous ammonium sulfate, Fe 2 (NH 2 )(SO 4 ) and ferric sulfate, Fe 2 (SO 4 ) 3 , in aqueous base.
  • the ratio of Fe 2+ and Fe 3+ and counterion can be varied wi thout substantial changes in the final product by increasing the amount of Fe 2+ while maintaining a constant molar amount of iron.
  • Counterions including nitrate, sulfate, chloride or hydroxide are useful in the method.
  • a Fe 2+ /Fe 3+ ratio of about 2:1 to about 4:1 is useful in the present invention; a ratio of about 2:1 Fe 2+ : Fe 3+ is particularly useful.
  • An Fe 2+ /Fe 3+ ratio of 1:1 produces magnetic particles of slightly inferior quality to those resulting from the higher Fe 2+ /Fe 3+ ratios, the particle size is more heterogeneous than that resulting from Fe 3+ /Fe 2+ of 2:1 or 4:1.
  • aqueous solutions of the iron salts are mixed in a base, such as ammonium, sodium or potassium hydroxide, which results in the formation of a crystalline precipitate of superparamagnetic iron oxide.
  • a base such as ammonium, sodium or potassium hydroxide
  • the precipitate is washed repeatedly with water by magnetically separating and redispersing it until a neutral pH is reached.
  • the precipitate is then washed with about five equal portions of a water miscible solvent, such as acetone, methanol or ethanol that has been dried over molecular sieves to remove all of the water.
  • divalent transition metal salts such as magnesium, manganese, cobalt, nickel, zinc and copper salts may be substituted for iron salts in the precipltation or milling procedure to yield magnetic metals or metal oxides.
  • divalent cobalt chloride (CoCl 2 ) for FeCl 2 in the above procedure produced ferromagnetic metal oxide particles. Ferromagnetic metal oxide particles such as those produced with CoCl 2 can be washed in the absence of magnetic fields by employing conventional techniques of centrifugation or filtration between washings to avoid magnetizing the particles.
  • the resulting ferromagnetic metal oxides are of sufficiently small diameter to remain dispersed in aqueous media, they can also be coated with the organo-metallic polymer and coupled to bioaffinity adsorbents for use in systems requiring a single magnetic separation, e.g., certain radioimmunoassays. Ferromagnetism limits particle usefulness in those applications requiring redispersion or reuse.
  • the magnetic core particles can be made by precipitating metal powders and reducing the particle size by milling the resulting precipitate, for example, in a ball mill.
  • the metal powder is precipitated from an aqueous solution of, for example, Fe +2 or Fe +3 salt with sodium borohydride.
  • an aqueous solution of ferrous chloride (FeCl 2 ) is mixed with sodium borohydride
  • the magnetic metal powder is then collected by filtration and washed with about five equal volumes of water to remove all soluble salts, then washed with five equal volumes of dry in acetone to remove all residual water.
  • the particle is added as an aqueous slurry in a concentration of about 1-25% to a c omme rc i al ball mill filled half way with 1/4" stainless steel balls and milled for 3-30 days.
  • a superparamagnetic metal slurry is formed and coated and functionalized as the superparamagnetic particles described in the previous section.
  • the magnetic core particles are made by reacting a metallocene, e.g., particulate ferrocene (dicyclopentadenyliron, C 10 H 10 Fe) with iron (II) hydroxide.
  • a metallocene e.g., particulate ferrocene (dicyclopentadenyliron, C 10 H 10 Fe)
  • iron (II) hydroxide iron (II) hydroxide.
  • an aqueous ferrocene (or other metallocene) slurry is prepared, and an aqueous slurry of Iron (II) hydroxide is prepared separately.
  • the ferrocene slurry is prepared, for example, by milling a mixture of ferrocene and water in a ball mill.
  • the iron (II) hydroxide slurry can be prepared, for example, by precipitating an aqueous solution of ferrous sulfate with ammonium hydroxide to form ferrous hydroxide. The two slurries are then combined and milled, for example, forming fine magnetite particles. Other metallocene compounds (e.g., nickelocene, cobaltocene) can be mixed with the ferrocene to produce various magnetic ferrite particles. This process is described in detail in co-pending U.S. patent application Serial No. _________,
  • the coating around the magnetic core particle is amino-pro- pyl- titanium- tri- isopropoxide.
