WO2019127836A1 - Magnetic microsphere and preparation method and application thereof - Google Patents

Magnetic microsphere and preparation method and application thereof Download PDF

Info

Publication number
WO2019127836A1
WO2019127836A1 PCT/CN2018/074903 CN2018074903W WO2019127836A1 WO 2019127836 A1 WO2019127836 A1 WO 2019127836A1 CN 2018074903 W CN2018074903 W CN 2018074903W WO 2019127836 A1 WO2019127836 A1 WO 2019127836A1
Authority
WO
WIPO (PCT)
Prior art keywords
polymer
magnetic
microsphere
polymerization
microspheres
Prior art date
Application number
PCT/CN2018/074903
Other languages
French (fr)
Chinese (zh)
Inventor
江必旺
赵光耀
Original Assignee
苏州纳微科技股份有限公司
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
Application filed by 苏州纳微科技股份有限公司 filed Critical 苏州纳微科技股份有限公司
Publication of WO2019127836A1 publication Critical patent/WO2019127836A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F285/00—Macromolecular compounds obtained by polymerising monomers on to preformed graft polymers
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F257/00—Macromolecular compounds obtained by polymerising monomers on to polymers of aromatic monomers as defined in group C08F12/00
    • C08F257/02—Macromolecular compounds obtained by polymerising monomers on to polymers of aromatic monomers as defined in group C08F12/00 on to polymers of styrene or alkyl-substituted styrenes
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F8/00—Chemical modification by after-treatment
    • C08F8/18—Introducing halogen atoms or halogen-containing groups
    • C08F8/24—Haloalkylation
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F8/00—Chemical modification by after-treatment
    • C08F8/30—Introducing nitrogen atoms or nitrogen-containing groups
    • C08F8/32—Introducing nitrogen atoms or nitrogen-containing groups by reaction with amines
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
    • C08K3/00—Use of inorganic substances as compounding ingredients
    • C08K3/18—Oxygen-containing compounds, e.g. metal carbonyls
    • C08K3/20—Oxides; Hydroxides
    • C08K3/22—Oxides; Hydroxides of metals
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • 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
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08K—Use of inorganic or non-macromolecular organic substances as compounding ingredients
    • C08K3/00—Use of inorganic substances as compounding ingredients
    • C08K3/18—Oxygen-containing compounds, e.g. metal carbonyls
    • C08K3/20—Oxides; Hydroxides
    • C08K3/22—Oxides; Hydroxides of metals
    • C08K2003/2265—Oxides; Hydroxides of metals of iron

