EP1934609A4 - Nanoparticules magnétiques présentant une fluorescence, leur procédé de préparation et leur utilisation - Google Patents

Nanoparticules magnétiques présentant une fluorescence, leur procédé de préparation et leur utilisation

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Publication number
EP1934609A4
EP1934609A4 EP06798700A EP06798700A EP1934609A4 EP 1934609 A4 EP1934609 A4 EP 1934609A4 EP 06798700 A EP06798700 A EP 06798700A EP 06798700 A EP06798700 A EP 06798700A EP 1934609 A4 EP1934609 A4 EP 1934609A4
Authority
EP
European Patent Office
Prior art keywords
magnetic nanoparticles
nanoparticles
magnetic
solution
bound
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP06798700A
Other languages
German (de)
English (en)
Other versions
EP1934609A1 (fr
Inventor
Jin-Kyu Lee
Myung-Haing Cho
Seung-Bum Park
Tae-Jong Yoon
Jun-Sung Kim
Byung-Geol Kim
Kyeong-Nam Yu
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BITERIALS Co Ltd
Original Assignee
BITERIALS CO Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from KR1020050112245A external-priority patent/KR100821192B1/ko
Application filed by BITERIALS CO Ltd filed Critical BITERIALS CO Ltd
Publication of EP1934609A1 publication Critical patent/EP1934609A1/fr
Publication of EP1934609A4 publication Critical patent/EP1934609A4/fr
Withdrawn legal-status Critical Current

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Classifications

    • 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
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/588—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with semiconductor nanocrystal label, e.g. quantum dots
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y15/00—Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
    • 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