  • the polymerization is performed by redispersing the magnetic particle in an acetone solution, adding the organo-titanium monomer, then crosslinking with heat.
  • Coupled magnetically responsive particle or “coupled magnetic particle” refer to any magnetic particle to which one or more types of bioaffinity adsorbents are coupled by covalent bonds, which covalent bonds may be amide, ester, ether sulfonamide, disulfide, azo or other suitable organic linkages depending on the functionalities available for bonding on both the coating of the magnetic particle and the bioaffinity adsorbents.
  • Preferred magnetically responsive particles of the present Invention have metal oxide cores composed of clusters of superparamagnetic crystals affording efficient separation of the particles in low magnetic fields (100-1000 Oersteads) while maintaining super- paramagnetic properties.
  • Aggregation of particles is controlled during particle synthesis to produce particles which are preferably small enough to avoid substantial gravitational settling over times sufficient to permit dispersions of the particles to be used in an intended biological assay or other application.
  • the advantage of having superparamagnetic cores in magnetically responsive particles is that such particles can be repeatedly exposed to magnetic fields. Superparamagnetic particles do not exhibit reminent magnetization and have no coercive strength, and, therefore, do not ma gne ti c al ly aggregate, thus, the particles can be redispersed and reused.
  • preferred particles of the invention having cores made up of clusters of crystals exhib it a remarkably high surface area per unit weight and a generally corresponding high coupling capacity, which indicates that such particles have an open or porous structure.
  • bioaffinity adsorbents can be covalently bonded to the organo-metallic coated magnetic particles of this invention by conventional coupling chemistries. Several coupling reactions can be performed. For example:
  • the ligand to be coupled contains an amino group, it can be coupled directly to the activated organo-metallic polymer. If a different functionality is desired, it can be introduced, for example, by adding a spacer arm containing the functionality by sequential reaction of the organo-metallic polymer (e.g., titanium- tetra-isopropoxide) with any omega-functional higher molecular weight alcohol. The amino group on the ligand can then be coupled to the free functional group on the spacer arm; or
  • the ligand contains an aldehyde group instead of an amino group, it can be coupled directly to the free amino group of an amino alkane (that is, an alkane spacer arm having an amino functionality) on the coated magnetic particle.
  • bioaffinity adsorbent is defined as any biological or other organic molecule capable of specific or nonspecific binding or interaction with another biological molecule, which binding or interaction may be referred to as "ligand/ligate " binding or interaction and is exemplified by, but not limited to, antibody/antigen, antibody/hapten, enzyme/substrate, carrier protein/substrate, lectin/carbohydrate, receptor/hormone, receptor/ effector or repressor/inducer bindings or interactions.
  • the coupled organo-metallic coated magnetic particles of the present invention can be used in immunoassays or other binding assays for the measurement of analytes in solution.
  • immunoassay is defined as any method for measuring the concentration or amount of an analyte in a solution based on the immunological binding or interaction of a polyclonal or monoclonal antibody and an antigen, which method (a) requires a separation of bound from unbound analyte; (b) employs a radioisotopic, fluorometrlc, enzymatic, chemiluminescent or other label as the means for measuring the bound and/or unbound analyte; and (c) may be described as
  • Immunoassays are exemplified by, but are not limited to, radioimmunoassays (RIA), immunoradiometric assays (IRMA), fluoroimmunoassays (FIA), enzyme immunoassays (EIA), and sandwich method Immunoassays.
  • the analyte or the bioaffinity adsorbent can include, for example, antibodies, antigens, haptens, enzymes, apoenzymes, enzymatic substrates, enzymatic inhibitors, cofactors, nucleic acids, binding proteins, carrier proteins, compounds bound by binding proteins, compounds bound by carrier proteins, lectins, monosaccharides, polysaccharides, hormones, receptors, repressors and inducers.
  • Such assays are preferably carried out by mixing a sample containing an unknown concentration of analyte with a known amount of labeled analyte in the presence of magnetic particles coupled to a bioaffinity adsorbent capable of binding to, or interacting with, both unlabeled and labeled analyte, allowing the binding or interaction to occur, magnetically separating the particles, measuring the amount of label associated with the magnetic particles and comparing the amount of label to a standard curve to determine the concentration of analyte in the sample.