Definitions

  • the present disclosure belongs to the technical field of polymer synthesis, and relates to a magnetic microsphere and a preparation method and application thereof.
  • Magnetic microspheres generally refer to composites of magnetic materials with other non-magnetic inorganic or polymeric matrices and are spherical particles having submicron to micron sizes. Magnetic microspheres have a wide range of applications in many fields, particularly bioassays and in vitro diagnostic reagents. Magnetic microspheres for diagnostic reagents require uniform particle size, fast magnetic response, good suspension and low non-specific adsorption to biological macromolecules such as proteins and nucleic acids.
  • the first is to prepare the core of the solid magnetic material, and then to coat the functionalized outer layer to obtain the core-shell structure magnetic microspheres; the second is to use non-magnetic porous or non-porous micro-
  • the ball is a template for depositing magnetic material, and then the functionalized outer layer is coated to obtain a non-magnetic-magnetic-nonmagnetic "sandwich" type sandwich structure; the third is to add small-sized magnetic nanoparticles during the preparation of the polymer microsphere.
  • a structure in which magnetic nanoparticles are uniformly distributed throughout the entire microsphere is obtained.
  • EP 0 106 973 B1 discloses a process for preparing a porous polymer microsphere template by seed emulsion polymerization and then depositing a magnetic iron oxide material in the pores to prepare magnetic microspheres.
  • the porous polymer microspheres used in this method have a specific surface area of 100-200 m 2 /g, and the microspheres after deposition of the magnetic material still have a pore structure, and these pores are easily non-specific to biomacromolecules such as proteins in subsequent bioassay experiments. Adsorption, resulting in higher background noise.
  • JP2004-205481 discloses a method of preparing magnetic microspheres by adding small particle size magnetic nanoparticles during emulsion polymerization.
  • the method can directly obtain non-porous magnetic microspheres to avoid non-specific adsorption caused by the pore structure, but the addition of magnetic nanoparticles during the polymerization process easily affects the nucleation process of the polymer, and it is difficult to obtain monodisperse magnetic microparticles of uniform size. ball.
  • the structure of the magnetic microsphere itself determines its performance in bioassay applications.
  • the magnetic microspheres with uniform synthetic size, high magnetic material content and no pore structure are the basis of a diagnostic kit with stable development batch, low background noise and high sensitivity.
  • the present disclosure provides a magnetic microsphere and a preparation method and application thereof.
  • the present disclosure provides a magnetic microsphere comprising a polymer core, a brush-like polymer interlayer with a magnetic material, and a functionalized polymer outer layer, the brush-like polymer Functional group.
  • the magnetic microsphere provided by the present disclosure is a magnetic microsphere of a "sandwich" type sandwich structure, and the brush-like polymer of the intermediate layer of the magnetic microsphere provided by the present disclosure has a functional group, and the functional group serves as a binding site of the metal oxide. It can effectively increase the deposition amount of magnetic materials and solve the problem of small magnetic material content in magnetic microspheres in the past.
  • the polymer core has a diameter of from 0.01 ⁇ m to 1000 ⁇ m, such as 0.01 ⁇ m, 1 ⁇ m, 10 ⁇ m, 50 ⁇ m, 100 ⁇ m, 500 ⁇ m, 1000 ⁇ m, and the like.
  • the polymer core is obtained by polymerization of a monomer having an unsaturated bond.
  • the polymer core is obtained by polymerization of an aromatic monomer containing an unsaturated bond.
  • the polymer core is a polystyrene microsphere.
  • the polymer core is obtained by polymerization of an acrylate or methacrylate monomer containing an unsaturated bond.
  • the brush-like polymer is obtained by polymerization from an acrylate or methacrylate monomer containing an unsaturated bond.
  • the brush polymer is any one of polyglycidyl methacrylate, poly-tert-butyl methacrylate or poly-2,2-dihydroxypropyl 2-methyl-2-acrylate. A combination of at least two.
  • the brush-like polymer forms a high-density polymer chain on the surface of the polymer core by surface-initiated polymerization, since the distance between the polymer chains is less than the free radius of gyration of the polymer chain, the polymer chain The configuration is limited and is almost perpendicular to the surface of the core microspheres to form a brush-like structure.
  • the brush-like polymer carries a functional group of any one of a hydroxyl group, an amino group, a carboxyl group, a thiol group or an epoxy group or a combination of at least two.
  • the magnetic material is any one or a combination of at least two of Fe 3 O 4 , ⁇ -Fe 2 O 3 or MFe 2 O 4 , and M is a metal other than Fe.
  • M in the MFe 2 O 4 is any one of Co, Mg, Ni, Mn or Zn or a combination of at least two.
  • the functionalized polymer outer layer comprises at least one layer of functionalized polymer, which may be a layer of functionalized polymer, or may be two layers, three layers, or the like.
  • the functionalized polymer is obtained by polymerization from a monomer containing an unsaturated bond.
  • the unsaturated bond-containing monomer is any one of styrene, methacrylic acid, 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester or glycidyl methacrylate or A combination of at least two.
  • the functional group of the functionalized polymer is any one or a combination of at least two of a hydroxyl group, an amino group, a carboxyl group, a thiol group, an aldehyde group, an epoxy group or a p-toluenesulfonyl group.
  • the magnetic microspheres provided by the present disclosure use non-porous polymer microspheres instead of porous polymer microspheres as a template (core) of magnetic microspheres, which avoids the pores in the microspheres may cause proteins and other organisms in subsequent bioassay experiments. Non-specific adsorption of macromolecules and high background noise.
  • the present disclosure provides a method of preparing a magnetic microsphere as described above, the method comprising the steps of:
  • the surface of the magnetic material-containing microsphere obtained in the step (3) is polymerized to form an outer layer of the functionalized polymer to obtain the magnetic microsphere.
  • a high-density functional group can be introduced, and a magnetic material can be deposited as a template to prepare a magnetic microsphere having a high magnetic material content.
  • the magnetic material can be prepared by a coprecipitation method: a soluble metal salt is added to the microsphere dispersion in the presence of a microsphere of a brush-like polymer template having a functional group on the surface, and the metal cation is bonded to the functional group of the brush polymer. Then, an alkali solution is added to the dispersion to adjust the pH, and the metal cation is precipitated from the solution to form small-sized metal oxide small particles.
  • the preparation method provided by the present disclosure is simple and easy, and the magnetic microspheres prepared by the preparation method provided by the present disclosure are uniform in size and have no pore structure. Magnetic microspheres with uniform dimensions and no pore structure are the basis for the development of diagnostic kits with good stability, low background noise and high sensitivity.
  • the preparation method further includes preparing the polymer core by a polymerization reaction.
  • the polymerization reaction is any one or a combination of at least two of emulsion polymerization, microemulsion polymerization, miniemulsion polymerization, soap-free emulsion polymerization, dispersion polymerization, suspension polymerization or seed polymerization.
  • the technical solution of the present disclosure first synthesizes the polymer core microspheres and then introduces the magnetic material, thereby avoiding the problem that the microspheres are not uniform in size by adding the magnetic materials while synthesizing the polymer microspheres, and the magnetic microspheres having a uniform height can be obtained. Better batch stability in subsequent bioassay applications.
  • the brush-like polymer intermediate layer of the magnetic microsphere provided by the present disclosure provides a large number of functional groups as a binding site of the metal oxide, can effectively increase the deposition amount of the magnetic material, and compensates for the non-porous polymer microsphere as a template.
  • the magnetic material content of the magnetic microspheres is generally less than that of the magnetic material content of the magnetic microspheres using the porous polymer microspheres as a template.
  • the polymer core may be a polystyrene microsphere, and the preparation method is as follows:
  • the initiator is added to the solvent obtained in the step I, stirred and dissolved, added with an emulsifier, and mixed;
  • polystyrene seeds are added to the emulsion obtained in the step II, and reacted to obtain polystyrene microspheres.
  • the crosslinking agent in step I is divinylbenzene.
  • the solvent of step I is cyclohexanol.
  • the mass ratio of the styrene, the crosslinking agent and the solvent in the step I is (25-35):(50-60):(100-120), for example 25:50:100, 30:55:110, 35:60:120, 30:60:100, and the like.
  • the initiator in step II is azobisisobutyronitrile, and the initiator is used in an amount of 3-5 wt% of styrene, such as 3 wt%, 4 wt%, 5 wt%, and the like.
  • the emulsifier of step II is sodium octadecyl sulfonate.
  • the polystyrene seed in the step III is a polystyrene microsphere having a particle diameter of 0.005 to 1000 ⁇ m, and the 0.005 to 1000 ⁇ m may be 0.005 ⁇ m, 0.01 ⁇ m, 0.1 ⁇ m, 1 ⁇ m, 10 ⁇ m, 100 ⁇ m, 1000 ⁇ m or the like.
  • the reaction in the step III is a reaction at room temperature for 24 hours and then the temperature is raised to 80-100 ° C.
  • the 80-100 ° C may be 80 ° C, 85 ° C, 90 ° C, 95 ° C, 100 ° C, and the like.
  • the polystyrene microspheres obtained in the step III have a particle diameter of 0.01 ⁇ m to 1000 ⁇ m, for example, 0.01 ⁇ m, 0.1 ⁇ m, 1 ⁇ m, 10 ⁇ m, 100 ⁇ m, 1000 ⁇ m, or the like.
  • the preparation method of the magnetic microspheres is as follows:
  • reaction solution B (2) adding the polymer core to the solvent, ultrasonication, to obtain a reaction solution B;
  • microspheres obtained in the step (5) are dispersed in a solvent, a polymer monomer and an initiator are added, and polymerization is carried out to form an outer layer of the functionalized polymer to obtain magnetic microspheres.
  • the brush-like polymer monomer in the step (1) is any one of glycidyl methacrylate, t-butyl methacrylate or 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester.
  • the solvent in the step (1) is isopropyl alcohol, and the amount of the brush-like polymer monomer is 1 mol/L (solution).
  • the catalyst of the step (1) comprises cuprous chloride, cupric chloride and tris(2-dimethylaminoethyl)amine, the cuprous chloride, cupric chloride and tris(2-dimethylamino).
  • the mass ratio of ethylamine addition is (5-10):(1-2):(10-20), for example 5:1:10, 8:2:15, 10:2:20, 7:1: 20 and so on.
  • step (1) is to add a catalyst under a nitrogen atmosphere and stir.
  • step (1) further comprises deoxidizing the solution to nitrogen for 1-2 hours, for example 1 hour, 1.5 hours, 2 hours, etc., prior to the addition of the catalyst.
  • the solvent in the step (2) is isopropyl alcohol.
  • the polymer core of the step (2) is added in an amount of 50-60 g/L (solvent), for example, 50 g/L (solvent), 52 g/L (solvent), 55 g/L (solvent), 57 g/L. (solvent), 60 g/L (solvent), and the like.
  • solvent for example, 50 g/L (solvent), 52 g/L (solvent), 55 g/L (solvent), 57 g/L. (solvent), 60 g/L (solvent), and the like.
  • step (2) further comprises deoxygenating with nitrogen for 1-2 hours after sonication, for example 1 hour, 1.5 hours, 2 hours, and the like.
  • the reaction time in the step (3) is 24-30 h, such as 24 h, 26 h, 28 h, 30 h, and the like.
  • step (3) further comprises introducing air to stop the reaction.
  • the step (3) further comprises, after the reaction is completed, filtering, washing and drying the reaction solution.
  • the microspheres in the step (4) are added in an amount of 50-60 g/L (solvent), for example, 50 g/L (solvent), 52 g/L (solvent), 55 g/L (solvent), 57 g/L ( Solvent), 60 g/L (solvent), and the like.
  • solvent for example, 50 g/L (solvent), 52 g/L (solvent), 55 g/L (solvent), 57 g/L ( Solvent), 60 g/L (solvent), and the like.
  • the functional monomer in the step (4) is ethylenediamine and/or hydrochloric acid.
  • the solvent in the step (4) is anhydrous ethanol and/or deionized water.
  • the reaction in the step (4) is a reaction at room temperature for 6 to 24 hours, for example, 6 hours, 10 hours, 15 hours, 20 hours, 24 hours, and the like.
  • the step (4) further comprises filtering, washing and drying the reaction solution after the reaction is completed.
  • the solvent in the step (5) is deionized water.
  • the functional group-containing microspheres are added in an amount of 15-25 g/L (solvent), for example, 15 g/L (solvent), 17 g/L (solvent), 20 g/L (solvent). ), 22 g / L (solvent), 25 g / L (solvent) and the like.
  • the method of depositing the magnetic material in the step (5) is to add ferric chloride hexahydrate and/or ferrous chloride tetrahydrate to the microsphere solution at 0-5 ° C, stir under nitrogen, and then Ammonia water was added dropwise, and after completion of the dropwise addition, the mixture was stirred at room temperature overnight, and the 0-5 ° C may be 0 ° C, 1 ° C, 2 ° C, 3 ° C, 4 ° C, 5 ° C, and the like.
  • the amount of the magnetic material added in the step (5) is 2-4 times, that is, 2 times, 2.5 times, 3 times, 3.5 times, 4 times, etc., of the microspheres added in the step (5).
  • step (5) further comprises removing the solvent after the surface deposition of the magnetic material, washing, washing and redispersing in deionized water.
  • the solvent in the step (6) is a mixture of deionized water and a Brij-35 (30%) aqueous solution.
  • the polymer monomer in the step (6) is any one of styrene, methacrylic acid, glycidyl methacrylate or 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester. Or a combination of at least two.
  • the initiator in the step (6) is tert-butyl (2-ethylhexanoate) peroxide.
  • the polymerization in the step (6) is carried out under a nitrogen atmosphere.
  • the reaction temperature of the polymerization reaction in the step (6) is 80 to 100 ° C, for example, 80 ° C, 85 ° C, 90 ° C, 95 ° C, 100 ° C, and the like.
  • the reaction time of the polymerization reaction in the step (6) is 8 to 10 hours, for example, 8 hours, 8.5 hours, 9 hours, 9.5 hours, 10 hours, and the like.
  • step (6) further comprises washing the reaction solution after the end of the polymerization reaction, washing and redispersing in deionized water.
  • the present disclosure provides the use of a magnetic microsphere as described above in the preparation of a diagnostic kit.
  • Uniform magnetic microspheres have better batch stability in subsequent bio-detection applications.
  • the magnetic microspheres provided by the present disclosure have uniform size, high magnetic material content and no pore structure, and are stable in development and background noise. The basis for a low-sensitivity diagnostic kit.
  • the magnetic microsphere provided by the present disclosure is a magnetic microsphere of a "sandwich" type sandwich structure, and the brush-like polymer of the intermediate layer of the magnetic microsphere provided by the present disclosure has a functional group, and the functional group serves as a binding site of the metal oxide. It can effectively increase the deposition amount of magnetic materials and solve the problem of small magnetic material content in magnetic microspheres in the past.
  • the preparation method provided by the present disclosure is simple and easy.
  • the polymer core used in Example 1-2 was a 5 ⁇ m polystyrene microsphere, manufactured by Suzhou Nawei Technology Co., Ltd., model No. UniPS-NP5.
  • the magnetic microspheres provided in this embodiment include a polystyrene microsphere core having a diameter of 5 ⁇ m, an intermediate layer of brush-like glycidyl methacrylate with magnetic iron oxide, and an outer layer of a functionalized polymer, brush-like methacrylic acid.
  • Glycidyl esters carry an amino group.
  • the preparation method is as follows:
  • the solution was removed by filtration through a sand core funnel, and then the microspheres were washed with 200 mL of the following solvent: deionized water, 5% hydrochloric acid, deionized water, tetrahydrofuran, ethanol, acetone.
  • the washed microspheres were dried at 60 ° C for 12 hours to obtain chloromethylated microspheres.
  • the monomer glyceryl methacrylate was dissolved in isopropyl alcohol to prepare a solution having a concentration of 1 mol/L.
  • 100 mL of the above-mentioned monomeric isopropanol solution was placed in a 250 mL three-necked flask, deoxygenated with nitrogen at 25 ° C for 1 hour, and then 115 mg of cuprous chloride, 15.7 mg of copper chloride and 300 mg of tris(2) were sequentially added under nitrogen protection.
  • -Dimethylaminoethyl)amine mechanically stirred for 15 minutes to obtain a reaction solution A1.
  • reaction solution B1 Another dry 250 mL three-necked flask was taken, and 1.0 g of the microspheres obtained in the step (1) and 20 mL of isopropyl alcohol were added, and ultrasonically dispersed, and deoxidized by nitrogen gas at 25 ° C for 1 hour to obtain a reaction solution B1.
  • reaction solution A1 was added to the reaction solution B1 under a nitrogen atmosphere to initiate polymerization. After mechanical stirring at 25 ° C for 24 hours, air was bubbled through the solution to terminate the reaction. The solution was removed by filtration using a sand core funnel, and then the microspheres were washed with 50 mL of the following solvent: methanol, 50 mM aqueous EDTA (pH 8.0), deionized water (twice), and acetone. The washed microspheres were dried at 60 ° C for 12 hours to obtain polymer microspheres of surface-grafted brush-like polyglycidyl methacrylate.
  • microspheres obtained in the step (2) was ultrasonically dispersed in 20 mL of absolute ethanol, and 5 mL of anhydrous ethylenediamine was added thereto, and the reaction was rotated at room temperature for 6 hours.
  • the solution was removed by filtration through a sand funnel, washed twice with absolute ethanol, 50 mL each time, and then washed twice with deionized water, 50 mL each time, and finally washed once with 50 mL of acetone.
  • the washed microspheres were dried at 60 ° C for 12 hours to obtain microspheres having amino groups on the surface.
  • 1.0 g of the microspheres obtained in the step (3) was placed in a 250 mL three-necked flask, and 50 mL of deionized water was added to the flask to ultrasonically disperse the microspheres.
  • the flask was immersed in an ice water bath, and the temperature of the solution was lowered to below 5 ° C.
  • 2.0 g of iron (III) chloride hexahydrate and 0.93 g of ferrous chloride tetrahydrate were added to the flask.
  • Stirring was continued for 1 hour under nitrogen, and then 8 mL of aqueous ammonia was slowly added to the solution using a dropping funnel.
  • micro-magnetic iron oxide After the addition was completed, the ice water bath was removed and stirring was continued at room temperature overnight. After the reaction is completed, the magnetic iron oxide-deposited microspheres are separated by a magnet, the supernatant is removed, and then washed three times with 50 mL of deionized water, and finally dispersed in 25 mL of deionized water to obtain micro-magnetic iron oxide. ball. The microsphere dispersion was stored at 2-8 °C.
  • 1.0 g of the microspheres obtained in the step (4) was placed in a 100 mL three-necked flask, and 30 mL of deionized water and 0.5 mL of Brij-35 30% aqueous solution were added to ultrasonically disperse the microspheres.
  • To the above dispersion was added 0.2 g of styrene, deoxygenated with nitrogen at 25 ° C for 1 hour, then 0.05 g of initiator (2-ethylhexanoic acid) tert-butyl ester was added, and the temperature was raised to 80 ° C under nitrogen atmosphere. Initiate polymerization.
  • the reaction system was cooled to room temperature to obtain a sandwich-structure magnetic microsphere having a surface coated with a polymer.
  • the product was washed three times with ethanol, 50 mL each time, then ultrasonically dispersed in 30 mL of an aqueous solution containing 0.5% Brij-35, and 0.2 g of styrene, 0.01 g of methacrylic acid and 0.02 g of glycidol methacrylate were added to the dispersion.
  • the ester was deoxygenated with nitrogen at 25 ° C for 1 hour, then 0.02 g of initiator (2-ethylhexanoic acid) tert-butyl ester was added, and the temperature was raised to 80 ° C under nitrogen atmosphere to initiate polymerization. After reacting for 8 hours, the reaction system was cooled to room temperature to obtain sandwich-structured magnetic microspheres having a surface coated with two layers of polymer. The product was washed twice with 50 mL each time, once with 50 mL of deionized water, and finally in 25 mL of deionized water and stored at 2-8 °C.
  • the magnetic microsphere provided in this embodiment comprises a core of polystyrene microspheres having a diameter of 5 ⁇ m, an intermediate layer of brush-like tert-butyl methacrylate with magnetic iron oxide, and an outer layer of functionalized polymer, brush-like methacrylic acid.
  • Tert-butyl ester has a carboxyl group.
  • the preparation method is as follows:
  • the monomer tert-butyl methacrylate was dissolved in isopropyl alcohol to prepare a solution having a concentration of 1 mol/L.
  • 100 mL of the above-mentioned monomeric isopropanol solution was placed in a 250 mL three-necked flask, deoxygenated with nitrogen at 25 ° C for 1 hour, and then 115 mg of cuprous chloride, 15.7 mg of copper chloride and 300 mg of tri 2-Dimethylaminoethyl)amine was mechanically stirred for 15 minutes to obtain a reaction solution A2.
  • reaction solution A2 was added to the reaction solution B2 under a nitrogen atmosphere to initiate polymerization. After mechanical stirring at 25 ° C for 24 hours, air was bubbled through the solution to terminate the reaction. The solution was removed by filtration using a sand core funnel, and then the microspheres were washed with 50 mL of the following solvent: methanol, 50 mM aqueous EDTA (pH 8.0), deionized water (twice), and acetone. The washed microspheres were dried at 60 ° C for 12 hours to obtain polymer microspheres of surface-grafted brush-like polybutyl methacrylate.
  • 1.0 g of the microspheres obtained in the step (2) was ultrasonically dispersed in 20 mL of deionized water, and the pH was adjusted to 1 with 6 mol/L hydrochloric acid. The reaction was rotated overnight at room temperature. The solution was removed by filtration through a sand core funnel, washed twice with 0.1 mol/L sodium hydroxide solution, 50 mL each time, and then washed with deionized water, 50 mL each time until the filtrate pH was neutral, and finally washed once with 50 mL of acetone. The washed microspheres were dried at 60 ° C for 12 hours to obtain carboxyl group-containing microspheres.
  • 1.0 g of the microspheres obtained in the step (3) was placed in a 250 mL three-necked flask, and 50 mL of deionized water was added to the flask to ultrasonically disperse the microspheres.
  • the flask was immersed in an ice water bath, and the temperature of the solution was lowered to below 5 ° C.
  • 2.0 g of iron (III) chloride hexahydrate and 0.93 g of ferrous chloride tetrahydrate were added to the flask.
  • Stirring was continued for 1 hour under nitrogen, and then 8 mL of aqueous ammonia was slowly added to the solution using a dropping funnel.
  • micro-magnetic iron oxide After the addition was completed, the ice water bath was removed and stirring was continued at room temperature overnight. After the reaction is completed, the magnetic iron oxide-deposited microspheres are separated by a magnet, the supernatant is removed, and then washed three times with 50 mL of deionized water, and finally dispersed in 25 mL of deionized water to obtain micro-magnetic iron oxide. ball. The microsphere dispersion was stored at 2-8 °C.
  • 1.0 g of the microspheres obtained in the step (4) was placed in a 100 mL three-necked flask, and 30 mL of deionized water and 0.5 mL of Brij-35 30% aqueous solution were added to ultrasonically disperse the microspheres.
  • 0.2 g of styrene and 0.01 g of methacrylic acid were added, and deoxygenated with nitrogen at 25 ° C for 1 hour, and then 0.02 g of an initiator (2-ethylhexanoic acid) tert-butyl ester was added thereto, under nitrogen.
  • the temperature was raised to 80 ° C under protection to initiate polymerization.
  • reaction system After reacting for 8 hours, the reaction system was cooled to room temperature to obtain magnetic microspheres having a surface coated with a layer of a polymer.
  • the product was washed twice with 50 mL each time, once with 50 mL of deionized water, and finally in 25 mL of deionized water and stored at 2-8 °C.
  • the magnetic microspheres provided in this embodiment include a polystyrene microsphere core having a diameter of 5 ⁇ m, an intermediate layer and a function of brush-like 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester with magnetic iron oxide.
  • the outer layer of the polymer is brushed with 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester with a hydroxyl group.
  • a solution of 2 mol/L was prepared by dissolving 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester in isopropanol.
  • 100 mL of the above-mentioned monomeric isopropanol solution was placed in a 250 mL three-necked flask, deoxygenated with nitrogen at 25 ° C for 1 hour, and then 115 mg of cuprous chloride, 15.7 mg of copper chloride and 300 mg of tris(2) were sequentially added under nitrogen protection.
  • -Dimethylaminoethyl)amine mechanically stirred for 15 minutes to obtain a reaction solution A3.
  • reaction solution A3 was added to the reaction solution B3 under a nitrogen atmosphere to initiate polymerization. After mechanical stirring at 25 ° C for 24 hours, air was bubbled through the solution to terminate the reaction.
  • the solution was removed by filtration using a sand core funnel, and then the microspheres were washed with 50 mL of the following solvent: methanol, 50 mM aqueous EDTA (pH 8.0), deionized water (twice), and acetone.
  • the washed microspheres were dried at 60 ° C for 12 hours to obtain polymer microspheres having a surface grafted brush-like poly(2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester).
  • 1.0 g of the microspheres obtained in the step (3) was placed in a 250 mL three-necked flask, and 50 mL of deionized water was added to the flask to ultrasonically disperse the microspheres.
  • the flask was immersed in an ice water bath, and the temperature of the solution was lowered to below 5 ° C.
  • 2.0 g of iron (III) chloride hexahydrate and 0.93 g of ferrous chloride tetrahydrate were added to the flask.
  • Stirring was continued for 1 hour under nitrogen, and then 8 mL of aqueous ammonia was slowly added to the solution using a dropping funnel.
  • micro-magnetic iron oxide After the addition was completed, the ice water bath was removed and stirring was continued at room temperature overnight. After the reaction is completed, the magnetic iron oxide-deposited microspheres are separated by a magnet, the supernatant is removed, and then washed three times with 50 mL of deionized water, and finally dispersed in 25 mL of deionized water to obtain micro-magnetic iron oxide. ball. The microsphere dispersion was stored at 2-8 °C.
  • 1.0 g of the microspheres obtained in the step (4) was placed in a 100 mL three-necked flask, and 30 mL of deionized water and 0.5 mL of Brij-35 30% aqueous solution were added to ultrasonically disperse the microspheres.
  • 0.2 g of styrene, 0.01 g of methacrylic acid and 0.02 g of 2,3-dihydroxypropyl 2-methyl-2-acrylate were added to the above dispersion, and deoxygenated under nitrogen at 25 ° C for 1 hour, and then added.
  • 0.02 g of initiator, tert-butyl peroxy (2-ethylhexanoate) was heated to 80 ° C under nitrogen to initiate polymerization.
  • reaction system After reacting for 8 hours, the reaction system was cooled to room temperature to obtain magnetic microspheres having a surface coated with a layer of a polymer.
  • the product was washed twice with 50 mL each time, once with 50 mL of deionized water, and finally in 25 mL of deionized water and stored at 2-8 °C.