Definitions

  • the present invention relates to magnetic nanoparticles (MNPs) having fluorescence, and preparation and use thereof.
  • MNPs magnetic nanoparticles
  • nanoparticles including quantum dots are composed of heavy metals such as cadmium (Cd), zinc (Zn), cobalt (Co) and the like, surfaces of the synthesized nanoparticles should be made biocompatible, in order to enhance the applicability thereof to bio-fields.
  • the present invention has been made in view of the above problems, and it is an object of the present invention to provide magnetic nanoparticles having fluorescence.
  • the inventors of the present invention have investigated a method of modifying a surface of magnetic nanoparticles, which contain organic fluorescent materials and are coated with silica shells, with an electrically charged material.
  • a method of modifying a surface of magnetic nanoparticles which contain organic fluorescent materials and are coated with silica shells, with an electrically charged material.
  • we have synthesized magnetic nanoparticles coated with silica containing the organic fluorescent materials and having a surface modified with the electrically charged material and have confirmed that upon introduction of such magnetic nanoparticles into cells, it is possible to locate and control the introduced nanoparticles by application of an external magnetic field, and simultaneously it is also possible to efficiently apply such particles to both in vivo and in vitro studies via easy and convenient detection of the fluorescence.
  • the present invention has been completed based on these findings.
  • the above and other objects can be accomplished by the provision of magnetic nanoparticles having a core containing a magnetic material and a surface-modified silica shell containing an organic fluorescent material and coated on the core, wherein the nanoparticles have a size of less than 100 nm and are water-soluble.
  • magnetic nanoparticles wherein the magnetic nanoparticles are bound to negatively charged genes or nucleic acid molecules, and a gene delivery system comprising the same.
  • magnetic nanoparticles wherein magnetic nanoparticles having fluorescence are bound to negatively charged nucleic acid molecules, and a gene delivery system comprising the same.
  • magnetic nanoparticles wherein magnetic nanoparticles having fluorescence are bound to antibodies, and a cell staining agent comprising the same.
  • Magnetic nanoparticles according to the present invention have both optical and magnetic properties and are applicable to bio-fields.
  • chemical functional groups can be introduced into nano-scale materials, using a variety of compounds.
  • Use of the thus-chemically modified nano-scale materials can lead to an increased or decreased penetrability of magnetic nanoparticles into cells.
  • the magnetic nanoparticles can be usefully used as a gene delivery system by transfer of a desired plasmid DNA into a target cell using nano-scale materials having a positive charge, and can also be usefully used in cell staining, based on a technique which is capable of performing selective binding of nanoparticles to certain cells and recognition of the nanoparticle-bound cells, using an appropriate surface treatment technique.
  • the selectively recognized cells can be separated and purified by application of a strong external magnetic field.
  • FIG. 1 is a view showing a process for preparing magnetic nanoparticles
  • FIGS. 2A to 2C are transmission electron micrographs (TEMs) of magnetic nanoparticles (MNP @ SiO (RITC or FITC)) containing organic fluorescent materials and coated with silica shells;
  • FIG. 3 is a view showing a chemical treatment process of a surface of magnetic nanoparticles (MNP @ SiO (RITC)) according to the present invention with various silicon compounds;
  • FIG. 4 is a graph showing zeta-potential for the measurement of changes in a surface charge of all the magnetic nanoparticles, due to various surface treatments of magnetic nanoparticles (MNP@SiO (RITC)) according to the present invention
  • Black Non-surface treated MNP@SiO (RITC)
  • Red (CH O) Si-PEG-surface treated MNP @ SiO (RITC)-PEG
  • Light green (CH O) Si-PMP-surface treated MNP @ SiO (RITC)-PMP
  • Blue (CH O) Si-PTMA-surface treated MNP@SiO (RITC)-PTMA
  • FIGS. 5A to 5D are confocal laser scanning micrographs showing a penetration rate of MNP @ SiO (RITC)-PEG, MNP @ SiO (RITC)-PTMA, MNP @ SiO (RITC) and MNP @ SiO (RITC)-PMP into breast cancer cells;
  • FIGS. 6 A to 6H are confocal laser scanning micrographs showing intracellular location of nanoparticles, upon injection of MNP@Si0 (RITC)-PEG and MNP@Si0 (RITC)-PMP into breast cancer cells at the same amounts under the same conditions
  • 6A to 6D Micrographs for injection of MNP@Si0 (RITC)-PEG
  • 6E to 6H Micrographs for injection of MNP @ SiO (RITC)-PMP
  • 6A and 6E Red fluorescence micrographs
  • 6B and 6F Optical micrographs
  • 6C and 6G Fluorescence micrographs confirming DAPI nuclear staining
  • 6D and 6F Overlapping micrographs of 6A to 6C and 6E to 6G, respectively);
  • FIG. 7 is a bar graph showing results of cytotoxic test (MTT assay) after treatment of MNP@SiO (RITC), MNP@SiO (RITC)-PEG, MNP@SiO (RITC)-PMP, and MNP @ SiO (RITC)-PTMA on a breast cancer cell line (MCF-7), a lung cancer cell line (A549) and a normal (non-malignant) lung epithelial cell line (NL20), respectively;
  • FIG. 8 is a view showing a process using MNP @ SiO (RITC)-PTMA as a gene delivery system, by binding of it to a plasmid DNA;
  • FIGS. 9 A to 9D are confocal laser scanning micrographs of transfected cells after gene delivery using plasmid DNA-bound MNP @ SiO (RITC)-PTMA (9A: Blue fluorescence micrograph, 9B: Optical micrograph, 9C: Red fluorescence micrograph, and 9D: Overlapping micrograph of 9 A, 9B and 9C);