  • binding assay or “non-immune assay” is defined as any method for measuring the concentration or amount of an analyte in solution based on the specific or nonspecific binding or interaction, other than antibody/ antigen binding or interaction, or a bioaffinity adsorbent and another biological or organic molecule, which method (a) requires a separation of bound from unbound analyte; (b) employs a radioisotopic, fluorometric, enzymatic, chemiluminescent or other label as as the means for measuring the bound and/or unbound analyte; and (c) may be described as “competitive” if the amount of bound measurable label is generally inversely proportional to the amount of analyte originally in solution or “non-competitive” if the amount of bound measurable label is generally originally in solution.
  • the magnetic organo -metallic - coated particles of this invention are useful in immobilized enzyme systems, particularly where enzyme recycling is desired.
  • immobilized enzyme system is defined as any enzymatically catalyzed biochemical conversion or synthesis or degradation wherein the enzyme molecule or active site thereof is not freely soluble but is adsorptively or covalently bound to a solid phase support, which support is suspended in or contacted with the surrounding medium and which may be reclaimed or separated from said method.
  • enzymatic reactions are carried out by dispersing enzyme- coupled magnetic particles in a reaction mixture containing one or more substrates, under conditions sufficient for the reaction between the enzyme and substrate to occur, magnetically separating the enzyme-magnetic particle from the reaction mixture containing products and unreacted substrates and, if desired, redispersing the particles in fresh substrates thereby reusing the enzyme.
  • affinity chromatography is defined as a method for separating, isolating, and/or purifying a selected molecule from its surrounding medium on the basis of its binding or interaction with a bioaffinity adsorbent adsorptively or covalently bound to a solid phase support, which support is suspended in or contacted with the surrounding medium and which may be reclaimed or separated from said medium by dispersing bioaffinity adsorbent coupled magnetic particles in solutions or suspensions containing molecules or cells to be isolated and/or purified, allowing the bioaffinity adsorbent and the desired molecules or cells to interact, magnetically separating the particles from the solutions or suspension and recovering the isolated molecules or cells from the magnetic particles.
  • the organo-metallic coated magnetic particles of this invention can be used in in vivo systems for the diagnostic localization of cells or tissues recognized by the particular bioaffinity adsorbent coupled to the particle and also for magnetically directed delivery of therapeutic agents coupled to the particles to pathological sites.
  • Magnetic separation times of less than about ten minutes can be achieved with magnetic particles of the invention by contacting a vessel containing a dispersion of the particles with a pole face of a permanent magnet no larger in volume than the volume of the vessel.
  • Magnetic separation time is defined to be the time for the turbidity of the dispersion to fall by 95 percent.
  • Example 1 Preparation of Superparamagnetic Magnetite Particles 200 grams (1.58 moles) of ferrous chloride (VWR Scientific) and 325 grams (2.0 moles) of ferric chloride were dissolved in 3 liters of water. 2000 grams of ammonium hydroxide (VWR Scientific) concentrate were added at a rate of 50 ml/minute under constant agitation, during which time the temperature of the solution was kept between 25 and 40°C. After the addition of the ammonium hydroxide was complete, the magnetic particle (Fe 3 O 4 ) aqueous slurry was allowed to cool to room temperature.
  • VWR Scientific ferrous chloride
  • VWR Scientific ammonium hydroxide
  • Example #3 The material was cooled to room temperature and used as a monomer in making the tetravalent titanium organo- metallic coating in Example #3.
  • Example 2 Preparation of Amine Functional Organo- titanate Coated Magnetic Particle According to the procedure s et out in Exampl e 1 , 4 moles FeCl 3 and 2 moles of FeCl 2 were dissolved in 4 L of distilled water and precipitated with 16 moles of ammonium hydroxide. The precipitate was washed 5 time s with water and 3 times with acetone. N,N-dimethyl formamlde (DMF) was added to the precipitate in the following ratio: 10 ml of DMF per gram of Fe 3 O 4 . The mixture was loaded into a Eiger Mill and mill e continuously for 10 minutes. The mixture was then transferred to a beaker and heated with stirring for 30 minutes at 100°C. The amine functional organo-titanate prepared in Example 2 was immediately added after preparation with constant stirring to the mixture in a ratio of 1 g dry Fe 3 O 4 per 3 g of amine functional organo-titanate.
  • DMF N,N-dimethyl formaml
  • Example 2 The procedure described in Example 2 was fo l lowe d except that the organo-titanate was reacted with a comixture of amino-functional hexanol and hexanol to produce a monomer having reduced amine functionality.