Abstract

A magnetic microsphere with a sandwich structure and a preparation method thereof. The magnetic microsphere comprises a polymer core, a brush polymer intermediate layer with a magnetic material, and a functionalized polymer outer layer. A functional group of the brush polymer acts as a binding site of a metal oxide, thereby increasing the deposition amount of the magnetic material.

Description

一种磁性微球及其制备方法和应用Magnetic microsphere and preparation method and application thereof 技术领域Technical field
本公开属于聚合物合成技术领域，涉及一种磁性微球及其制备方法和应用。The present disclosure belongs to the technical field of polymer synthesis, and relates to a magnetic microsphere and a preparation method and application thereof.
背景技术Background technique
磁性微球通常指磁性材料与其他非磁性无机或聚合物基质复合而成的，是具有亚微米至微米尺寸的球形颗粒。磁性微球在许多领域特别是生物检测与体外诊断试剂中具有广泛的应用。用于诊断试剂的磁性微球要求其粒径均一、磁响应快、悬浮性好并且对蛋白、核酸等生物大分子具有较低的非特异性吸附。磁性微球的常见制备方式有三种：第一种是先制备实心磁性材料内核，再包被功能化外层得到核-壳结构磁性微球；第二种是以非磁性的多孔或者无孔微球为模板沉积磁性材料，再包被功能化外层得到非磁性-磁性-非磁性的“三明治”型夹心结构；第三种是在制备聚合物微球过程中加入小粒径磁性纳米粒子，得到磁性纳米粒子均匀分布在整个微球内部的结构。Magnetic microspheres generally refer to composites of magnetic materials with other non-magnetic inorganic or polymeric matrices and are spherical particles having submicron to micron sizes. Magnetic microspheres have a wide range of applications in many fields, particularly bioassays and in vitro diagnostic reagents. Magnetic microspheres for diagnostic reagents require uniform particle size, fast magnetic response, good suspension and low non-specific adsorption to biological macromolecules such as proteins and nucleic acids. There are three common preparation methods for magnetic microspheres: the first is to prepare the core of the solid magnetic material, and then to coat the functionalized outer layer to obtain the core-shell structure magnetic microspheres; the second is to use non-magnetic porous or non-porous micro- The ball is a template for depositing magnetic material, and then the functionalized outer layer is coated to obtain a non-magnetic-magnetic-nonmagnetic "sandwich" type sandwich structure; the third is to add small-sized magnetic nanoparticles during the preparation of the polymer microsphere. A structure in which magnetic nanoparticles are uniformly distributed throughout the entire microsphere is obtained.
EP0106973B1公开了一种以种子乳液聚合制备多孔聚合物微球模板，然后在孔道内沉积磁性氧化铁材料制备磁性微球的方法。此法所用的多孔聚合物微球比表面积在100-200m 2/g，沉积磁性材料后的微球仍然具有孔道结构，这些孔道在后续的生物检测实验中容易对蛋白等生物大分子产生非特异性吸附，造成较高的背景噪声。 EP 0 106 973 B1 discloses a process for preparing a porous polymer microsphere template by seed emulsion polymerization and then depositing a magnetic iron oxide material in the pores to prepare magnetic microspheres. The porous polymer microspheres used in this method have a specific surface area of 100-200 m 2 /g, and the microspheres after deposition of the magnetic material still have a pore structure, and these pores are easily non-specific to biomacromolecules such as proteins in subsequent bioassay experiments. Adsorption, resulting in higher background noise.
JP2004-205481公开了一种在乳液聚合过程中加入小粒径磁性纳米粒子来制备磁性微球的方法。此方法可直接得到无孔的磁性微球，避免由孔道结构导致的非特异性吸附，但在聚合过程中加入磁性纳米粒子容易影响聚合物的成核过程，难以制得尺寸均一的单分散磁性微球。JP2004-205481 discloses a method of preparing magnetic microspheres by adding small particle size magnetic nanoparticles during emulsion polymerization. The method can directly obtain non-porous magnetic microspheres to avoid non-specific adsorption caused by the pore structure, but the addition of magnetic nanoparticles during the polymerization process easily affects the nucleation process of the polymer, and it is difficult to obtain monodisperse magnetic microparticles of uniform size. ball.
磁性微球本身的结构决定了其在生物检测应用中的性能。合成尺寸均一、磁性材料含量高且不具有孔道结构的磁性微球是开发批次稳定好、背景噪声低、灵敏度高的诊断试剂盒的基础。The structure of the magnetic microsphere itself determines its performance in bioassay applications. The magnetic microspheres with uniform synthetic size, high magnetic material content and no pore structure are the basis of a diagnostic kit with stable development batch, low background noise and high sensitivity.
发明内容Summary of the invention
以下是对本文详细描述的主题的概述。本概述并非是为了限制权利要求的保护范围。The following is an overview of the topics detailed in this document. This Summary is not intended to limit the scope of the claims.
本公开提供了一种磁性微球及其制备方法和应用。The present disclosure provides a magnetic microsphere and a preparation method and application thereof.
为达此目的，本公开采用以下技术方案：To this end, the present disclosure adopts the following technical solutions:
一方面，本公开提供了一种磁性微球，所述磁性微球包括聚合物内核、带有磁性材料的刷状聚合物中间层和功能化聚合物外层，所述刷状聚合物带有功能基团。In one aspect, the present disclosure provides a magnetic microsphere comprising a polymer core, a brush-like polymer interlayer with a magnetic material, and a functionalized polymer outer layer, the brush-like polymer Functional group.
本公开提供的磁性微球为“三明治”型夹心结构的磁性微球，本公开提供的磁性微球中间层的刷状聚合物带有功能基团，功能基团作为金属氧化物的结合位点，能有效提高磁性材料的沉积量，解决了以往磁性微球中磁性材料含量小的问题。The magnetic microsphere provided by the present disclosure is a magnetic microsphere of a "sandwich" type sandwich structure, and the brush-like polymer of the intermediate layer of the magnetic microsphere provided by the present disclosure has a functional group, and the functional group serves as a binding site of the metal oxide. It can effectively increase the deposition amount of magnetic materials and solve the problem of small magnetic material content in magnetic microspheres in the past.
在本公开中，所述聚合物内核的直径为0.01μm-1000μm，例如0.01μm、1μm、10μm、50μm、100μm、500μm、1000μm等。In the present disclosure, the polymer core has a diameter of from 0.01 μm to 1000 μm, such as 0.01 μm, 1 μm, 10 μm, 50 μm, 100 μm, 500 μm, 1000 μm, and the like.
优选地，所述聚合物内核由带有不饱和键的单体通过聚合得到。Preferably, the polymer core is obtained by polymerization of a monomer having an unsaturated bond.
在本公开中，所述聚合物内核由含有不饱和键的芳香族单体通过聚合得到。In the present disclosure, the polymer core is obtained by polymerization of an aromatic monomer containing an unsaturated bond.
优选地，所述聚合物内核为聚苯乙烯微球。Preferably, the polymer core is a polystyrene microsphere.
优选地，所述聚合物内核由含有不饱和键的丙烯酸酯或甲基丙烯酸酯类单体通过聚合得到。Preferably, the polymer core is obtained by polymerization of an acrylate or methacrylate monomer containing an unsaturated bond.
在本公开中，所述刷状聚合物由含有不饱和键的丙烯酸酯或甲基丙烯酸酯 类单体通过聚合得到。In the present disclosure, the brush-like polymer is obtained by polymerization from an acrylate or methacrylate monomer containing an unsaturated bond.
优选地，所述刷状聚合物为聚甲基丙烯酸缩水甘油酯、聚甲基丙烯酸叔丁酯或聚2-甲基-2-丙烯酸-2，3-二羟基丙酯中的任意一种或至少两种的组合。Preferably, the brush polymer is any one of polyglycidyl methacrylate, poly-tert-butyl methacrylate or poly-2,2-dihydroxypropyl 2-methyl-2-acrylate. A combination of at least two.
在本公开中，所述刷状聚合物是通过表面引发聚合反应在聚合物内核表面形成高密度聚合物链，由于聚合物链之间的距离小于聚合物链的自由回转半径，聚合物链的构型受限，与内核微球的表面几乎垂直，形成刷状结构。In the present disclosure, the brush-like polymer forms a high-density polymer chain on the surface of the polymer core by surface-initiated polymerization, since the distance between the polymer chains is less than the free radius of gyration of the polymer chain, the polymer chain The configuration is limited and is almost perpendicular to the surface of the core microspheres to form a brush-like structure.
优选地，所述刷状聚合物带有的功能基团为羟基、氨基、羧基、巯基或环氧基中的任意一种或至少两种的组合。Preferably, the brush-like polymer carries a functional group of any one of a hydroxyl group, an amino group, a carboxyl group, a thiol group or an epoxy group or a combination of at least two.
在本公开中，所述磁性材料为Fe 3O 4、γ-Fe 2O 3或MFe 2O 4中的任意一种或至少两种的组合，M为除Fe以外的金属。 In the present disclosure, the magnetic material is any one or a combination of at least two of Fe 3 O 4 , γ-Fe 2 O 3 or MFe 2 O 4 , and M is a metal other than Fe.
优选地，所述MFe 2O 4中的M为Co、Mg、Ni、Mn或Zn中的任意一种或至少两种的组合。 Preferably, M in the MFe 2 O 4 is any one of Co, Mg, Ni, Mn or Zn or a combination of at least two.
在本公开中，所述功能化聚合物外层包括至少一层功能化聚合物，可以是一层功能化聚合物，也可以是两层、三层等。In the present disclosure, the functionalized polymer outer layer comprises at least one layer of functionalized polymer, which may be a layer of functionalized polymer, or may be two layers, three layers, or the like.
优选地，所述功能化聚合物由含有不饱和键的单体通过聚合得到。Preferably, the functionalized polymer is obtained by polymerization from a monomer containing an unsaturated bond.
优选地，所述含有不饱和键的单体为苯乙烯、甲基丙烯酸、2-甲基-2-丙烯酸-2，3-二羟基丙酯或甲基丙烯酸缩水甘油酯中的任意一种或至少两种的组合。Preferably, the unsaturated bond-containing monomer is any one of styrene, methacrylic acid, 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester or glycidyl methacrylate or A combination of at least two.
优选地，所述功能化聚合物的功能基团为羟基、氨基、羧基、巯基、醛基、环氧基或对甲苯磺酰基中的任意一种或至少两种的组合。Preferably, the functional group of the functionalized polymer is any one or a combination of at least two of a hydroxyl group, an amino group, a carboxyl group, a thiol group, an aldehyde group, an epoxy group or a p-toluenesulfonyl group.
本公开提供的磁性微球采用无孔聚合物微球而非多孔聚合物微球作为磁性微球的模板(内核)，避免了微球中的孔道在后续生物检测实验中可能造成对蛋白等生物大分子的非特异性吸附并导致背景噪声偏高。The magnetic microspheres provided by the present disclosure use non-porous polymer microspheres instead of porous polymer microspheres as a template (core) of magnetic microspheres, which avoids the pores in the microspheres may cause proteins and other organisms in subsequent bioassay experiments. Non-specific adsorption of macromolecules and high background noise.
第二方面，本公开提供了如上所述的磁性微球的制备方法，所述制备方法 包括如下步骤：In a second aspect, the present disclosure provides a method of preparing a magnetic microsphere as described above, the method comprising the steps of:
(1)在聚合物内核的表面进行聚合反应，得到具有刷状聚合物中间层的微球；(1) conducting a polymerization reaction on the surface of the polymer core to obtain a microsphere having a brush-like polymer intermediate layer;
(2)对步骤(1)得到的微球的刷状聚合物进行化学修饰，引入功能基团，得到带有功能基团的微球；(2) chemically modifying the brush-like polymer of the microsphere obtained in the step (1), introducing a functional group to obtain a microsphere with a functional group;
(3)在步骤(2)得到的带有功能基团的微球表面沉积磁性材料，得到带有磁性材料的微球；(3) depositing a magnetic material on the surface of the functional group-containing microsphere obtained in the step (2) to obtain a microsphere with a magnetic material;
(4)将步骤(3)得到的带有磁性材料的微球的表面进行聚合形成功能化聚合物外层，得到所述磁性微球。(4) The surface of the magnetic material-containing microsphere obtained in the step (3) is polymerized to form an outer layer of the functionalized polymer to obtain the magnetic microsphere.
在本公开中，通过对刷状聚合物进行功能化，可以引入高密度的功能基团，并以此为模板沉积磁性材料，制备磁性材料含量高的磁性微球。磁性材料可以通过共沉淀法制备：在表面带有功能基团的刷状聚合物模板的微球存在下，向微球分散液中加入可溶性金属盐，金属阳离子结合在刷状聚合物的功能基团上；然后，向分散液中加入碱液调节pH，金属阳离子从溶液中沉淀出来生成纳米尺寸的金属氧化物小颗粒。In the present disclosure, by functionalizing a brush-like polymer, a high-density functional group can be introduced, and a magnetic material can be deposited as a template to prepare a magnetic microsphere having a high magnetic material content. The magnetic material can be prepared by a coprecipitation method: a soluble metal salt is added to the microsphere dispersion in the presence of a microsphere of a brush-like polymer template having a functional group on the surface, and the metal cation is bonded to the functional group of the brush polymer. Then, an alkali solution is added to the dispersion to adjust the pH, and the metal cation is precipitated from the solution to form small-sized metal oxide small particles.
