JPH1048213A - Novel macromolecular carrier and production thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a magnetic particle excellent in characteristics, e.g. uniform size, and a convenient production method thereof without dissociating a magnetic body by embedding the magnetic body in the surface layer of macromolecular material utilizing the physical chemical properties of macromolecular material. SOLUTION: A magnetic body or a labeling substance is present in the gap of molecules composing a macromolecular material while being embedded in the surface layer thereof. The embedding part is not limited specially but it is normally set in the range of 25%, more preferably 15%, from the surface layer of macromolecular material. The macromolecular material is a synthetic macromolecular substance having a carboxyl group or an amino group and swelling in an alkaline or acid aqueous solvent. Any compound may be employed as a polymerization monomer having a carboxyl group or an amino group so long as a carboxyl group or an amino group is provided as a functional group which can be substituted by a lower alkyl group and a conjugated double bond is provided so that a polymer can be formed.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel polymer carrier and a method for producing the same, and more particularly, to a method of embedding a magnetic substance or a labeling substance inside a surface layer of a material having a carboxyl group or an amino group. And a method for producing the same.

[0002]

2. Description of the Related Art Magnetic particulate carriers (hereinafter sometimes abbreviated as "magnetic particles") are used for recording media such as magnetic tapes, and require washing, collecting and separating operations. Because it is extremely easy, it is used in various formulation and diagnostic techniques. These magnetic particles are mainly composed of (1) a method in which an emulsion is formed on the surface of a magnetic substance as a nucleus and polymerized (JP-A-56-164503), and (2) a method in which the surface of the particle is coated with a magnetic substance. And so on.

However, the magnetic particles synthesized by the method (1) have a shape in which the magnetic substance is coated with a polymer, and the magnetic particles serving as nuclei have different particle diameters.
The resulting magnetic particles have different sizes. For this reason, it is difficult to control the size of the magnetic particles uniformly, and it is particularly difficult to control the size of the magnetic material within the range of 0.1 to 1.0 μm. Also, the synthesis operation is very complicated. On the other hand, in the method (2), the magnetic material is coated on the surface of the carrier, but the magnetic material is adhered, so that the magnetic material is dissociated. Various techniques have been employed to prevent this dissociation, but the nature of the carrier surface is often different from the initial properties due to this.

As described above, the conventional method for producing magnetic particles has many problems, and there is a need for magnetic particles of higher quality and a method for producing the same.

[0005]

DISCLOSURE OF THE INVENTION The present invention solves the above-mentioned problems, that is, magnetic particles having excellent properties such as uniform size without dissociation of the magnetic material and a simple method for producing the same. It was made for the purpose of providing.

[0006]

Means for Solving the Problems The inventors of the present invention have conducted intensive studies to solve the above-mentioned problems, and as a result, the physico-chemical properties of the polymer material inside the surface layer of the polymer material having a carboxyl group or an amino group have been found. By embedding a magnetic material using its properties,
Magnetic particles can be easily produced, and the embedded polymer carrier having the magnetic substance thus produced is a magnetic particle having a uniform particle size and excellent properties such as no dissociation of the magnetic substance. I found it. The present invention has been completed based on this finding.

That is, the present invention provides an embedded polymer carrier in which a magnetic substance or a labeling substance is embedded inside the surface layer of a polymer material having a carboxyl group or an amino group. According to a preferred aspect of the present invention, the carrier, wherein the polymer material is a material made of a synthetic polymer material;
Synthetic polymer substance is polyacrylic acid or acrylic acid-
The above carrier, which is a styrene copolymer; the above carrier, wherein the polymer material is a spherical body;
the above-mentioned carrier in the range of μm; the above-mentioned carrier in which the magnetic substance or the labeling substance is embedded within the range of 25% from the surface layer of the polymer material; the polymer carrier is a bifunctional monomer and / or polyfunctional The above carrier, wherein the bifunctional monomer is maleic acid, the carrier being encapsulated by a polymer of monomers.

According to another aspect of the present invention, a polymer material having a carboxyl group or an amino group is swollen in an alkaline or acidic aqueous solvent, and after introducing a magnetic substance or a labeling substance into the swollen polymer material. And a method for producing an embedded polymer carrier comprising a step of treating with an acidic or alkaline aqueous solvent.