  • Hexanol and 6 -amino- 1 -hexanol in a molar ratio of 6:1 were mixed in a 50 ml beaker for one minute.
  • Tyzor TPT was added to the alcohol mixture in the ratio of 1 mole of alcohol per mole of Tyz or TPT.
  • the reaction mixture was s tirred for one minute , he ated to 70 ° C fo r 10 minutes to evaporate the isopropyl alcohol produced by the reaction and cooled to room temperature.
  • the resulting compound was an organotitanate, 6 - amino -hexyl-titanium- tri-isopropoxide having alternating non- functional hexyl groups, that is, hexyl chains lacking the amino group.
  • the weight ratios of 6 - amino - 1 -hexanol: Tyzor TPT:hexanol were 1:26:9.6.
  • This compound was used as a monomer to make an organo - titanium coating as described in Example 5.
  • Example 3 The procedure described in Example 3 was followed except that the amine-functional organo-titanate was the material prepared in Example 5.
  • the mixture of magnetic particles and organo-titanate monomer was heated to 95°C for one hour with constant stirring and milled in an
  • Eiger Mill for 4 minutes. The mixture was washed nine times with water. Adipic acid was added in the ratio of 0.5 moles of adipic acid per mole of total particles. One mole of carbodiimide (CDI) was added, and the mixture was mixed for 30 minutes on a ball mill. 1,6 hexane- diamine was added in the ratio of 0.5 moles of 1,6 hexane-diamine per mole of total particles. One mole of CDI was added and the mixture was mixed for 30 minutes. The resulting material was washed five times with water, the particles were collected using an external magnetic field of 2000 gauss and the aqueous waste was decanted.
  • CDI carbodiimide
  • Example 6 Preparation of Subdomain Magnetite Particles by Reaction of Particulate Ferrocene and Iron (II) Hydroxide
  • a slurry containing 20% ferrocene (by weight) (dicyclopentadenyliron; Strem Chemical Co., Newburyport, MA) in water was prepared by mixing the ferrocene with the water.
  • the slurry was added to a commercial ball mill. The mill was filled halfway with 1 ⁇ 4 " stainless steel balls and the slurry was milled for a period of 2 hours.
  • a second ferrous hydroxide slurry (iron (II) hydroxide) was made according to the following procedure.
  • An aqueous solution containing 20g of ferrous sulfate (VWR Scientific) was precipitated using 50g of ammonium hydroxide concentrate to form gelatinous ferrous hydroxide.
  • the gel was filtered and the filtrate washed with 5 to 100g volumes of water. The washed gel was then made into a 10% aqueous slurry and milled as previously described for 5 hours.
  • the ferrocene and hydroxide slurries were mixed, and the mixture was milled for one day to form fine Fe 3 O 4 particles.
  • the particles were about 100 A in diameter and were responsive to a magnetic field. These particles can be coated as described in Examples 2-5 above.
  • Example 7 Preparation of Subdomain Nickel-Ferrite Particles
  • Subdomain nickel-ferrite particles were prepared according to the procedure set out in Example 6, except that a mixture of 50g a 20% nickelocene slurry (dicyclopentadenylnickel; Strem Chemical Co., Newburyport, MA) and 50g of a 20% ferrocene slurry were used in lieu of the 100g of the ferrocene slurry in Example 6.
  • Magnetically responsive nickel-ferrite particles having a particle size of about 100 A were produced by this method.
  • Example 8 Preparation Subdomain Cobat- Ferrite Particles
  • Subdomain cobalt - ferrite particles were prepared according to the procedure set out in Example 6, except that a mixture of 50g of a 20% (by wt.) cobaltocene slurry (dicyclopentadenylcobalt; Strem Chemical Co., Newburyport, MA ) and 50g of the ferrocene slurry were used in lieu of 100g of the ferrocene slurry in Example 6.
  • Magnetically responsive cobalt-ferrite particles having a particle size of about 100 A were produced by this method.
  • Example 9 Preparation of Subdomain Metal Particles by Sodium Borohydride Reduction and Size Reduction by Milling 200 gm (1.58 moles) of ferrous chloride was dissolved in 1 liter of water. 500 gm of dry sodium borohydride were added to the solution to form a fine iron powder precipitate. The precipitate was washed with water and collected by filtration. The filtered powder was resuspended in water and re-filtered. The washing procedure was done 4 additional times. On the final suspension, the slurry was adjusted to a concentrate of 20% and milled as described in Example 6 for a period of 75 days to produce particles with a mean diameter of less than 50 A.