本公开提供的制备方法简单易行，利用本公开提供的制备方法制备得到的磁性微球尺寸均一且不具有孔道结构。尺寸均一，不具备孔道结构的磁性微球是开发稳定性好、背景噪声低、灵敏度高的诊断试剂盒的基础。The preparation method provided by the present disclosure is simple and easy, and the magnetic microspheres prepared by the preparation method provided by the present disclosure are uniform in size and have no pore structure. Magnetic microspheres with uniform dimensions and no pore structure are the basis for the development of diagnostic kits with good stability, low background noise and high sensitivity.
在本公开中，所述制备方法还包括通过聚合反应制备所述聚合物内核。In the present disclosure, the preparation method further includes preparing the polymer core by a polymerization reaction.
优选地，所述聚合反应为乳液聚合、微乳液聚合、细乳液聚合、无皂乳液聚合、分散聚合、悬浮聚合或种子聚合中的任意一种或至少两种的组合。Preferably, the polymerization reaction is any one or a combination of at least two of emulsion polymerization, microemulsion polymerization, miniemulsion polymerization, soap-free emulsion polymerization, dispersion polymerization, suspension polymerization or seed polymerization.
本公开的技术方案先合成聚合物内核微球再引入磁性材料，避免了在合成聚合物微球同时加入磁性材料导致微球尺寸不均一的问题，可以制得尺寸高度 均一的磁性微球，在后续的生物检测应用中具有更好的批次稳定性。The technical solution of the present disclosure first synthesizes the polymer core microspheres and then introduces the magnetic material, thereby avoiding the problem that the microspheres are not uniform in size by adding the magnetic materials while synthesizing the polymer microspheres, and the magnetic microspheres having a uniform height can be obtained. Better batch stability in subsequent bioassay applications.
本公开提供的磁性微球的刷状聚合物中间层提供大量的功能基团作为金属氧化物的结合位点，能有效提高磁性材料的沉积量，弥补了以无孔聚合物微球作为模板的磁性微球中磁性材料含量通常少于以多孔聚合物微球作为模板的磁性微球中磁性材料含量的缺点。The brush-like polymer intermediate layer of the magnetic microsphere provided by the present disclosure provides a large number of functional groups as a binding site of the metal oxide, can effectively increase the deposition amount of the magnetic material, and compensates for the non-porous polymer microsphere as a template. The magnetic material content of the magnetic microspheres is generally less than that of the magnetic material content of the magnetic microspheres using the porous polymer microspheres as a template.
在本公开中，作为优选技术方案，聚合物内核可以是聚苯乙烯微球，制备方法如下：In the present disclosure, as a preferred technical solution, the polymer core may be a polystyrene microsphere, and the preparation method is as follows:
I、将苯乙烯和交联剂加入溶剂中，搅拌溶解；I, adding styrene and a crosslinking agent to the solvent, stirring and dissolving;
II、将引发剂加入步骤I得到的溶剂中，搅拌溶解，加入乳化剂，混合；II, the initiator is added to the solvent obtained in the step I, stirred and dissolved, added with an emulsifier, and mixed;
III、将聚苯乙烯种子加入步骤II得到的乳液中，反应，得到聚苯乙烯微球。III. The polystyrene seeds are added to the emulsion obtained in the step II, and reacted to obtain polystyrene microspheres.
优选地，步骤I所述交联剂为二乙烯苯。Preferably, the crosslinking agent in step I is divinylbenzene.
优选地，步骤I所述溶剂为环己醇。Preferably, the solvent of step I is cyclohexanol.
优选地，步骤I所述苯乙烯、交联剂和溶剂的质量比为(25-35)∶(50-60)∶(100-120)，例如25∶50∶100、30∶55∶110、35∶60∶120、30∶60∶100等。Preferably, the mass ratio of the styrene, the crosslinking agent and the solvent in the step I is (25-35):(50-60):(100-120), for example 25:50:100, 30:55:110, 35:60:120, 30:60:100, and the like.
优选地，步骤II所述引发剂为偶氮二异丁腈，所述引发剂的用量为苯乙烯的3-5wt％，例如3wt％、4wt％、5wt％等。Preferably, the initiator in step II is azobisisobutyronitrile, and the initiator is used in an amount of 3-5 wt% of styrene, such as 3 wt%, 4 wt%, 5 wt%, and the like.
优选地，步骤II所述乳化剂为十八烷基磺酸钠。Preferably, the emulsifier of step II is sodium octadecyl sulfonate.
优选地，步骤III所述聚苯乙烯种子为粒径为0.005-1000μm的聚苯乙烯微球，所述0.005-1000μm可以是0.005μm、0.01μm、0.1μm、1μm、10μm、100μm、1000μm等。Preferably, the polystyrene seed in the step III is a polystyrene microsphere having a particle diameter of 0.005 to 1000 μm, and the 0.005 to 1000 μm may be 0.005 μm, 0.01 μm, 0.1 μm, 1 μm, 10 μm, 100 μm, 1000 μm or the like.
优选地，步骤III所述反应为在室温下反应24h后升温到80-100℃反应，所述80-100℃可以是80℃、85℃、90℃、95℃、100℃等。Preferably, the reaction in the step III is a reaction at room temperature for 24 hours and then the temperature is raised to 80-100 ° C. The 80-100 ° C may be 80 ° C, 85 ° C, 90 ° C, 95 ° C, 100 ° C, and the like.
优选地，步骤III得到的聚苯乙烯微球的粒径为0.01μm-1000μm，例如0.01 μm、0.1μm、1μm、10μm、100μm、1000μm等。Preferably, the polystyrene microspheres obtained in the step III have a particle diameter of 0.01 μm to 1000 μm, for example, 0.01 μm, 0.1 μm, 1 μm, 10 μm, 100 μm, 1000 μm, or the like.
在本公开中，作为优选技术方案，磁性微球的制备方法如下：In the present disclosure, as a preferred technical solution, the preparation method of the magnetic microspheres is as follows:
(1)将刷状聚合物单体加入溶剂中溶解得到溶液，加入催化剂并搅拌，得到反应溶液A；(1) adding a brush-like polymer monomer to a solvent to obtain a solution, adding a catalyst and stirring to obtain a reaction solution A;
(2)将聚合物内核加入溶剂中，超声，得到反应溶液B；(2) adding the polymer core to the solvent, ultrasonication, to obtain a reaction solution B;
(3)将反应溶液A和反应溶液B混合，反应，得到带有刷状聚合物中间层的微球；(3) mixing the reaction solution A and the reaction solution B, and reacting to obtain microspheres having a brush-like polymer intermediate layer;
(4)将步骤(3)得到的微球分散在溶剂中，加入功能单体，反应，得到带有功能基团的微球；(4) dispersing the microspheres obtained in the step (3) in a solvent, adding a functional monomer, and reacting to obtain microspheres having functional groups;
(5)将步骤(4)得到的带有功能基团的微球分散在溶剂中，表面沉积磁性材料，得到带有磁性材料的微球；(5) dispersing the functional group-containing microspheres obtained in the step (4) in a solvent, and depositing a magnetic material on the surface to obtain microspheres with a magnetic material;
(6)将步骤(5)得到的微球分散在溶剂中，加入聚合物单体和引发剂，进行聚合反应，形成功能化聚合物外层，得到磁性微球。(6) The microspheres obtained in the step (5) are dispersed in a solvent, a polymer monomer and an initiator are added, and polymerization is carried out to form an outer layer of the functionalized polymer to obtain magnetic microspheres.
优选地，步骤(1)所述刷状聚合物单体为甲基丙烯酸缩水甘油酯、甲基丙烯酸叔丁酯或2-甲基-2-丙烯酸-2，3-二羟基丙酯中的任意一种或至少两种的组合。Preferably, the brush-like polymer monomer in the step (1) is any one of glycidyl methacrylate, t-butyl methacrylate or 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester. One or a combination of at least two.
优选地，步骤(1)所述溶剂为异丙醇，所述刷状聚合物单体的添加量为1mol/L(溶液)。Preferably, the solvent in the step (1) is isopropyl alcohol, and the amount of the brush-like polymer monomer is 1 mol/L (solution).
优选地，步骤(1)所述催化剂包括氯化亚铜、氯化铜和三(2-二甲氨基乙基)胺，所述氯化亚铜、氯化铜和三(2-二甲氨基乙基)胺添加的质量比为(5-10)∶(1-2)∶(10-20)，例如5∶1∶10、8∶2∶15、10∶2∶20、7∶1∶20等。Preferably, the catalyst of the step (1) comprises cuprous chloride, cupric chloride and tris(2-dimethylaminoethyl)amine, the cuprous chloride, cupric chloride and tris(2-dimethylamino). The mass ratio of ethylamine addition is (5-10):(1-2):(10-20), for example 5:1:10, 8:2:15, 10:2:20, 7:1: 20 and so on.
优选地，步骤(1)为在氮气氛围下加入催化剂并搅拌。Preferably, step (1) is to add a catalyst under a nitrogen atmosphere and stir.
优选地，步骤(1)还包括在加入催化剂前，溶液通入氮气脱氧1-2小时， 例如1小时、1.5小时、2小时等。Preferably, step (1) further comprises deoxidizing the solution to nitrogen for 1-2 hours, for example 1 hour, 1.5 hours, 2 hours, etc., prior to the addition of the catalyst.
优选地，步骤(2)所述溶剂为异丙醇。Preferably, the solvent in the step (2) is isopropyl alcohol.
优选地，步骤(2)所述聚合物内核的添加量为50-60g/L(溶剂)，例如50g/L(溶剂)、52g/L(溶剂)、55g/L(溶剂)、57g/L(溶剂)、60g/L(溶剂)等。Preferably, the polymer core of the step (2) is added in an amount of 50-60 g/L (solvent), for example, 50 g/L (solvent), 52 g/L (solvent), 55 g/L (solvent), 57 g/L. (solvent), 60 g/L (solvent), and the like.
优选地，步骤(2)还包括在超声之后通入氮气脱氧1-2小时，例如1小时、1.5小时、2小时等。Preferably, step (2) further comprises deoxygenating with nitrogen for 1-2 hours after sonication, for example 1 hour, 1.5 hours, 2 hours, and the like.
优选地，步骤(3)所述反应的时间为24-30h，例如24h、26h、28h、30h等。Preferably, the reaction time in the step (3) is 24-30 h, such as 24 h, 26 h, 28 h, 30 h, and the like.
优选地，步骤(3)还包括通入空气停止反应。Preferably, step (3) further comprises introducing air to stop the reaction.
优选地，步骤(3)还包括反应结束后，对反应溶液进行过滤、洗涤和干燥。Preferably, the step (3) further comprises, after the reaction is completed, filtering, washing and drying the reaction solution.
优选地，步骤(4)所述微球的添加量为50-60g/L(溶剂)，例如50g/L(溶剂)、52g/L(溶剂)、55g/L(溶剂)、57g/L(溶剂)、60g/L(溶剂)等。Preferably, the microspheres in the step (4) are added in an amount of 50-60 g/L (solvent), for example, 50 g/L (solvent), 52 g/L (solvent), 55 g/L (solvent), 57 g/L ( Solvent), 60 g/L (solvent), and the like.
优选地，步骤(4)所述功能单体为乙二胺和/或盐酸。Preferably, the functional monomer in the step (4) is ethylenediamine and/or hydrochloric acid.
优选地，步骤(4)所述溶剂为无水乙醇和/或去离子水。Preferably, the solvent in the step (4) is anhydrous ethanol and/or deionized water.
优选地，步骤(4)所述反应为在室温下反应6-24h，例如6h、10h、15h、20h、24h等。Preferably, the reaction in the step (4) is a reaction at room temperature for 6 to 24 hours, for example, 6 hours, 10 hours, 15 hours, 20 hours, 24 hours, and the like.
优选地，步骤(4)还包括反应结束后对反应溶液进行过滤、洗涤和干燥。Preferably, the step (4) further comprises filtering, washing and drying the reaction solution after the reaction is completed.
优选地，步骤(5)所述溶剂为去离子水。Preferably, the solvent in the step (5) is deionized water.
优选地，步骤(5)所述带有功能基团的微球的添加量为15-25g/L(溶剂)，例如15g/L(溶剂)、17g/L(溶剂)、20g/L(溶剂)、22g/L(溶剂)、25g/L(溶剂)等。Preferably, the functional group-containing microspheres are added in an amount of 15-25 g/L (solvent), for example, 15 g/L (solvent), 17 g/L (solvent), 20 g/L (solvent). ), 22 g / L (solvent), 25 g / L (solvent) and the like.
优选地，步骤(5)所述沉积磁性材料的方法为在0-5℃下，将六水合氯化铁和/或四水合氯化亚铁加入微球溶液中，在氮气保护下搅拌，然后滴加氨水，滴加完成后，在室温下搅拌过夜，所述0-5℃可以是0℃、1℃、2℃、3℃、4℃、 5℃等。Preferably, the method of depositing the magnetic material in the step (5) is to add ferric chloride hexahydrate and/or ferrous chloride tetrahydrate to the microsphere solution at 0-5 ° C, stir under nitrogen, and then Ammonia water was added dropwise, and after completion of the dropwise addition, the mixture was stirred at room temperature overnight, and the 0-5 ° C may be 0 ° C, 1 ° C, 2 ° C, 3 ° C, 4 ° C, 5 ° C, and the like.
优选地，步骤(5)所述磁性材料的添加量为步骤(5)所述微球添加量的2-4倍，例如2倍、2.5倍、3倍、3.5倍、4倍等。Preferably, the amount of the magnetic material added in the step (5) is 2-4 times, that is, 2 times, 2.5 times, 3 times, 3.5 times, 4 times, etc., of the microspheres added in the step (5).
优选地，步骤(5)还包括表面沉积磁性材料结束后移去溶剂、洗涤，洗涤后重新分散在去离子水中。Preferably, step (5) further comprises removing the solvent after the surface deposition of the magnetic material, washing, washing and redispersing in deionized water.
优选地，步骤(6)所述溶剂为去离子水和Brij-35(30％)水溶液的混合液。Preferably, the solvent in the step (6) is a mixture of deionized water and a Brij-35 (30%) aqueous solution.
优选地，步骤(6)所述聚合物单体为苯乙烯、甲基丙烯酸、甲基丙烯酸缩水甘油酯或2-甲基-2-丙烯酸-2，3-二羟基丙酯中的任意一种或至少两种的组合。Preferably, the polymer monomer in the step (6) is any one of styrene, methacrylic acid, glycidyl methacrylate or 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester. Or a combination of at least two.
优选地，步骤(6)所述引发剂为过氧化(2-乙基己酸)叔丁酯。Preferably, the initiator in the step (6) is tert-butyl (2-ethylhexanoate) peroxide.
优选地，步骤(6)所述聚合反应在氮气氛围下进行。Preferably, the polymerization in the step (6) is carried out under a nitrogen atmosphere.
优选地，步骤(6)所述聚合反应的反应温度为80-100℃，例如80℃、85℃、90℃、95℃、100℃等。Preferably, the reaction temperature of the polymerization reaction in the step (6) is 80 to 100 ° C, for example, 80 ° C, 85 ° C, 90 ° C, 95 ° C, 100 ° C, and the like.
优选地，步骤(6)所述聚合反应的反应时间为8-10小时，例如8小时、8.5小时、9小时、9.5小时、10小时等。Preferably, the reaction time of the polymerization reaction in the step (6) is 8 to 10 hours, for example, 8 hours, 8.5 hours, 9 hours, 9.5 hours, 10 hours, and the like.
优选地，步骤(6)还包括聚合反应结束后对反应溶液进行洗涤，洗涤后重新分散在去离子水中。Preferably, step (6) further comprises washing the reaction solution after the end of the polymerization reaction, washing and redispersing in deionized water.