According to a preferred embodiment of the present invention, the above-mentioned method, wherein the polymer material having a carboxyl group or an amino group is a material comprising a synthetic polymer, wherein the synthetic polymer is polyacrylic acid or acrylic acid-styrene copolymer. The above method of coalescence; the pH of the alkaline aqueous solvent is 10 to 12;
The above method, wherein the acidic aqueous solvent has a pH of 3 to
5. The above-mentioned method which is an aqueous solvent of No. 5; the above-mentioned method in which the magnetic substance or the label is introduced simultaneously with the polymerization of the bifunctional monomer and / or the polyfunctional monomer; The above method, wherein the polymerization is carried out in the presence of an initiator; the above method, wherein the polymerization initiator is potassium persulfate.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION In the present specification, "embedding" does not mean that a substance such as a magnetic substance or a labeling substance is attached to the surface layer of a polymer material, but is coated with a polymer or the like and is blocked from the outside. It does not mean that the substance is embedded in the surface layer of the polymer material and is embedded without being blocked from the outside. "Swelling" refers to development in which when a cross-linked polymer material (solid) is immersed in a solvent (liquid), it absorbs the liquid and increases its volume to change the density of the polymer material. The “encapsulation” means a state in which a polymer having a small molecular weight such as an oligomer adheres to the surface layer of the polymer material, and the polymer material is wrapped by the attached polymer.

Hereinafter, the polymer carrier of the present invention and the method for producing the same will be described in more detail. As described above, the polymer carrier of the present invention is an embedded polymer carrier in which a magnetic substance or a labeling substance is embedded inside the surface layer of a polymer material having a carboxyl group or an amino group.

In the polymer carrier of the present invention, the magnetic substance or the labeling substance is present inside the surface layer of the polymer material and embedded in the gaps of the molecules constituting the polymer material. The embedding site of the magnetic substance or the labeling substance to be embedded is not particularly limited, but is usually a site within 25% from the surface layer of the polymer material, preferably a site within 15% from the surface layer. Appropriate.

The amount of the magnetic substance or the labeling substance embedded in the polymer material is not particularly limited, and any distribution amount may be used as long as the function can be exhibited in the intended use. As a polymer material that can be used, a material made of a synthetic polymer material having a carboxyl group or an amino group and having a property of swelling in an alkaline or acidic aqueous solvent is used.

The synthetic high molecular substance having a carboxyl group or an amino group includes a polymer of a polymerization monomer having a carboxyl group or an amino group as a functional group, and a polymerization monomer having a carboxyl group or an amino group as a functional group. Copolymers with monomers are mentioned. The polymerization monomer having a carboxyl group or an amino group has a carboxyl group or an amino group as a functional group, these functional groups may be substituted with a lower alkyl group, and has a conjugated double bond, Any compound can be used as long as it can form a polymer, for example, the following formula (I):

[0015]

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Wherein R ¹ , R ² , R ³ and R ⁴ are each independently at least one of a group —COOR and a group —NH
R and a group -CONH ₂ (where R represents C _n H _{2n + 1} ,
n represents an integer of 0 to 5), and the other represents a hydrogen atom. ] The compound represented by this is mentioned.

The compound represented by the above formula (I) includes
For example, acrylic acid, acrylate, acrylamide, ethyleneimine, aziridine and the like can be mentioned.

The monomer for polymerization may be any compound having a conjugated double bond and capable of forming a polymer and having no carboxyl group or amino group, for example, the following formula (II):

[0019]

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Wherein R ⁴ , R ⁵ , R ⁶ , R ⁷ and R ⁸ are
Each independently represents a group having at least one conjugated double bond such as a phenyl group or a biphenyl group, a halogen atom or a hydrogen atom. ] The compound represented by this is mentioned.

Examples of the compound represented by the formula (II) include styrene, butadiene, ethylene, and vinylidene chloride. As a polymer of these monomers, the following formula (III):

[0022]

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Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁴ , R ⁵ ,
R ⁶ , R ⁷ and R ⁸ are as described above, and one of m and n may be 0. ], For example, polyacrylic acid, acrylic acid-styrene copolymer and the like.

The material comprising the above-mentioned polymer substance used for the polymer carrier of the present invention can be prepared by subjecting the above-mentioned monomer to a method known per se, for example, a bulk polymerization method, an emulsion polymerization method, a seed (seed) weight. It is prepared by a legal method, a leap-free polymerization method, or the like.

The shape of the polymer material having a carboxyl group or an amino group is not particularly limited, but a spherical material is preferably used. The average particle size of the polymer material is usually 0.01 to 10 μm, and 0.1 to 1 μm.
m is more preferable, and a polymer material having these shapes and particle diameters can be prepared by polymerizing the monomer by the above method.