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Abstract

L'invention concerne des particules à sensibilité magnétique ainsi que leur utilisation dans des systèmes où la séparation de certaines molécules du milieu environnant est nécessaire ou désirable. Les particules magnétiques consistent en un noyau métallique, d'oxyde métallique ou d'alliage métallique revêtues d'un polymère organo-métallique auquel est rattachée une fonctionnalité organique à laquelle une variété de molécules organiques et/ou biologiques peuvent être couplées. Les particules peuvent être dispersées dans des milieux aqueux sans décantation rapide par gravité et récupérées des milieux aqueux en utilisant un champ magnétique. Les particules à sensibilité magnétique de l'invention peuvent être couplées à des molécules biologiques ou organiques avec une affinité pour certaines autres molécules biologiques ou organiques, ou avoir la capacité d'en adsorber ou pouvant avoir une interaction avec certaines autres molécules biologiques ou organiques. Des particules ainsi couplées peuvent être utilisées dans une variété de systèmes in vitro ou in vivo impliquant des étapes de séparation ou le mouvement dirigé des molécules couplées vers des sites particuliers, y compris des analyses immunologiques, d'autres analyses biologiques, des réactions biochimiques ou enzymatiques, des purifications chromatographiques par affinité, le triage de cellules et des utilisations diagnostiques et thérapeutiques.
PCT/US1990/007492 1989-12-22 1990-12-18 Particules revetues d'un polymere organo-metallique utilisees dans des separations WO1991009678A1 (fr)

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Cited By (18)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0546939A1 (fr) * 1991-12-13 1993-06-16 Molecular Bioquest, Inc. Micrograppes magnétiques et méthodes de préparations
WO1993026019A1 (fr) * 1992-06-08 1993-12-23 Molecular Bioquest, Inc. Preparation de particules inorganiques a dimensions reglables utiles dans des separations, comme commutateurs moleculaires magnetiques et comme liposomes inorganiques dans des applications medicales
US5389377A (en) * 1989-12-22 1995-02-14 Molecular Bioquest, Inc. Solid care therapeutic compositions and methods for making same
US5441746A (en) * 1989-12-22 1995-08-15 Molecular Bioquest, Inc. Electromagnetic wave absorbing, surface modified magnetic particles for use in medical applications, and their method of production
US5571531A (en) * 1994-05-18 1996-11-05 Mcmaster University Microparticle delivery system with a functionalized silicone bonded to the matrix
WO1999021934A1 (fr) * 1997-10-28 1999-05-06 The University Of Melbourne Particules stabilisees et procedes de et d'utilisation desdites particules
EP0928611A1 (fr) * 1996-06-10 1999-07-14 Nittetsu Mining Co., Ltd. Poudre medicale
US5935866A (en) * 1989-12-22 1999-08-10 Binax Nh, Inc. Preparation of sub 100 A magnetic particles and magnetic molecular switches
WO1999059713A1 (fr) * 1998-05-18 1999-11-25 Studiengesellschaft Kohle Mbh Procede pour modifier les proprietes de dispersion de colloides de nanoparticules metalliques prestabilises ou pretraites par voie organo-metallique
WO2000061191A2 (fr) * 1999-04-09 2000-10-19 Advanced Magnetics, Inc. Oxydes de fer colloidaux thermostables recouverts de carbohydrates reduits et des derives de carbohydrates
US6468808B1 (en) 1998-09-24 2002-10-22 Advanced Research And Technology Institute, Inc. Water-soluble luminescent quantum dots and biomolecular conjugates thereof and related compositions and method of use
EP1522318A3 (fr) * 1999-04-09 2005-07-06 Advanced Magnetics Incorporated Oxydes de fer colloidaux thermostables enrobés
US7592182B2 (en) 2004-08-03 2009-09-22 Axis-Shield Diagnostics Limited Assay