  • FIG. 10 is a view showing a process of co-treating a surface of MNP@Si0 (FITC) with (CH O) Si-PEG and 3-aminopropyltriethoxysilane (APS), introducing a maleimide group into an amine group of the MNP @ SiO (FITC) surface, and introducing an antibody for recognition of a certain cell thereto;
  • FITC MNP@Si0
  • APS 3-aminopropyltriethoxysilane
  • FIGS. 1 IA to 1 ID are confocal laser scanning micrographs showing utilization of antibody-bound MNP@Si0 (FITC)-PEG/APS-MaI in cell staining (HA: Blue fluorescence micrograph, HB: Optical micrograph, HC: Red fluorescence micrograph, and 1 ID: Overlapping micrograph of 1 IA, 1 IB and HC); wherein a material penetrated into cells is MNP @ SiO (RITC) emitting red fluorescence, and a cell membrane-bound material is MNP @ SiO (FITC)-PEG/APS-MaI-Her2 emitting blue
  • MNP @ SiO (FITC)-Her2 Ab fluorescence
  • FIGS. 12A to 12F are micrographs showing a selectivity of MNP@SiO
  • FITC-CDlO having a CD-IO antibody, being capable of selectively binding to a
  • FIGS. 13A and 13B are optical micrographs showing that MNP@SiO
  • FITC-CDlO is selectively recognized by a cell wall of a leukemia cell and is then
  • FIG. 14 shows results of MRI analysis at predetermined time intervals after intraperitoneal injection of MNP @ SiO (RITC) into mice, wherein a control is a micrograph of a mouse with no injection of magnetic nanoparticles, and the remainder are micrographs taken 15 min, 30 min, 1 hour, 1 day and 3 days after synthesized magnetic nanoparticles were injected into mice. Best Mode for Carrying Out the Invention
  • Magnetic nanoparticles of the present invention contain a magnetic material inside the particle and the outside of the core thereof is coated with a non-magnetic silica shell containing an organic fluorescent material and having a surface modified with an electrically charged material. Therefore, the magnetic nanoparticles of the present invention have both optical and magnetic properties and can be applied to a variety of bio-fields.
  • the magnetic nanoparticles of the present invention can be prepared by a method comprising the steps of:
  • Step 3 with a silicon compound.
  • the water-soluble magnetic nanoparticles may be prepared according to any conventional method known in the art, such as wet, dry or vacuum method. Examples of such a method may include, but are not limited to, grinding of large size materials, precipitation from a solution, co-precipitation, microemulsification, polyol process, high-temperature degradation of organic precursors, solution techniques, aerosol/bubble methods, spray pyrolysis, plasma atomization and laser pyrolysis.
  • the water-soluble magnetic nanoparticles of the present invention may be prepared by co-precipitation.
  • the water-soluble magnetic nanoparticles are composed of cobalt (Co) and iron
  • (Fe) oxides may include an oxide of a transition metal such as manganese (Mn), zinc (Zn), nickel (Ni), copper (Cu) or the like.
  • the organic fluorescent material is preferably Rhodamine B isoth- iocyanate (RITC) or fluoresceine isothiocyanate (FITC), but is not limited thereto and may include chemical modifications of the existing organic fluorescent materials.
  • Rhodamine B isoth- iocyanate FITC
  • FITC fluoresceine isothiocyanate
  • the silicon compound used in the surface modification of the silica shell is preferably an electrically charged material, i.e. an organic silicon compound having ionic functional group(s).
  • an organic silicon compound may include specific functional compounds such as ionic compounds, water soluble compounds and drugs, to which a (CH O) Si- functional group is introduced.
  • the silicon compound that can be used in the present invention may include, but is not limited to, one compound selected from the group consisting of (CH O) Si-PEG [(CH O) SiCH CH 2 CH 20(CH 2 CH 2 O) 6-9 CH 3 ], (CH 3 O) 3 Si-PMP [(CH 3 O) 3 SiCH 2 CH 2 CH 2 PO 2 (OCH 3 )Na],
  • the magnetic nanoparticles according to the present invention exhibit no toxicity for all kinds of cells including a breast cancer cell line (MCF-7), a lung cancer cell line (A549) and a normal (non-malignant) lung epithelial cell line (NL20).
  • MCF-7 breast cancer cell line
  • A549 lung cancer cell line
  • NL20 normal (non-malignant) lung epithelial cell line
  • the number of the magnetic nanoparticles is not sufficient to cause cytotoxicity, but is enough to provide cells exhibiting magnetically induced movement.
  • the above-mentioned cell may include eukaryotic cells, human cells, animal cells and plant cells.
  • the magnetic particles may have an average particle size of less than about 100 nm, particularly about 30 to 80 nm.
  • the cells to which the magnetic nanoparticles were penetrated are moved at a rate of 0.5 to 1 mm/sec by application of the external magnetic field (ca. 0.3 tesla, T).
  • the external magnetic field strength and the moving rate are not limited to the above- specified range.
  • the magnetic nanoparticles according to the present invention coated with silica shells containing an organic fluorescent material and having a surface modified with the electrically charged material, can be used for various applications, by binding the surface of the surface-modified silica shell to various materials such as negatively charged genes or nucleic acid molecules and antibodies.
  • the magnetic nanoparticles bound to the negatively charged genes can be prepared by a method comprising the steps of:
  • Step 3 3) adding Dulbecco's Modified Eagle Medium (DMEM) to the incubated solution of Step 2, adjusting a Ca + ion concentration of the solution to 4.5 mM, further incubating the solution for 4 hours and washing the solution with a phosphate-buffered saline (PBS) solution.