第三方面，本公开提供了如上所述的磁性微球在制备诊断试剂盒中的应用。In a third aspect, the present disclosure provides the use of a magnetic microsphere as described above in the preparation of a diagnostic kit.
尺寸均一的磁性微球在后续的生物检测应用中具有更好的批次稳定性，本公开提供的磁性微球尺寸均一，磁性材料含量高且不具备孔道结构，是开发稳定性好、背景噪声低、灵敏度高的诊断试剂盒的基础。Uniform magnetic microspheres have better batch stability in subsequent bio-detection applications. The magnetic microspheres provided by the present disclosure have uniform size, high magnetic material content and no pore structure, and are stable in development and background noise. The basis for a low-sensitivity diagnostic kit.
相对于现有技术，本公开具有以下有益效果：Compared with the prior art, the present disclosure has the following beneficial effects:
本公开提供的磁性微球为“三明治”型夹心结构的磁性微球，本公开提供的磁性微球中间层的刷状聚合物带有功能基团，功能基团作为金属氧化物的结 合位点，能有效提高磁性材料的沉积量，解决了以往磁性微球中磁性材料含量小的问题。本公开提供的制备方法简单易行。The magnetic microsphere provided by the present disclosure is a magnetic microsphere of a "sandwich" type sandwich structure, and the brush-like polymer of the intermediate layer of the magnetic microsphere provided by the present disclosure has a functional group, and the functional group serves as a binding site of the metal oxide. It can effectively increase the deposition amount of magnetic materials and solve the problem of small magnetic material content in magnetic microspheres in the past. The preparation method provided by the present disclosure is simple and easy.
具体实施方式Detailed ways
下面通过具体实施方式来进一步说明本公开的技术方案。本领域技术人员应该明了，所述实施例仅仅是帮助理解本发明，不应视为对本发明的具体限制。The technical solutions of the present disclosure will be further described below by way of specific embodiments. It should be understood by those skilled in the art that the present invention is not to be construed as limited.
在实施例1-2中所使用的聚合物内核为5μm的聚苯乙烯微球，苏州纳微科技有限公司生产，型号为UniPS-NP5。The polymer core used in Example 1-2 was a 5 μm polystyrene microsphere, manufactured by Suzhou Nawei Technology Co., Ltd., model No. UniPS-NP5.
实施例1Example 1
本实施例提供的磁性微球包括直径为5μm的聚苯乙烯微球内核、带有磁性氧化铁的刷状甲基丙烯酸缩水甘油酯的中间层和功能化聚合物外层，刷状甲基丙烯酸缩水甘油酯带有氨基。The magnetic microspheres provided in this embodiment include a polystyrene microsphere core having a diameter of 5 μm, an intermediate layer of brush-like glycidyl methacrylate with magnetic iron oxide, and an outer layer of a functionalized polymer, brush-like methacrylic acid. Glycidyl esters carry an amino group.
制备方法如下：The preparation method is as follows:
(1)聚合物内核表面活化，引入反应基团氯甲基(1) Surface activation of the polymer core, introduction of reactive group chloromethyl
将粒径为5μm的聚苯乙烯微球10.0g与100mL无水三氯甲烷加入到250mL三口烧瓶中，用机械搅拌将微球分散在溶剂中，分散的微球在0℃搅拌1小时，加入1.7mL无水四氯化锡，搅拌5分钟后逐滴加入8.0mL氯甲醚，滴加完毕后维持在0℃搅拌30分钟，然后升温至30℃继续搅拌3小时。反应结束后用砂芯漏斗过滤除去溶液，然后依次用200mL以下溶剂：去离子水、5％盐酸、去离子水、四氢呋喃、乙醇、丙酮，对微球进行清洗。清洗后的微球于60℃干燥12个小时，得到氯甲基化的微球。10.0 g of polystyrene microspheres having a particle diameter of 5 μm and 100 mL of anhydrous chloroform were placed in a 250 mL three-necked flask, and the microspheres were dispersed in a solvent by mechanical stirring, and the dispersed microspheres were stirred at 0 ° C for 1 hour, and added. 1.7 mL of anhydrous tin tetrachloride was added, and after stirring for 5 minutes, 8.0 mL of chloromethyl ether was added dropwise, and after the completion of the dropwise addition, the mixture was stirred at 0 ° C for 30 minutes, and then the temperature was raised to 30 ° C and stirring was continued for 3 hours. After the reaction was completed, the solution was removed by filtration through a sand core funnel, and then the microspheres were washed with 200 mL of the following solvent: deionized water, 5% hydrochloric acid, deionized water, tetrahydrofuran, ethanol, acetone. The washed microspheres were dried at 60 ° C for 12 hours to obtain chloromethylated microspheres.
(2)制备具有刷状聚甲基丙烯酸缩水甘油酯中间层的微球(2) Preparation of microspheres having a brush-like intermediate layer of glycidyl methacrylate
将单体甲基丙烯酸甘油酯溶解于异丙醇中，制备浓度为1mol/L的溶液。将100mL上述单体的异丙醇溶液加入250mL三口烧瓶中，在25℃下通入氮气脱 氧1小时，然后在氮气保护下依次加入115mg氯化亚铜、15.7mg氯化铜和300mg三(2-二甲氨基乙基)胺，机械搅拌15分钟，得到反应溶液A1。The monomer glyceryl methacrylate was dissolved in isopropyl alcohol to prepare a solution having a concentration of 1 mol/L. 100 mL of the above-mentioned monomeric isopropanol solution was placed in a 250 mL three-necked flask, deoxygenated with nitrogen at 25 ° C for 1 hour, and then 115 mg of cuprous chloride, 15.7 mg of copper chloride and 300 mg of tris(2) were sequentially added under nitrogen protection. -Dimethylaminoethyl)amine, mechanically stirred for 15 minutes to obtain a reaction solution A1.
另取一干燥的250mL三口烧瓶，加入1.0g步骤(1)得到的微球与20mL异丙醇，超声分散后在25℃下通入氮气脱氧1小时，得到反应溶液B1。Another dry 250 mL three-necked flask was taken, and 1.0 g of the microspheres obtained in the step (1) and 20 mL of isopropyl alcohol were added, and ultrasonically dispersed, and deoxidized by nitrogen gas at 25 ° C for 1 hour to obtain a reaction solution B1.
在氮气保护下，将反应溶液A1加入反应溶液B1中，引发聚合。在25℃下机械搅拌24小时后，向溶液中通入空气，终止反应。用砂芯漏斗过滤除去溶液，然后依次用50mL以下溶剂：甲醇、50mM EDTA水溶液(pH 8.0)、去离子水(两次)、丙酮，对微球进行清洗。清洗后的微球于60℃干燥12个小时，得到表面接枝刷状聚甲基丙烯酸缩水甘油酯的聚合物微球。The reaction solution A1 was added to the reaction solution B1 under a nitrogen atmosphere to initiate polymerization. After mechanical stirring at 25 ° C for 24 hours, air was bubbled through the solution to terminate the reaction. The solution was removed by filtration using a sand core funnel, and then the microspheres were washed with 50 mL of the following solvent: methanol, 50 mM aqueous EDTA (pH 8.0), deionized water (twice), and acetone. The washed microspheres were dried at 60 ° C for 12 hours to obtain polymer microspheres of surface-grafted brush-like polyglycidyl methacrylate.
(3)刷状聚甲基丙烯酸缩水甘油酯引入氨基(3) Brush-like polyglycidyl methacrylate is introduced into the amino group
将步骤(2)得到的微球1.0g超声分散在20mL无水乙醇中，加入5mL无水乙二胺，在室温下旋转反应6小时。用砂芯漏斗过滤除去溶液，用无水乙醇洗涤两次，每次50mL，再用去离子水洗涤两次，每次50mL，最后用50mL丙酮洗涤一次。清洗后的微球于60℃干燥12个小时，得到表面含氨基的微球。1.0 g of the microspheres obtained in the step (2) was ultrasonically dispersed in 20 mL of absolute ethanol, and 5 mL of anhydrous ethylenediamine was added thereto, and the reaction was rotated at room temperature for 6 hours. The solution was removed by filtration through a sand funnel, washed twice with absolute ethanol, 50 mL each time, and then washed twice with deionized water, 50 mL each time, and finally washed once with 50 mL of acetone. The washed microspheres were dried at 60 ° C for 12 hours to obtain microspheres having amino groups on the surface.
(4)沉积磁性氧化铁(4) Depositing magnetic iron oxide
将步骤(3)得到的微球1.0g加入250mL三口烧瓶中，向烧瓶加入50mL去离子水，超声分散微球。将烧瓶浸入冰水浴中，待溶液温度降至5℃以下，在氮气保护加机械搅拌下，向烧瓶中加入2.0g六水合氯化铁(III)和0.93g四水合氯化亚铁。在氮气保护下继续搅拌1小时，然后用滴液漏斗向溶液中缓慢加入8mL氨水。加入完成后，移去冰水浴，在室温下继续搅拌过夜。反应完成后，将沉积磁性氧化铁的微球用磁铁分离，移去上清液，然后用去离子水洗涤三次，每次50mL，最后分散在25mL去离子水中，得到带有磁性氧化铁的微球。微球分散液保存于2-8℃。1.0 g of the microspheres obtained in the step (3) was placed in a 250 mL three-necked flask, and 50 mL of deionized water was added to the flask to ultrasonically disperse the microspheres. The flask was immersed in an ice water bath, and the temperature of the solution was lowered to below 5 ° C. Under a nitrogen atmosphere plus mechanical stirring, 2.0 g of iron (III) chloride hexahydrate and 0.93 g of ferrous chloride tetrahydrate were added to the flask. Stirring was continued for 1 hour under nitrogen, and then 8 mL of aqueous ammonia was slowly added to the solution using a dropping funnel. After the addition was completed, the ice water bath was removed and stirring was continued at room temperature overnight. After the reaction is completed, the magnetic iron oxide-deposited microspheres are separated by a magnet, the supernatant is removed, and then washed three times with 50 mL of deionized water, and finally dispersed in 25 mL of deionized water to obtain micro-magnetic iron oxide. ball. The microsphere dispersion was stored at 2-8 °C.
(5)制备功能化聚合物外层，得到磁性微球(5) preparing a functionalized polymer outer layer to obtain magnetic microspheres
将步骤(4)得到的微球1.0g加入100mL三口烧瓶中，加入30mL去离子水和0.5mL Brij-35 30％水溶液，超声分散微球。向上述分散液中加入0.2g苯乙烯，在25℃下通入氮气脱氧1小时，然后加入0.05g引发剂过氧化(2-乙基己酸)叔丁酯，在氮气保护下升温至80℃引发聚合。反应8小时后，将反应体系冷却至室温，得到表面包被一层聚合物的夹心结构磁性微球。将产物用乙醇洗涤三次，每次50mL，然后超声分散在30mL含有0.5％Brij-35的水溶液中，再向分散液中加入0.2g苯乙烯、0.01g甲基丙烯酸和0.02g甲基丙烯酸缩水甘油酯，在25℃下通入氮气脱氧1小时，然后加入0.02g引发剂过氧化(2-乙基己酸)叔丁酯，在氮气保护下升温至80℃引发聚合。反应8小时后，将反应体系冷却至室温，得到表面包被两层聚合物的夹心结构磁性微球。将产物用乙醇洗涤两次，每次50mL，再用50mL去离子水洗涤一次，最后分散在25mL去离子水中，保存于2-8℃。1.0 g of the microspheres obtained in the step (4) was placed in a 100 mL three-necked flask, and 30 mL of deionized water and 0.5 mL of Brij-35 30% aqueous solution were added to ultrasonically disperse the microspheres. To the above dispersion was added 0.2 g of styrene, deoxygenated with nitrogen at 25 ° C for 1 hour, then 0.05 g of initiator (2-ethylhexanoic acid) tert-butyl ester was added, and the temperature was raised to 80 ° C under nitrogen atmosphere. Initiate polymerization. After reacting for 8 hours, the reaction system was cooled to room temperature to obtain a sandwich-structure magnetic microsphere having a surface coated with a polymer. The product was washed three times with ethanol, 50 mL each time, then ultrasonically dispersed in 30 mL of an aqueous solution containing 0.5% Brij-35, and 0.2 g of styrene, 0.01 g of methacrylic acid and 0.02 g of glycidol methacrylate were added to the dispersion. The ester was deoxygenated with nitrogen at 25 ° C for 1 hour, then 0.02 g of initiator (2-ethylhexanoic acid) tert-butyl ester was added, and the temperature was raised to 80 ° C under nitrogen atmosphere to initiate polymerization. After reacting for 8 hours, the reaction system was cooled to room temperature to obtain sandwich-structured magnetic microspheres having a surface coated with two layers of polymer. The product was washed twice with 50 mL each time, once with 50 mL of deionized water, and finally in 25 mL of deionized water and stored at 2-8 °C.
实施例2Example 2
本实施例提供的磁性微球包括直径为5μm的聚苯乙烯微球内核、带有磁性氧化铁的刷状甲基丙烯酸叔丁酯的中间层和功能化聚合物外层，刷状甲基丙烯酸叔丁酯带有羧基。The magnetic microsphere provided in this embodiment comprises a core of polystyrene microspheres having a diameter of 5 μm, an intermediate layer of brush-like tert-butyl methacrylate with magnetic iron oxide, and an outer layer of functionalized polymer, brush-like methacrylic acid. Tert-butyl ester has a carboxyl group.
制备方法如下：The preparation method is as follows:
(1)聚合物内核表面活化，引入反应基团氯甲基(1) Surface activation of the polymer core, introduction of reactive group chloromethyl
(2)制备具有刷状聚甲基丙烯酸叔丁酯中间层的微球(2) Preparation of microspheres having a brush-like intermediate layer of tert-butyl polymethacrylate
将单体甲基丙烯酸叔丁酯溶解于异丙醇中，制备浓度为1mol/L的溶液。将100mL上述单体的异丙醇溶液加入250mL三口烧瓶中，在25℃下通入氮气脱氧1小时，然后在氮气保护下依次加入115mg氯化亚铜、15.7mg氯化铜和300 mg三(2-二甲氨基乙基)胺，机械搅拌15分钟，得到反应溶液A2。The monomer tert-butyl methacrylate was dissolved in isopropyl alcohol to prepare a solution having a concentration of 1 mol/L. 100 mL of the above-mentioned monomeric isopropanol solution was placed in a 250 mL three-necked flask, deoxygenated with nitrogen at 25 ° C for 1 hour, and then 115 mg of cuprous chloride, 15.7 mg of copper chloride and 300 mg of tri 2-Dimethylaminoethyl)amine was mechanically stirred for 15 minutes to obtain a reaction solution A2.
另取一干燥的250mL三口烧瓶，加入1.0g步骤(1)得到的氯甲基化聚合物内核微球与20mL异丙醇，超声分散后在25℃下通入氮气脱氧1小时，得到反应溶液B2。Another dry 250 mL three-necked flask was taken, 1.0 g of the chloromethylated polymer core microspheres obtained in the step (1) and 20 mL of isopropyl alcohol were added, and ultrasonically dispersed, and deoxidized by nitrogen gas at 25 ° C for 1 hour to obtain a reaction solution. B2.