US7871597B2 (en) 1999-04-09 2011-01-18 Amag Pharmaceuticals, Inc. Polyol and polyether iron oxide complexes as pharmacological and/or MRI contrast agents
JP2013526701A (ja) * 2010-05-10 2013-06-24 バイオ−レイヤー プロプライエタリー リミテッド 結合系
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CN103217529A (zh) * 2013-03-17 2013-07-24 南昌大学 一种基于γ-Fe2O3纳米粒子间接富集免疫磁分离的食源性致病菌快速检测方法
CN111841881A (zh) * 2020-07-29 2020-10-30 攀钢集团矿业有限公司 选矿过程中回收钛磁铁矿的方法

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US8119162B2 (en) 2005-11-10 2012-02-21 Colgate-Palmolive Company Particles that disrupt or impede bacterial adhesion, related compositions and methods
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JP6479799B2 (ja) * 2013-08-13 2019-03-06 アンテオ テクノロジーズ プロプライエタリー リミテッドＡｎｔｅｏ Ｔｅｃｈｎｏｌｏｇｉｅｓ Ｐｔｙ Ｌｔｄ 粒子への分子の接合

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0075346A1 (fr) * 1981-09-18 1983-03-30 Océ-Nederland B.V. Poudre développatrice colorée, procédé pour sa préparation, et procédé de développement
EP0125995A2 (fr) * 1983-05-12 1984-11-21 Advanced Magnetics, Inc. Particules magnétiques pour l'utilisation dans des séparations
EP0180384A2 (fr) * 1984-11-01 1986-05-07 TECHNICON INSTRUMENTS CORPORATION(a Delaware corporation) Porteur-réactif sensible au magnétisme et procédé de préparation
EP0260098A2 (fr) * 1986-09-06 1988-03-16 Bellex Corporation Conjugat de substance physiologiquement active et de matériau magnétique
EP0321322A1 (fr) * 1987-12-18 1989-06-21 Rhone-Poulenc Chimie Particules composites magnétisables à base d'organopolysiloxane réticulé, leur procédé de préparation et leur application en biologie

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0075346A1 (fr) * 1981-09-18 1983-03-30 Océ-Nederland B.V. Poudre développatrice colorée, procédé pour sa préparation, et procédé de développement
EP0125995A2 (fr) * 1983-05-12 1984-11-21 Advanced Magnetics, Inc. Particules magnétiques pour l'utilisation dans des séparations
EP0180384A2 (fr) * 1984-11-01 1986-05-07 TECHNICON INSTRUMENTS CORPORATION(a Delaware corporation) Porteur-réactif sensible au magnétisme et procédé de préparation
EP0260098A2 (fr) * 1986-09-06 1988-03-16 Bellex Corporation Conjugat de substance physiologiquement active et de matériau magnétique
EP0321322A1 (fr) * 1987-12-18 1989-06-21 Rhone-Poulenc Chimie Particules composites magnétisables à base d'organopolysiloxane réticulé, leur procédé de préparation et leur application en biologie

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Chemical Abstacts, vol. 108, no. 10, 7 March 1988, (Columbus, Ohio, US), see page 318 *
Chemical Abstracts, vol. 89, no. 26, 25 December 1978, (Columbus, Ohio, US), see page 656 *

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Publication number Priority date Publication date Assignee Title
US5389377A (en) * 1989-12-22 1995-02-14 Molecular Bioquest, Inc. Solid care therapeutic compositions and methods for making same
US5441746A (en) * 1989-12-22 1995-08-15 Molecular Bioquest, Inc. Electromagnetic wave absorbing, surface modified magnetic particles for use in medical applications, and their method of production
US5935866A (en) * 1989-12-22 1999-08-10 Binax Nh, Inc. Preparation of sub 100 A magnetic particles and magnetic molecular switches
US5382468A (en) * 1991-12-13 1995-01-17 Molecular Bioquest, Inc. Biodegradable magnetic microclusters and methods for making them
EP0546939A1 (fr) * 1991-12-13 1993-06-16 Molecular Bioquest, Inc. Micrograppes magnétiques et méthodes de préparations
WO1993026019A1 (fr) * 1992-06-08 1993-12-23 Molecular Bioquest, Inc. Preparation de particules inorganiques a dimensions reglables utiles dans des separations, comme commutateurs moleculaires magnetiques et comme liposomes inorganiques dans des applications medicales