  • DMEM Dulbecco's Modified Eagle Medium
  • DNA-[MNP@SiO (RITC)-PTMA] ⁇ enter the target cells, they pass through the cell membrane and deliver the negatively charged genes into the cells and are then separated from the genes and remain as the magnetic nanoparticles in the cytoplasm (red fluorescence). In addition, it can be confirmed that a blue protein was synthesized in the cytoplasm by the delivered DNA (see FIG. 8).
  • the negatively charged gene includes, but is not limited to, a plasmid
  • the magnetic nanoparticles according to the present invention may be bound to various genes.
  • the magnetic nanoparticles according to the present invention may also be bound to negatively charged nucleic acid molecules, in addition to negatively charged genes.
  • the magnetic nanoparticles according to the present invention may be usefully used as a gene delivery system, by attachment of the particles to the negatively charged genes or nucleic acid molecules.
  • the magnetic nanoparticles may be selectively bound to certain cells.
  • the magnetic nanoparticles-introduced cells may be separated via the induction of a movement thereof by application of an external magnetic field.
  • the antibody-bound magnetic nanoparticles can be prepared by a method comprising the steps of:
  • Examples of the antibody that can be used in Step 4 may include CD-10 antibody against leukemia cells and Her2 antibody against breast cancer cells.
  • the antibodies that can be used in the present invention are not limited thereto, and therefore may include antibodies of various cells including stem cells.
  • the magnetic nanoparticles according to the present invention may be usefully used as a cell staining agent, by binding of the nanoparticles to the antibodies of interest.
  • the magnetic nanoparticles according to the present invention are observed as a black magnetic signal in the liver of mice, following intraperitoneal administration of the nanoparticles into the animals.
  • the magnetic nanoparticles according to the present invention may be usefully used in cell staining (bio-imaging), cell separation, in vivo drug delivery and in vivo gene transfer.
  • the magnetic nanoparticles according to the present invention may be used as an assay reagent which is capable of simultaneously performing fluorescence analysis and MRI analysis.
  • an assay reagent which is capable of simultaneously performing fluorescence analysis and MRI analysis.
  • a solution of an organic fluorescent material e.g. RITC (Rhodamine B isoth- iocyanate) or FITC(fluoresceine isothiocyanate) treated with 3-aminopropyltriethoxysilane (APS), and a solution of tetraethoxysilane (TEOS) in ethanol (mole ratio of 0.04:0.3).
  • RITC Rhodamine B isoth- iocyanate
  • FITC(fluoresceine isothiocyanate) treated with 3-aminopropyltriethoxysilane (APS)
  • TEOS tetraethoxysilane
  • 0.86 mL of NH OH containing 30% by weight of NH was added to the mixed solution, thereby inducing the formation of silica on the surface of the magnetic nanoparticles.
  • the magnetic nanoparticles coated with the organic fluorescent material-containing silica shells were centrifuged at 18,000 rpm for 30 min, using a high-speed centrifuge, and the precipitates were purified by water and ethanol washing. The resulting material was readily dispersible in water or an alcohol.
  • FIG. 1 shows a process for preparing the magnetic nanoparticles coated with the organic fluorescent material-containing silica shells (MNP @ SiO (RITC or FITC)), and FIG. 2 shows TEMs of the magnetic nanoparticles coated with the organic fluorescent material-containing silica shells (MNP @ SiO (RITC or FITC)).
  • TEOS tetraethoxysilane
  • FIG. 3 shows a chemical treatment process of a surface of magnetic nanoparticles according to the present invention with various silicon compounds
  • FIG. 4 shows a graph of zeta-potential for the measurement of changes in a surface charge of all the magnetic nanoparticles, due to various surface treatments of magnetic nanoparticles according to the present invention.
  • MNP@SiO (RITC) Black line
  • (CH O) Si-PEG-surface treated MNP @ SiO (RITC)-PEG Red line
  • (CH O) Si-PMP-surface treated MNP@SiO (RITC)-PMP Light green
  • (CH O) Si-PTMA-surface treated MNP @ SiO (RITC)-PTMA (Blue line) exhibited a charge value of +35.7 mV.
  • MCF-7 breast cancer cell line
  • the breast cancer cell line was cultured in DMEM (Dulbecco's Modified Eagle's Medium) containing 40 D of 10% fetal bovine serum (FBS), and 2 mg/mL of non-surface treated magnetic nanoparticles [MNP@SiO (RITC)] and 2 mg/ mL of silicon-surface treated magnetic nanoparticles [MNP @ SiO (RITC)-PEG, MNP @ SiO (RITC)-PMP, or MNP @ SiO (RITC)-PTMA], prepared in Example 1.
  • DMEM Dynabecco's Modified Eagle's Medium
  • FBS fetal bovine serum
  • MNP@SiO (RITC) non-surface treated magnetic nanoparticles
  • silicon-surface treated magnetic nanoparticles MNP @ SiO (RITC)-PEG, MNP @ SiO (RITC)-PMP, or MNP @ SiO (RITC)-PTMA]
  • FIG. 6A to 6D are micrographs for injection of
  • MNP @ SiO (RITC)-PEG, and 6E to 6H are micrographs for injection of MNP @ SiO (RITC)-PMP.
  • 6A and 6E are red fluorescence micrographs
  • 6B and 6F are optical micrographs
  • 6C and 6G are fluorescence micrographs confirming DAPI nuclear staining
  • 6D and 6F are overlapping micrographs of 6A to 6C and 6E to 6G, respectively.
  • the (CH O) Si-PEG-surface treated magnetic nanoparticles have neutral electrical properties and are therefore irregularly distributed in the cytoplasm upon penetration thereof into cells
  • the (CH O) Si-PMP-surface treated magnetic nanoparticles have anionic properties and are therefore exist around the nuclear membrane.