在氮气保护下，将反应溶液A2加入反应溶液B2中，引发聚合。在25℃下机械搅拌24小时后，向溶液中通入空气，终止反应。用砂芯漏斗过滤除去溶液，然后依次用50mL以下溶剂：甲醇、50mM EDTA水溶液(pH 8.0)、去离子水(两次)、丙酮，对微球进行清洗。清洗后的微球于60℃干燥12个小时，得到表面接枝刷状聚甲基丙烯酸叔丁酯的聚合物微球。The reaction solution A2 was added to the reaction solution B2 under a nitrogen atmosphere to initiate polymerization. After mechanical stirring at 25 ° C for 24 hours, air was bubbled through the solution to terminate the reaction. The solution was removed by filtration using a sand core funnel, and then the microspheres were washed with 50 mL of the following solvent: methanol, 50 mM aqueous EDTA (pH 8.0), deionized water (twice), and acetone. The washed microspheres were dried at 60 ° C for 12 hours to obtain polymer microspheres of surface-grafted brush-like polybutyl methacrylate.
(3)刷状聚甲基丙烯酸叔丁酯引入羧基(3) Brush-like polybutyl methacrylate is introduced into the carboxyl group
将步骤(2)得到的微球1.0g超声分散在20mL去离子水中，用6mol/L盐酸调节pH至1。在室温下旋转反应过夜。用砂芯漏斗过滤除去溶液，用0.1mol/L氢氧化钠溶液洗涤两次，每次50mL，再用去离子水洗涤，每次50mL，直至滤液pH为中性，最后用50mL丙酮洗涤一次。清洗后的微球于60℃干燥12个小时，得到含羧基的微球。1.0 g of the microspheres obtained in the step (2) was ultrasonically dispersed in 20 mL of deionized water, and the pH was adjusted to 1 with 6 mol/L hydrochloric acid. The reaction was rotated overnight at room temperature. The solution was removed by filtration through a sand core funnel, washed twice with 0.1 mol/L sodium hydroxide solution, 50 mL each time, and then washed with deionized water, 50 mL each time until the filtrate pH was neutral, and finally washed once with 50 mL of acetone. The washed microspheres were dried at 60 ° C for 12 hours to obtain carboxyl group-containing microspheres.
(4)沉积磁性氧化铁(4) Depositing magnetic iron oxide
将步骤(3)得到的微球1.0g加入250mL三口烧瓶中，向烧瓶加入50mL去离子水，超声分散微球。将烧瓶浸入冰水浴中，待溶液温度降至5℃以下，在氮气保护加机械搅拌下，向烧瓶中加入2.0g六水合氯化铁(III)和0.93g四水合氯化亚铁。在氮气保护下继续搅拌1小时，然后用滴液漏斗向溶液中缓慢加入8mL氨水。加入完成后，移去冰水浴，在室温下继续搅拌过夜。反应完成后，将沉积磁性氧化铁的微球用磁铁分离，移去上清液，然后用去离子水洗涤三次，每次50mL，最后分散在25mL去离子水中，得到带有磁性氧化铁的微球。微 球分散液保存于2-8℃。1.0 g of the microspheres obtained in the step (3) was placed in a 250 mL three-necked flask, and 50 mL of deionized water was added to the flask to ultrasonically disperse the microspheres. The flask was immersed in an ice water bath, and the temperature of the solution was lowered to below 5 ° C. Under a nitrogen atmosphere plus mechanical stirring, 2.0 g of iron (III) chloride hexahydrate and 0.93 g of ferrous chloride tetrahydrate were added to the flask. Stirring was continued for 1 hour under nitrogen, and then 8 mL of aqueous ammonia was slowly added to the solution using a dropping funnel. After the addition was completed, the ice water bath was removed and stirring was continued at room temperature overnight. After the reaction is completed, the magnetic iron oxide-deposited microspheres are separated by a magnet, the supernatant is removed, and then washed three times with 50 mL of deionized water, and finally dispersed in 25 mL of deionized water to obtain micro-magnetic iron oxide. ball. The microsphere dispersion was stored at 2-8 °C.
(5)制备功能化聚合物外层，得到磁性微球(5) preparing a functionalized polymer outer layer to obtain magnetic microspheres
将步骤(4)得到的微球1.0g加入100mL三口烧瓶中，加入30mL去离子水和0.5mL Brij-35 30％水溶液，超声分散微球。向上述分散液中加入0.2g苯乙烯和0.01g甲基丙烯酸，在25℃下通入氮气脱氧1小时，然后加入0.02g引发剂过氧化(2-乙基己酸)叔丁酯，在氮气保护下升温至80℃引发聚合。反应8小时后，将反应体系冷却至室温，得到表面包被一层聚合物的磁性微球。将产物用乙醇洗涤两次，每次50mL，再用50mL去离子水洗涤一次，最后分散在25mL去离子水中，保存于2-8℃。1.0 g of the microspheres obtained in the step (4) was placed in a 100 mL three-necked flask, and 30 mL of deionized water and 0.5 mL of Brij-35 30% aqueous solution were added to ultrasonically disperse the microspheres. To the above dispersion, 0.2 g of styrene and 0.01 g of methacrylic acid were added, and deoxygenated with nitrogen at 25 ° C for 1 hour, and then 0.02 g of an initiator (2-ethylhexanoic acid) tert-butyl ester was added thereto, under nitrogen. The temperature was raised to 80 ° C under protection to initiate polymerization. After reacting for 8 hours, the reaction system was cooled to room temperature to obtain magnetic microspheres having a surface coated with a layer of a polymer. The product was washed twice with 50 mL each time, once with 50 mL of deionized water, and finally in 25 mL of deionized water and stored at 2-8 °C.
实施例3Example 3
本实施例提供的磁性微球包括直径为5μm的聚苯乙烯微球内核、带有磁性氧化铁的刷状2-甲基-2-丙烯酸-2，3-二羟基丙酯的中间层和功能化聚合物外层，刷状2-甲基-2-丙烯酸-2，3-二羟基丙酯带有羟基。The magnetic microspheres provided in this embodiment include a polystyrene microsphere core having a diameter of 5 μm, an intermediate layer and a function of brush-like 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester with magnetic iron oxide. The outer layer of the polymer is brushed with 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester with a hydroxyl group.
(1)制备直径为5μm的聚苯乙烯微球内核(1) Preparation of a polystyrene microsphere core having a diameter of 5 μm
取30g苯乙烯，60g二乙烯苯(80％)和100g环己醇加入到2000mL三口烧瓶中，向其中加入引发剂1.5g偶氮二异丁腈，机械搅拌使之完全溶解。在上面溶液中加入800g含0.04％十八烷基磺酸钠水溶液，超声乳化后加入25g，2.5μm聚苯乙烯种子，在室温溶胀24h后加热到80℃反应。冷却后分离出交联聚苯乙烯微球作为聚合物微球内核，粒径5μm，变异系数CV＝3.5％。30 g of styrene, 60 g of divinylbenzene (80%) and 100 g of cyclohexanol were placed in a 2000 mL three-necked flask, to which 1.5 g of azobisisobutyronitrile was added, and mechanically stirred to completely dissolve. 800 g of an aqueous solution containing 0.04% sodium octadecylsulfonate was added to the above solution, and after ultrasonication, 25 g of 2.5 μm polystyrene seeds were added, and the mixture was swollen at room temperature for 24 hours and then heated to 80 ° C for reaction. After cooling, the crosslinked polystyrene microspheres were separated as the core of the polymer microspheres, and the particle size was 5 μm, and the coefficient of variation CV was 3.5%.
(2)聚合物内核表面活化，引入反应基团氯甲基(2) Surface activation of the polymer core, introduction of reactive group chloromethyl
(3)制备具有聚2-甲基-2-丙烯酸-2，3-二羟基丙酯中间层的微球(3) Preparation of microspheres having an intermediate layer of poly-2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester
将2-甲基-2-丙烯酸-2，3-二羟基丙酯溶解于异丙醇中，制备浓度为1mol/L的溶液。将100mL上述单体的异丙醇溶液加入250mL三口烧瓶中，在25℃下 通入氮气脱氧1小时，然后在氮气保护下依次加入115mg氯化亚铜、15.7mg氯化铜和300mg三(2-二甲氨基乙基)胺，机械搅拌15分钟，得到反应溶液A3。A solution of 2 mol/L was prepared by dissolving 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester in isopropanol. 100 mL of the above-mentioned monomeric isopropanol solution was placed in a 250 mL three-necked flask, deoxygenated with nitrogen at 25 ° C for 1 hour, and then 115 mg of cuprous chloride, 15.7 mg of copper chloride and 300 mg of tris(2) were sequentially added under nitrogen protection. -Dimethylaminoethyl)amine, mechanically stirred for 15 minutes to obtain a reaction solution A3.
另取一干燥的250mL三口烧瓶，加入1.0g步骤(2)得到的氯甲基化聚合物内核微球与20mL异丙醇，超声分散后在25℃下通入氮气脱氧1小时，得到反应溶液B3。在氮气保护下，将反应溶液A3加入反应溶液B3中，引发聚合。在25℃下机械搅拌24小时后，向溶液中通入空气，终止反应。用砂芯漏斗过滤除去溶液，然后依次用50mL以下溶剂：甲醇、50mM EDTA水溶液(pH 8.0)、去离子水(两次)、丙酮，对微球进行清洗。清洗后的微球于60℃干燥12个小时，得到表面接枝刷状聚(2-甲基-2-丙烯酸-2，3-二羟基丙酯)的聚合物微球。Another dry 250 mL three-necked flask was taken, 1.0 g of the chloromethylated polymer core microspheres obtained in the step (2) and 20 mL of isopropyl alcohol were added, and ultrasonically dispersed, and deoxidized by nitrogen gas at 25 ° C for 1 hour to obtain a reaction solution. B3. The reaction solution A3 was added to the reaction solution B3 under a nitrogen atmosphere to initiate polymerization. After mechanical stirring at 25 ° C for 24 hours, air was bubbled through the solution to terminate the reaction. The solution was removed by filtration using a sand core funnel, and then the microspheres were washed with 50 mL of the following solvent: methanol, 50 mM aqueous EDTA (pH 8.0), deionized water (twice), and acetone. The washed microspheres were dried at 60 ° C for 12 hours to obtain polymer microspheres having a surface grafted brush-like poly(2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester).
(4)沉积磁性氧化铁(4) Depositing magnetic iron oxide
将步骤(3)得到的微球1.0g加入250mL三口烧瓶中，向烧瓶加入50mL去离子水，超声分散微球。将烧瓶浸入冰水浴中，待溶液温度降至5℃以下，在氮气保护加机械搅拌下，向烧瓶中加入2.0g六水合氯化铁(III)和0.93g四水合氯化亚铁。在氮气保护下继续搅拌1小时，然后用滴液漏斗向溶液中缓慢加入8mL氨水。加入完成后，移去冰水浴，在室温下继续搅拌过夜。反应完成后，将沉积磁性氧化铁的微球用磁铁分离，移去上清液，然后用去离子水洗涤三次，每次50mL，最后分散在25mL去离子水中，得到带有磁性氧化铁的微球。微球分散液保存于2-8℃。1.0 g of the microspheres obtained in the step (3) was placed in a 250 mL three-necked flask, and 50 mL of deionized water was added to the flask to ultrasonically disperse the microspheres. The flask was immersed in an ice water bath, and the temperature of the solution was lowered to below 5 ° C. Under a nitrogen atmosphere plus mechanical stirring, 2.0 g of iron (III) chloride hexahydrate and 0.93 g of ferrous chloride tetrahydrate were added to the flask. Stirring was continued for 1 hour under nitrogen, and then 8 mL of aqueous ammonia was slowly added to the solution using a dropping funnel. After the addition was completed, the ice water bath was removed and stirring was continued at room temperature overnight. After the reaction is completed, the magnetic iron oxide-deposited microspheres are separated by a magnet, the supernatant is removed, and then washed three times with 50 mL of deionized water, and finally dispersed in 25 mL of deionized water to obtain micro-magnetic iron oxide. ball. The microsphere dispersion was stored at 2-8 °C.
(5)制备功能化聚合物外层，得到磁性微球(5) preparing a functionalized polymer outer layer to obtain magnetic microspheres
将步骤(4)得到的微球1.0g加入100mL三口烧瓶中，加入30mL去离子水和0.5mL Brij-35 30％水溶液，超声分散微球。向上述分散液中加入0.2g苯乙烯、0.01g甲基丙烯酸和0.02g 2-甲基-2-丙烯酸-2，3-二羟基丙酯，在25℃下通入氮气脱氧1小时，然后加入0.02g引发剂过氧化(2-乙基己酸)叔丁酯，在 氮气保护下升温至80℃引发聚合。反应8小时后，将反应体系冷却至室温，得到表面包被一层聚合物的磁性微球。将产物用乙醇洗涤两次，每次50mL，再用50mL去离子水洗涤一次，最后分散在25mL去离子水中，保存于2-8℃。1.0 g of the microspheres obtained in the step (4) was placed in a 100 mL three-necked flask, and 30 mL of deionized water and 0.5 mL of Brij-35 30% aqueous solution were added to ultrasonically disperse the microspheres. 0.2 g of styrene, 0.01 g of methacrylic acid and 0.02 g of 2,3-dihydroxypropyl 2-methyl-2-acrylate were added to the above dispersion, and deoxygenated under nitrogen at 25 ° C for 1 hour, and then added. 0.02 g of initiator, tert-butyl peroxy (2-ethylhexanoate), was heated to 80 ° C under nitrogen to initiate polymerization. After reacting for 8 hours, the reaction system was cooled to room temperature to obtain magnetic microspheres having a surface coated with a layer of a polymer. The product was washed twice with 50 mL each time, once with 50 mL of deionized water, and finally in 25 mL of deionized water and stored at 2-8 °C.
申请人声明，本公开通过上述实施例来说明本发明的磁性微球及其制备方法和应用，但本发明并不局限于上述实施例，即不意味着本发明必须依赖上述实施例才能实施。所属技术领域的技术人员应该明了，对本公开的任何改进，对本公开产品各原料的等效替换及辅助成分的添加、具体方式的选择等，均落在本发明的保护范围和公开范围之内。The Applicant declares that the present disclosure describes the magnetic microspheres of the present invention and the preparation method and application thereof by the above embodiments, but the present invention is not limited to the above embodiments, and does not mean that the present invention must be implemented by relying on the above embodiments. It should be apparent to those skilled in the art that any modifications of the present disclosure, equivalent substitutions of the various materials of the present disclosure, and the addition of the auxiliary components, the selection of the specific manners, and the like, are all within the scope of the present invention.