US5571531A (en) * 1994-05-18 1996-11-05 Mcmaster University Microparticle delivery system with a functionalized silicone bonded to the matrix
EP0928611A4 (fr) * 1996-06-10 1999-12-15 Nittetsu Mining Co Ltd Poudre medicale
EP0928611A1 (fr) * 1996-06-10 1999-07-14 Nittetsu Mining Co., Ltd. Poudre medicale
US6162469A (en) * 1996-06-10 2000-12-19 Nittetsu Mining Co., Ltd. Medical powder
US6548168B1 (en) 1997-10-28 2003-04-15 The University Of Melbourne Stabilized particles and methods of preparation and use thereof
WO1999021934A1 (fr) * 1997-10-28 1999-05-06 The University Of Melbourne Particules stabilisees et procedes de et d'utilisation desdites particules
US6531304B1 (en) * 1998-05-18 2003-03-11 Studiengesellschaft Kohle Mbh Method for modifying the dispersion characteristics of metal organic-prestabilized or pre-treated nanometal colloids
WO1999059713A1 (fr) * 1998-05-18 1999-11-25 Studiengesellschaft Kohle Mbh Procede pour modifier les proprietes de dispersion de colloides de nanoparticules metalliques prestabilises ou pretraites par voie organo-metallique
US7192785B2 (en) 1998-09-24 2007-03-20 Indiana University Research And Technology Corporation Water-soluble luminescent quantum dots and biomolecular conjugates thereof and related compositions and methods of use
US6468808B1 (en) 1998-09-24 2002-10-22 Advanced Research And Technology Institute, Inc. Water-soluble luminescent quantum dots and biomolecular conjugates thereof and related compositions and method of use
WO2000061191A3 (fr) * 1999-04-09 2001-05-25 Advanced Magnetics Inc Oxydes de fer colloidaux thermostables recouverts de carbohydrates reduits et des derives de carbohydrates
US6599498B1 (en) 1999-04-09 2003-07-29 Advanced Magnetics, Inc. Heat stable colloidal iron oxides coated with reduced carbohydrates and carbohdrate derivatives
EP1522318A3 (fr) * 1999-04-09 2005-07-06 Advanced Magnetics Incorporated Oxydes de fer colloidaux thermostables enrobés
US8501158B2 (en) 1999-04-09 2013-08-06 Amag Pharmaceuticals, Inc. Polyol and polyether iron oxide complexes as pharmacological and/or MRI contrast agents
US7553479B2 (en) 1999-04-09 2009-06-30 Amag Pharmaceuticals, Inc. Heat stable colloidal iron oxides coated with reduced carbohydrates and uses thereof
WO2000061191A2 (fr) * 1999-04-09 2000-10-19 Advanced Magnetics, Inc. Oxydes de fer colloidaux thermostables recouverts de carbohydrates reduits et des derives de carbohydrates
EP1169062B1 (fr) 1999-04-09 2009-10-21 AMAG Pharmaceuticals, Inc. Oxydes de fer colloidaux thermostables recouverts de carbohydrates reduits et des derives de carbohydrates
US7871597B2 (en) 1999-04-09 2011-01-18 Amag Pharmaceuticals, Inc. Polyol and polyether iron oxide complexes as pharmacological and/or MRI contrast agents
US9555133B2 (en) 1999-04-09 2017-01-31 Amag Pharmaceuticals, Inc. Polyol and polyether iron oxide complexes as pharmacological and/or MRI contrast agents
US8926947B2 (en) 1999-04-09 2015-01-06 Amag Pharmaceuticals, Inc. Polyol and polyether iron oxide complexes as pharmacological and/or MRI contrast agents
US8591864B2 (en) 1999-04-09 2013-11-26 Amag Pharmaceuticals, Inc. Polyol and polyether iron oxide complexes as pharmacological and/or MRI contrast agents
US7592182B2 (en) 2004-08-03 2009-09-22 Axis-Shield Diagnostics Limited Assay
JP2013526701A (ja) * 2010-05-10 2013-06-24 バイオ−レイヤー プロプライエタリー リミテッド 結合系
CN103217529A (zh) * 2013-03-17 2013-07-24 南昌大学 一种基于γ-Fe2O3纳米粒子间接富集免疫磁分离的食源性致病菌快速检测方法
CN103185796A (zh) * 2013-03-17 2013-07-03 南昌大学 一种基于γ-Fe2O3@Au纳米粒子间接富集免疫磁分离的食源性致病菌快速检测方法
CN111841881A (zh) * 2020-07-29 2020-10-30 攀钢集团矿业有限公司 选矿过程中回收钛磁铁矿的方法

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