  • the magnetic nanoparticles of the present invention coated with silica shells containing the organic fluorescent materials and having a surface modified with the electrically charged material have different locations in the cells, depending upon kinds of modifying components, it is possible to induce changes in the intracellular location of the nanoparticles by using the surface charge of the magnetic nanoparticles according to the present invention.
  • a breast cancer cell line (MCF-7), a lung cancer cell line (A549) and a normal
  • NL20 non-malignant lung epithelial cell line
  • DMEM Dulbecco's Modified Eagle's Medium
  • FBS fetal bovine serum
  • A549 cell line and the NL20 cell line were cultured in RPMI containing 10% FBS, 2 rnM L- glutamine, 1 mM sodium pyruvate, 1 non-essential amino acids and 5 mM 2-mercaptoethanol under the same culture conditions. All the cell lines were cultured in a Lab-Tek glass chamber slide to facilitate observation under a confocal laser scanning microscope (CLSM).
  • CLSM confocal laser scanning microscope
  • MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] was added to each well and phosphate-buffered saline (PBS, 0.2 mg/mL, pH 7.2) was then added to make a final MTT concentration of 0.4 mg/mL.
  • Cells were further incubated in a 5 % CO environment at 37 0 C for 4 hours. The culture medium was carefully removed by pipetting, and formazan crystals, formed by the action of the mitochondrial dehydrogenase which is responsible for cellular respiration of viable cells, were dissolved in 150 D of DMSO. The resulting solution was stirred for about 10 min using a stirrer, and an optical density (OD) was measured at 490 nm and 620 nm, respectively.
  • PBS phosphate-buffered saline
  • OD optical density
  • the magnetic nanoparticles according to the present invention exhibited no cytotoxicity for all kinds of cells [a breast cancer cell line (MCF-7), a lung cancer cell line (A549) and a normal (non-malignant) lung epithelial cell line (NL20)].
  • plasmid DNA gene pcDNA3.1/CT-GFP was used.
  • HEPES N-(2-hydroxyethyl)-piperazine-N'-(2-ethanesulfonic acid)] buffer solution (pH of ca. 7.4), and the resulting hybridization product was incubated at 4 0 C for 2 hours, followed by addition of 30 D of 100 mM CaCl The resulting solution was incubated for another 2 hours, and transferred to a 24- well plate. Thereafter, 0.6 mL of DMEM was added thereto, and a concentration of Ca + ions was adjusted to 4.5 mM. After further incubation at 37 0 C for 4 hours, the DNA-bound nanoparticles were washed with a PBS solution. The DNA-bound nanoparticles were added to cells and gene delivery signals were observed.
  • FIG. 8 shows a process using, as a gene delivery system, MNP@SiO
  • FIG. 9 shows confocal laser scanning micrographs of the transfected cells after the gene delivery using the plasmid DNA-bound MNP @ SiO (RITC)-PTMA.
  • 9A is a blue fluorescence micrograph
  • 9B is an optical micrograph
  • 9C is a red fluorescence micrograph
  • 9D is an overlapping micrograph of 9 A, 9B and 9C.
  • Red dots correspond to MNP @ SiO (RITC)-PTMA, and a blue color shows that GFP fluorescence appears in the cytoplasm by DNA transfection.
  • the magnetic nanoparticles according to the present invention can be usefully used as a gene delivery system by binding with the plasmid DNA gene.
  • MNP @ SiO (FITQ-PEG/APS was prep ⁇ ared in the same manner as in Example 1 , except that 3-aminopropyltriethoxysilane (APS) was co-treated upon treatment of magnetic nanoparticles (MNP@SiO (RITC)) with a (CH O) Si-PEG compound in Section 1 of Example 1.
  • PEG/ APS 5/1 (mole ratio), 22.9 mg/mL, amine concentration of 6.5 mmol/g] was added to a solution of maleimidobutyric acid (0.96 g, 1.4 mmol), PyBOP (benzotriazol-l-yl-oxy-tris(pyrrolidino)phosphonium hexafluorophosphate) (0.43 g, 0.826 mmol) and HOBt (N-hydroxybenzotriazole) (0.19 g, 1.4 mmol) in anhydrous DMF.
  • purified diisopropylethylamine 0.2 mL was added to the mixture which was then stirred at room temperature for 20 hours.
  • the reaction materials were transferred to an Eppendorf tube and washed several times with DMF.
  • the nanoparticles were re-dispersed in 0.8 mL of DMF, and stored at room temperature under shielding of light.
  • FIG. 10 shows a process involving co-treatment of (CH O) Si-PEG and APS on the surface of the magnetic nanoparticles, introduction of a maleimide group into an amine group of the surface of the magnetic nanoparticles and introduction of an antibody for recognition of a certain cell thereto.
  • FIG. 11 shows confocal laser scanning micrographs showing utilization of the antibody-bound magnetic nanoparticles in cell staining.
  • FIG. 12 shows confocal laser scanning micrographs showing utilization of the antibody-bound magnetic nanoparticles in cell staining for a leukemia cells and lung cancer cells.
  • Optical micrograph HC: Red fluorescence micrograph
  • HD Overlapping micrograph of 1 IA, 1 IB and 11C.
  • the CD-10 antibody selectively bound to a membrane of a leukemia cell (SP2/O) (12A to 12C), but did not bind to a lung cancer cell (12D to 12F).
  • SP2/O leukemia cell
  • Example 5 Intraperitoneal administration of magnetic nanoparticles according to the present invention into mice ( in vivo experiment)
  • mice were given MNP@SiO (RITC) according to the present invention via intraperitoneal injection and observed at intervals of 15 min via MRI.
  • a control group was not given nanoparticles of the present invention.
  • the magnetic nanoparticles according to the present invention have both optical and magnetic properties and are applicable to various bio-fields.
  • chemical functional groups can be introduced into nano-scale materials, using a variety of compounds.