Claims (11)

  1. 一种磁性微球，其包括聚合物内核、带有磁性材料的刷状聚合物中间层和功能化聚合物外层，所述刷状聚合物带有功能基团。A magnetic microsphere comprising a polymeric core, a brushed polymeric intermediate layer with a magnetic material, and a functionalized polymeric outer layer, the brushed polymer having functional groups.
  2. 根据权利要求1所述的磁性微球，所述聚合物内核的直径为0.01μm-1000μm。The magnetic microsphere according to claim 1, wherein the polymer core has a diameter of from 0.01 μm to 1000 μm.
  3. 根据权利要求1或2所述的磁性微球，所述聚合物内核由带有不饱和键的单体通过聚合得到。The magnetic microsphere according to claim 1 or 2, wherein the polymer core is obtained by polymerization of a monomer having an unsaturated bond.
  4. 根据权利要求3所述的磁性微球，其中，所述聚合物内核由含有不饱和键的芳香族单体通过聚合得到；The magnetic microsphere according to claim 3, wherein the polymer core is obtained by polymerization from an aromatic monomer having an unsaturated bond;
    优选地，所述聚合物内核为聚苯乙烯微球。Preferably, the polymer core is a polystyrene microsphere.
  5. 根据权利要求3所述的磁性微球，其中，所述聚合物内核由含有不饱和键的丙烯酸酯或甲基丙烯酸酯类单体通过聚合得到。The magnetic microsphere according to claim 3, wherein the polymer core is obtained by polymerization of an acrylate or methacrylate monomer containing an unsaturated bond.
  6. 根据权利要求1所述的磁性微球，其中，所述刷状聚合物由含有不饱和键的丙烯酸酯或甲基丙烯酸酯类单体通过聚合得到；The magnetic microsphere according to claim 1, wherein the brush polymer is obtained by polymerization of an acrylate or methacrylate monomer containing an unsaturated bond;
    优选地，所述刷状聚合物为聚甲基丙烯酸缩水甘油酯、聚甲基丙烯酸叔丁酯或聚2-甲基-2-丙烯酸-2，3-二羟基丙酯中的任意一种或至少两种的组合；Preferably, the brush polymer is any one of polyglycidyl methacrylate, poly-tert-butyl methacrylate or poly-2,2-dihydroxypropyl 2-methyl-2-acrylate. a combination of at least two;
    优选地，所述刷状聚合物带有的功能基团为羟基、氨基、羧基、巯基或环氧基中的任意一种或至少两种的组合。Preferably, the brush-like polymer carries a functional group of any one of a hydroxyl group, an amino group, a carboxyl group, a thiol group or an epoxy group or a combination of at least two.
  7. 根据权利要求1所述的磁性微球，其中，所述磁性材料为Fe 3O 4、γ-Fe 2O 3或MFe 2O 4中的任意一种或至少两种的组合，M为除Fe以外的金属； The magnetic microsphere according to claim 1, wherein the magnetic material is any one or a combination of at least two of Fe 3 O 4 , γ-Fe 2 O 3 or MFe 2 O 4 , and M is Fe. Metal other than
    优选地，所述MFe 2O 4中的M为Co、Mg、Ni、Mn或Zn中的任意一种或至少两种的组合。 Preferably, M in the MFe 2 O 4 is any one of Co, Mg, Ni, Mn or Zn or a combination of at least two.
  8. 根据权利要求1所述的磁性微球，其中，所述功能化聚合物外层包括至少一层功能化聚合物；The magnetic microsphere of claim 1 wherein said functionalized polymer outer layer comprises at least one layer of a functionalized polymer;
    优选地，所述功能化聚合物由含有不饱和键的单体通过聚合得到；Preferably, the functionalized polymer is obtained by polymerization from a monomer having an unsaturated bond;
    优选地，所述含有不饱和键的单体为苯乙烯、甲基丙烯酸、2-甲基-2-丙烯酸-2，3-二羟基丙酯或甲基丙烯酸缩水甘油酯中的任意一种或至少两种的组合；Preferably, the unsaturated bond-containing monomer is any one of styrene, methacrylic acid, 2-methyl-2-acrylic acid-2,3-dihydroxypropyl ester or glycidyl methacrylate or a combination of at least two;
    优选地，所述功能化聚合物的功能基团为羟基、氨基、羧基、巯基、醛基、环氧基或对甲苯磺酰基中的任意一种或至少两种的组合。Preferably, the functional group of the functionalized polymer is any one or a combination of at least two of a hydroxyl group, an amino group, a carboxyl group, a thiol group, an aldehyde group, an epoxy group or a p-toluenesulfonyl group.
  9. 根据权利要求1-8中的任一项所述的磁性微球的制备方法，其中，所述制备方法包括如下步骤：The method for producing magnetic microspheres according to any one of claims 1 to 8, wherein the preparation method comprises the following steps:
    (1)在聚合物内核的表面进行聚合反应，得到具有刷状聚合物中间层的微球；(1) conducting a polymerization reaction on the surface of the polymer core to obtain a microsphere having a brush-like polymer intermediate layer;
    (2)对步骤(1)得到的微球的刷状聚合物进行化学修饰，引入功能基团，得到带有功能基团的微球；(2) chemically modifying the brush-like polymer of the microsphere obtained in the step (1), introducing a functional group to obtain a microsphere with a functional group;
    (3)在步骤(2)得到的带有功能基团的微球表面沉积磁性材料，得到带有磁性材料的微球；(3) depositing a magnetic material on the surface of the functional group-containing microsphere obtained in the step (2) to obtain a microsphere with a magnetic material;
    (4)将步骤(3)得到的带有磁性材料的微球的表面进行聚合形成功能化聚合物外层，得到所述磁性微球。(4) The surface of the magnetic material-containing microsphere obtained in the step (3) is polymerized to form an outer layer of the functionalized polymer to obtain the magnetic microsphere.
  10. 根据权利要求9所述的制备方法，还包括通过聚合反应制备所述聚合物内核；The production method according to claim 9, further comprising preparing the polymer core by polymerization;
    优选地，所述聚合反应为乳液聚合、微乳液聚合、细乳液聚合、无皂乳液聚合、分散聚合、悬浮聚合或种子聚合中的任意一种或至少两种的组合。Preferably, the polymerization reaction is any one or a combination of at least two of emulsion polymerization, microemulsion polymerization, miniemulsion polymerization, soap-free emulsion polymerization, dispersion polymerization, suspension polymerization or seed polymerization.
  11. 根据权利要求1-8中的任一项所述的磁性微球在制备诊断试剂盒中的应用。Use of the magnetic microspheres according to any one of claims 1-8 in the preparation of a diagnostic kit.
PCT/CN2018/074903 2017-12-25 2018-02-01 Magnetic microsphere and preparation method and application thereof WO2019127836A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201711421866.X 2017-12-25
CN201711421866.XA CN108129614B (en) 2017-12-25 2017-12-25 Magnetic microsphere and preparation method and application thereof