The magnetic material embedded in the polymer material may be a metal, for example, iron, ferrite, nickel, cobalt, chrome, or a metal such as molybdenum, copper, vanadium, manganese, aluminum, titanium, zinc, etc. Alloys with any one or more metals; iron oxides, such as pure Fe ₃ O ₄ or γ-Fe ₂ O ₃ , or other oxides such as cobalt, manganese, zinc, A substance in which barium, a rare earth or the like is combined, or a mixture thereof; chromium dioxide; Examples of the “labeling substance” include rare earth chelates such as europium, samarium, and terbium, phycocyanin,
Phycopyriproteins such as phycoerythrin, fluorescein, tetramethylrhodamine, Texas Red, 7
Fluorescent substances such as -amino-4-methylcoumarin; chemiluminescent substances such as acridinium derivatives and isoluminol;
Enzymes such as alkaline phosphatase, peroxidase and the like.

The particle diameter of the magnetic substance or the labeling substance is usually 10 to 1000 °, preferably 10 to 500 °. Further, the polymer carrier of the present invention is preferably one that is encapsulated with a water-soluble polymer of a bifunctional monomer and / or a polyfunctional monomer. The bifunctional monomer and the polyfunctional monomer may be any compounds as long as they can be polymerized to form a water-soluble polymer.
Maleic acid, ethylene imine, bisacrylamide, methyl bisacrylate and the like can be mentioned. The molecular weight of the polymer of these monomers that encapsulates the polymer carrier is usually 500 to
20,000, preferably about 1,000 to 3,000 is appropriate.

The method for producing the polymer carrier of the present invention described in detail above is not particularly limited, and a magnetic substance or a labeling substance is embedded in the surface layer of a polymer material having a carboxyl group or an amino group. As long as it is obtained, any method may be used. Specifically, for example, it can be produced by the following swelling method.

The polymer carrier of the present invention is obtained by swelling a polymer material having a carboxyl group or an amino group in an alkaline or acidic aqueous solvent, and introducing a magnetic substance or a labeling substance into the swollen polymer material. Alternatively, it can be produced by treating with an alkaline aqueous solvent.
That is, when the polymer material has a carboxyl group, it may be first swollen in an alkaline aqueous solvent and then treated with an acidic aqueous solvent, and when the polymer material has an amino group, it may be first acidic aqueous solution. Swell in a solvent and then treat with an alkaline aqueous solvent.

The polymer material having a carboxyl group or an amino group used in this production method is not particularly limited as long as it has a property of swelling in an alkaline or acidic aqueous solvent. Among the above-mentioned synthetic high-molecular substances (polymers), a high-molecular substance having a property of swelling in an alkaline aqueous solvent, for example, a polymer having a block unit represented by the above formula (III) Polyacrylic acid, acrylic acid-
A material made of a polymer material such as a styrene copolymer and having the above-mentioned shape and particle size is preferable. The polymer substance is
According to a commonly used method known per se, it can be prepared and used by the above-mentioned method, or a commercially available product can be used.

Examples of the aqueous solvent having the property of swelling the above-mentioned polymer material (hereinafter, this may be abbreviated as “solvent 1”) include, for example, those having a pH of 10 to 12 when the polymer material has a carboxyl group. Water solvent, a mixed solvent of water and dioxane, a mixed solvent of water and alcohol, or a mixed solvent of water and acetone; when the polymer material has an amino group, an aqueous solvent of pH 2-3, a mixed solvent of water and dioxane, A mixed solvent of water and alcohol or a mixed solvent of water and acetone can be used. Examples of the aqueous solvent for treating the swollen polymer material (hereinafter, this may be abbreviated as “solvent 2”) include, for example, an aqueous solvent having a pH of 3 to 5 when the polymer material has a carboxyl group. ;
When the polymer material has an amino group, pH 10 to 12
Water solvent.

The pH of the solvent 1 used is such that the polymer material having a carboxyl group or an amino group has the functional group, thereby increasing the affinity for water and the molar number of the functional group and the dissociation constant. For example, when a polymer material having a carboxyl group is used as the polymer material, alkaline water having a pH of 11 is used as the solvent 1 and a solvent 2 for treating the polymer material after swelling is used.
It is preferable to use acidic water of pH 3.