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Abstract

L'invention concerne des nanoparticules magnétiques (MNP) présentant une fluorescence, ainsi que leur préparation et leur utilisation. Les nanoparticules magnétiques selon l'invention présentent à la fois des propriétés optiques et des propriétés magnétiques et, de ce fait, sont applicables à une variété de bio-champs. Par traitement superficiel des coques de silice des nanoparticules magnétiques par un matériau hydrosoluble, une variété de groupes fonctionnels chimiques peuvent être introduits dans des matériaux à l'échelle nanométrique. De plus, en utilisant des matériaux nanométriques ainsi chimiquement modifiés, il est possible d'augmenter ou de diminuer la pénétrabilité des nanoparticules nanométriques dans des cellules, et il est également possible d'impartir une sélectivité en vue d'agir uniquement sur des cellules spécifiques désirées.
EP06798700A 2005-09-08 2006-09-08 Nanoparticules magnétiques présentant une fluorescence, leur procédé de préparation et leur utilisation Withdrawn EP1934609A4 (fr)

Applications Claiming Priority (3)

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US22096905A 2005-09-08 2005-09-08
KR1020050112245A KR100821192B1 (ko) 2005-09-08 2005-11-23 형광성을 가지는 자성 나노 입자 및 그 제조방법
PCT/KR2006/003569 WO2007029980A1 (fr) 2005-09-08 2006-09-08 Nanoparticules magnetiques presentant une fluorescence, leur procede de preparation et leur utilisation

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EP1934609A1 EP1934609A1 (fr) 2008-06-25
EP1934609A4 true EP1934609A4 (fr) 2009-06-24

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ITFI20080117A1 (it) * 2008-06-26 2009-12-27 Colorobbia Italiana Spa Uso di cobalto ferriti come agenti di contrasto per risonanza magnetica.
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