Publications (1)

Publication Number Publication Date
WO2019127836A1 true WO2019127836A1 (en) 2019-07-04

Family

ID=62392493

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2018/074903 WO2019127836A1 (en) 2017-12-25 2018-02-01 Magnetic microsphere and preparation method and application thereof

Country Status (2)

Country Link
CN (1) CN108129614B (en)
WO (1) WO2019127836A1 (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110106170B (en) * 2019-06-14 2021-02-05 洛阳爱森生物科技有限公司 Preparation method of nano biological magnetic beads for whole blood DNA extraction
CN110935405B (en) * 2019-11-11 2022-04-12 湖北新纵科病毒疾病工程技术有限公司 Preparation method of magnetic polystyrene microspheres with surfaces rich in double bonds
CN111569844B (en) * 2020-04-29 2023-05-26 南京理工大学 Preactivated hydrophilic magnetic microsphere and preparation method thereof
CN112034164B (en) * 2020-08-28 2022-05-27 武汉生之源生物科技股份有限公司 Chemiluminescence immune magnetic ball and preparation method thereof
CN115228450B (en) * 2022-07-12 2023-10-17 南京大学 Preparation method of magnetic ester-based resin with uniform particle size
CN115193419B (en) * 2022-09-13 2022-11-22 苏州纳微生命科技有限公司 Magnetic microsphere, preparation method, application and diagnostic kit thereof
CN116813654B (en) * 2023-08-25 2023-12-01 苏州纳微生命科技有限公司 Fluorescent dye, magnetic fluorescent coding microsphere, and preparation method and application thereof

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1348731A1 (en) * 2002-03-25 2003-10-01 JSR Corporation Process for producing particles for diagnostic reagent
CN1943560A (en) * 2005-10-09 2007-04-11 北京师范大学 Sandwich structure magnetic composite micro ball with functional shell layer, its preparing method and use
WO2007093451A2 (en) * 2006-02-17 2007-08-23 Topass Gmbh Multimodal imaging using a three compartment polymer nanoparticle with cell specificity
US20090169866A1 (en) * 2007-12-31 2009-07-02 Agnes Ostafin Nanocomposite materials with dynamically adjusting refractive index and methods of making the same
CN101817960A (en) * 2010-03-17 2010-09-01 中国石油大学(华东) Method for preparing magnetic composite nanoparticles with core-shell structure
CN102990062A (en) * 2012-12-17 2013-03-27 苏州纳微生物科技有限公司 Method for preparing composite microsphere

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106540668B (en) * 2015-09-18 2019-04-23 中华人民共和国淮安出入境检验检疫局 Magnetic hydrophilic molecules trace composite material and preparation method

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1348731A1 (en) * 2002-03-25 2003-10-01 JSR Corporation Process for producing particles for diagnostic reagent
CN1943560A (en) * 2005-10-09 2007-04-11 北京师范大学 Sandwich structure magnetic composite micro ball with functional shell layer, its preparing method and use
WO2007093451A2 (en) * 2006-02-17 2007-08-23 Topass Gmbh Multimodal imaging using a three compartment polymer nanoparticle with cell specificity
US20090169866A1 (en) * 2007-12-31 2009-07-02 Agnes Ostafin Nanocomposite materials with dynamically adjusting refractive index and methods of making the same
CN101817960A (en) * 2010-03-17 2010-09-01 中国石油大学(华东) Method for preparing magnetic composite nanoparticles with core-shell structure
CN102990062A (en) * 2012-12-17 2013-03-27 苏州纳微生物科技有限公司 Method for preparing composite microsphere

Also Published As

Publication number Publication date
CN108129614A (en) 2018-06-08
CN108129614B (en) 2020-11-24

Similar Documents

Publication Publication Date Title
WO2019127836A1 (en) Magnetic microsphere and preparation method and application thereof
US4873102A (en) Magnetic particles
CN111375360B (en) Preparation method of magnetic microspheres with uniform particle size
CN110396148B (en) Magnetic polystyrene microsphere and preparation method thereof
CN111393574B (en) Magnetic microsphere with functional groups on surface and preparation method and application thereof
CN112666140B (en) Poly (undecylenic acid-divinylbenzene) -coated magnetic fluorescently encoded microspheres
CN116082669A (en) Polymer-based nano magnetic beads and preparation method thereof
CN114591726B (en) Method for preparing stable monodisperse crosslinked polystyrene magnetic microspheres
CN112175150A (en) Novel preparation method of functionalized porous magnetic microspheres
CN108192081B (en) Preparation method of epoxy resin coated magnetic beads
CN108314868A (en) A kind of lignin composite hydrogel and preparation method thereof
CN115304882B (en) Preparation method of polymer coated porous magnetic polymer microsphere with controllable topological structure on surface
CN114106254B (en) Method for preparing functionalized magnetic polymer microsphere by miniemulsion polymerization method using porous microsphere as template
CN106883360B (en) Functionalized stimulus-responsive polymer and preparation method thereof
CN115746389A (en) Amino-sulfonic acid group difunctional magnetic polymer microsphere and preparation method thereof
CN110739110B (en) Spherical nano magneton with stable solution dispersibility and preparation method thereof
CN110935405B (en) Preparation method of magnetic polystyrene microspheres with surfaces rich in double bonds
JPH08176461A (en) Fine monodisperse particle and its production
CN114100535A (en) Preparation method and application of monodisperse magnetic polydopamine microsphere
JP2023524994A (en) Monodisperse superparamagnetic particles and method for producing the same
CN114887559B (en) Preparation method of magnetic microsphere coated by poly (amino ester) with carboxyl group
CN115710373B (en) Preparation method of hydrogel magnetic microsphere
CN115975252B (en) Preparation method of magnetic polymer microsphere with double-layer shell structure
CN111116851A (en) Preparation method of polyquaternium magnetic microsphere with core-shell structure
CN110732296B (en) Preparation method of magnetic microspheres with fluffy shell layers

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18896627

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 18896627

Country of ref document: EP

Kind code of ref document: A1