The swelling may be achieved by adding the solvent 1 to the polymer material and irradiating ultrasonic waves as needed to disperse the polymer material in the solvent 1. The amount of solvent 1 added to the polymer material is
An appropriate amount is about 100 to 1000 times the amount of 1 part by weight of the polymer material. The dispersion time until the swelling is completed is determined by the temperature of the solvent 1, and the higher the temperature, the shorter the swelling is completed. The dispersion time is usually from 0.5 to 60 minutes, preferably from 1 to 5 minutes, and the temperature of the solvent 1 is usually from a low temperature of 5 to 10 ° C to the boiling point of the solvent, preferably from 80 to 120 ° C. It is.

A magnetic substance or a labeling substance is added to the polymer material swollen as described above, and the mixture is stirred. The amount of the magnetic substance or the labeling substance to be added depends on the amount of about 80% of the total weight of the polymer material to be embedded in the case of the magnetic substance and the size thereof in the case of the labeling substance. It is appropriate that the size is 10 ° and about 200 to 300%. The stirring time of the mixture is usually 30 to 180 minutes, preferably 60 to 12 minutes.
0 minutes, the temperature is usually 40 to 100 ° C., preferably 50 to 100 ° C.
60 ° C. is suitable.

At this time, preferably, the above-mentioned bifunctional monomer and / or polyfunctional monomer is added, and if necessary, a polymerization initiator is added, and the monomers are reacted to simultaneously form a water-soluble polymer. . The reaction may be carried out under irradiation of ultrasonic waves as necessary, and the irradiation time of ultrasonic waves is suitably 30 seconds to 10 minutes, preferably 1 to 3 minutes.

As the polymerization initiator to be used, potassium persulfate, ammonium persulfate, the following formula (IV):

[0037]

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[0038] In the ^{formula, R 9, R 10, R} 11, R 12, R 13 and R ¹⁴ are each independently at least one of which hydrophilic group -N ⁺ H ₃ Cl ^- indicates, others Indicates a hydrogen atom. ]
And the like.

The type of the polymerization initiator is appropriately determined depending on the type of the bifunctional and / or polyfunctional monomer used. For example, when maleic acid is used as the bifunctional monomer, it is preferable to use potassium persulfate as the polymerization initiator. The amount of the polymerization initiator to be used is generally 0.001 to 0.1 mol, preferably 0.1 mol, per 1 mol of the monomer.
005-0.01 mol is suitable.

Next, a solvent 2, for example, pH, is added to the reaction mixture.
5.5 Add acidic water and shake for 8 hours to 24 hours to remove the solvent 1 from the polymer material. Solvent 2 for reaction mixture
Although the use amount of the solvent is not particularly limited, the solvent 2 is usually used in an amount of 10 to 100 times, preferably 100 times the volume of the reaction mixture.
It is appropriate to add up to 200 times the amount. The temperature of the solvent 2 is usually 0 to 10 ° C, preferably 0 to 5 ° C.

The treatment with the solvent 2 is carried out with shaking if necessary, and the treatment time is usually from 10 to 36 hours.
Further, the reaction mixture treated with the solvent 2 is subjected to an operation such as centrifugation to obtain a solid pellet containing a polymer carrier.
The pellets of the present invention are washed by using a known method which is known per se, using an acidic water and / or a phosphate buffer (about 0.1 to 0.5 M) having a pH of about 3 to 6 or the like. An embedded polymer carrier can be obtained.

In the polymer carrier thus obtained, a magnetic substance or a labeling substance is entangled in the gap of the surface polymer formed by swelling of the polymer material, and the polymer material shrinks by the treatment with the solvent 2, and the magnetic substance or the labeling substance becomes high. It has a state of being taken in the surface layer of the molecular material, that is, a structure that is embedded and distributed. Furthermore, by performing the polymerization of the bifunctional monomer and / or the polyfunctional monomer simultaneously with the addition of a magnetic substance or a labeling substance as necessary, a structure in which the polymer material is encapsulated by the polymer of the monomer is obtained. . The embedding position of the magnetic substance or the labeling substance in the polymer carrier is within 25% from the surface layer.

The embedded polymer carrier of the present invention, in which a magnetic substance or a label is embedded in the surface layer of a polymer material, has a uniform particle without dissociation of the magnetic substance or the label, and has good It has excellent properties such as dispersibility and can be used for various applications. For example, a polymer carrier in which a magnetic material is embedded can be used as a recording medium such as a magnetic tape, a separating agent for analysis, or a CD.
-It can be used as a coating material for ROM, a carrier for a DNA sequencer, and the like.

The polymer carrier of the present invention can be widely used in the field of diagnostics, particularly in immunological analysis as a support for immunologically active substances.

[0045]

The present invention will now be described more specifically with reference to examples. However, the following examples are to be regarded as helping to obtain a specific recognition of the present invention, and do not limit the scope of the present invention in any way.

Example 1 10% carboxyl group-modified latex (acrylic acid-
Styrene copolymer; average particle diameter 0.5 μm, manufactured by Seradyne Co., Ltd. Hereinafter, abbreviated as “CML”) 0.5 m aqueous dispersion
1 centrifuged (19,000 rpm, 10 minutes, 4
C) to produce solid CML pellets. This CML
5 ml of alkaline water having a pH of 10 to 12 was added to the pellets, dispersed by ultrasonication (SONIFER CELL DISTRIPUTOR; manufactured by Smith Kline) for 5 minutes, and stirred for a day.

To an alkaline dispersion of CML, 40 mg of maleic acid and 20 mg of a magnetic substance (average particle diameter 0.03 μm, manganese-zinc-iron; manufactured by Sakai Chemical Co., Ltd.) were added, and potassium persulfate, a water-soluble polymerization initiator, was added. (KPS) 7 mg was added. Ultrasonic wave (SONIFER CELL DISTRIP
UTOR; manufactured by Smith Kline) for 10 minutes to react. After the reaction, add alkaline water (pH
11) 40 ml was added and shaken all day and night.

The dispersion was centrifuged (19,000).
rpm, 10 minutes, 4 ° C.) to produce solid pellets. This pellet was dispersed in 20 ml of acidic water (pH 3), shaken all day and night to perform PS, and centrifuged again (1900, 1900).
0 rpm, 10 minutes, 4 ° C.) A solid pellet was prepared. The pellet is added to a 0.15 M phosphate buffer (pH 7).
Dispersed in 20 ml and shaken for 8 hours to obtain magnetic CML.

When the magnetic CML was observed with a scanning electron microscope, it was confirmed that the magnetic substance was uniformly present inside the surface layer of the CML. This magnetic CML aqueous dispersion was placed in a cuvette [quartz glass (h × w × d = 50 × 10 × 2 (m
m)], and a magnet was placed outside the cuvette to confirm the separability. Within 1 minute, all the magnetic CML was drawn to the inner wall of the cuvette, and the cuvette became transparent.

Claims (17)

[Claims] 1. An embedded polymer carrier comprising a magnetic substance or a labeling substance embedded in a surface layer of a polymer material having a carboxyl group or an amino group. 2. The polymer carrier according to claim 1, wherein the polymer material is a material made of a synthetic polymer substance. 3. The polymer carrier according to claim 2, wherein the synthetic polymer substance is polyacrylic acid or acrylic acid-styrene copolymer. 4. The polymer carrier according to claim 1, wherein the polymer material is spherical. 5. The spherical body has a particle size of 0.01 μm to 10 μm.
5. The polymer carrier according to claim 4, wherein 6. The magnetic material or the labeling substance is embedded within a range of 25% from the surface layer of the polymer material.
The polymer carrier according to any one of the above. 7. The method according to claim 7, wherein the polymer carrier comprises a bifunctional monomer and / or
The polymer carrier according to any one of claims 1 to 6, which is encapsulated by a polymer of a polyfunctional monomer. 8. The polymer carrier according to claim 7, wherein the bifunctional monomer is maleic acid. 9. A polymer material having a carboxyl group or an amino group is swollen in an alkaline or acidic aqueous solvent, a magnetic substance or a labeling substance is introduced into the swollen polymer material, and then the mixture is treated with an acidic or alkaline aqueous solvent. A method for producing an embedded polymer carrier, characterized by performing a treatment. 10. The method according to claim 9, wherein the polymer material is a material composed of a synthetic polymer substance. 11. The production method according to claim 10, wherein the synthetic polymer substance is a material composed of polyacrylic acid or acrylic acid-styrene copolymer. 12. An alkaline aqueous solvent having a pH of 10 to 10.
The production method according to any one of claims 9 to 11, which is an aqueous solvent of No. 12. 13. The production method according to claim 9, wherein the acidic aqueous solvent is an aqueous solvent having a pH of 3 to 5. 14. The production method according to claim 9, wherein the introduction of the magnetic substance or the labeling substance is carried out simultaneously with the polymerization of the bifunctional monomer and / or the polyfunctional monomer. 15. The method according to claim 14, wherein the bifunctional monomer is maleic acid. 16. The method according to claim 14, wherein the polymerization of the bifunctional monomer and / or the polyfunctional monomer is performed in the presence of a polymerization initiator. 17. The method according to claim 16, wherein the polymerization initiator is potassium persulfate.