CN103483626B - A kind of Magnetic Agarose sugar microballoon - Google Patents
A kind of Magnetic Agarose sugar microballoon Download PDFInfo
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- CN103483626B CN103483626B CN201310420312.3A CN201310420312A CN103483626B CN 103483626 B CN103483626 B CN 103483626B CN 201310420312 A CN201310420312 A CN 201310420312A CN 103483626 B CN103483626 B CN 103483626B
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- 229920000936 Agarose Polymers 0.000 title claims abstract description 57
- 239000002245 particle Substances 0.000 claims abstract description 32
- 239000000126 substance Substances 0.000 claims abstract description 15
- 239000011325 microbead Substances 0.000 claims abstract description 13
- 238000009736 wetting Methods 0.000 claims abstract description 8
- 230000005415 magnetization Effects 0.000 claims abstract description 7
- 238000005253 cladding Methods 0.000 claims abstract description 4
- 229910004298 SiO 2 Inorganic materials 0.000 claims description 10
- 229920000642 polymer Polymers 0.000 claims description 5
- 230000015572 biosynthetic process Effects 0.000 claims 1
- 239000002253 acid Substances 0.000 abstract description 5
- 230000005389 magnetism Effects 0.000 abstract description 4
- 239000002904 solvent Substances 0.000 abstract description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 29
- 238000000034 method Methods 0.000 description 7
- 238000003756 stirring Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 229920002189 poly(glycerol 1-O-monomethacrylate) polymer Polymers 0.000 description 3
- 229920002454 poly(glycidyl methacrylate) polymer Polymers 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 229910052701 rubidium Inorganic materials 0.000 description 3
- IGLNJRXAVVLDKE-UHFFFAOYSA-N rubidium atom Chemical compound [Rb] IGLNJRXAVVLDKE-UHFFFAOYSA-N 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 229910001220 stainless steel Inorganic materials 0.000 description 3
- 239000010935 stainless steel Substances 0.000 description 3
- 229920002818 (Hydroxyethyl)methacrylate Polymers 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000012501 chromatography medium Substances 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 238000012423 maintenance Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000004005 microsphere Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000007711 solidification Methods 0.000 description 2
- 230000008023 solidification Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
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- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
- Hard Magnetic Materials (AREA)
Abstract
Present invention is disclosed a kind of novel magnetic agarose microbeads, compared with other Magnetic Agarose sugar microballoon, it has following features: first, adopts the superparamagnetism Fe with high saturation and magnetic intensity
3o
4particle, as magnetic substance, ensures that it has high magnetic response performance and superparamagnetism; Secondly, at superparamagnetism Fe
3o
4the shell that particle outer cladding has unreactiveness, wetting ability and biocompatibility forms the magnetic core of core/shell structure, and then is filled in agarose microbeads by magnetic core and obtains Magnetic Agarose sugar microballoon.The invention solves Magnetic Agarose sugar microballoon magnetic response poor performance or easily in magnetic field magnetization produce the problem of remanent magnetism, by Fe
3o
4particles coat has the shell of unreactiveness, wetting ability and biocompatibility, makes Fe
3o
4particle acid and alkali-resistance, scale resistance and other solvent nature resistance to significantly improve, and ensure that Magnetic Agarose sugar microballoon has good biocompatibility, greatly improve practical application performance.
Description
Technical field
The present invention relates to a kind of novel magnetic agarose microbeads, belong to technical field of biological material, especially the separation and purification field of biomolecules.
Background technology
Magnetic polysaccharide microsphere is increasingly extensive in field application such as biological, chemistry, pharmacy and medical science in recent years, and it mainly applies the separation and purification, targeted drug, immunodetection, cellular segregation, chemical analysis etc. that comprise the biomolecules such as albumen, antibody, enzyme.Sepharose is natural polysaecharides chromatography media, it has the numerous characteristics of perfect medium, such as: high-hydrophilic, porousness, containing more can activated hydroxyl groups, not with biomacromolecule generation non-specific adsorption etc., be most widely used a kind of chromatography media up to now.Due on agarose containing more can activated hydroxyl groups, the medium of different aglucons as affinity chromatography, hydrophobic and ion-exchange chromatography can be accessed under certain conditions.Its separate object relates to the water-soluble biochemical substances such as protein, nucleic acid, peptide class, carbohydrate, and therefore Magnetic Agarose sugar microballoon is a kind of magnetic microsphere with wide application prospect.
Existing Magnetic Agarose sugar microballoon, mainly by inverse suspension method, by nanometer Fe
3o
4, that nanometer Fe, the magnetic substance such as micron order stainless steel particle and rubidium ferroboron particle are embedded in agarose is inner and obtain.There is following deficiency in actual use:
(1) because agarose shell is porous, with nanometer Fe
3o
4with nanometer Fe as magnetic substance be embedded in agarose formed Magnetic Agarose sugar microballoon, due to nanometer Fe
3o
4little with nanometer Fe particle diameter, surface-area is large, under extraneous acidic conditions easily and acid-respons cause iron to leak, magnetic is caused to decline, contact easily oxidized with oxidizing substance in preservation with use procedure simultaneously and cause use properties to decline, therefore its acid resistance and scale resistance poor, limit its practical application;
(2) to be embedded in agarose as magnetic substance with micron order stainless steel particle and rubidium ferroboron particle and to form Magnetic Agarose sugar microballoon, although acid and alkali-resistance and scale resistance increase, but because grain diameter is large, density is large, is very easy to sedimentation, is mainly used in expanded bed field, range of application is narrow, and easily magnetize in magnetic field and there is remanent magnetism, causing easily magnetic reunion between Magnetic Agarose sugar microballoon, not easily disperse;
(3) when being applied to the field such as biology, medical science, if adopt the direct embedded nano Fe of agarose
3o
4, nanometer Fe, the magnetic substance such as micron-sized stainless steel particle and rubidium ferroboron particle, because the magnetic substance biocompatibility of the inside embedding is poor, in use, biomolecules can be entered microballoon inside and directly contact with the magnetic substance of embedding by the duct on agarose shell, thus causes some biomolecules inactivation or cause stronger non-specific adsorption.
Summary of the invention
The object of this invention is to provide a kind of novel magnetic agarose microbeads solved the problems of the technologies described above, have acidproof, resistance to oxidation, the features such as good biocompatibility, high saturation and magnetic intensity and superparamagnetism.The present invention is reached by following technical proposals:
A kind of novel magnetic agarose microbeads, to have the superparamagnetism Fe of high saturation and magnetic intensity
3o
4particle as magnetic substance, at described superparamagnetism Fe
3o
4the shell that particle outer cladding has unreactiveness, wetting ability and biocompatibility forms the magnetic core of core/shell structure, and then is filled in agarose microbeads by magnetic core and forms Magnetic Agarose sugar microballoon.
The present invention further, described Fe
3o
4the saturation magnetization of particle is greater than 50emu/g, and has superparamagnetism.
Further, described shell comprises inorganics shell or high molecular polymer shell in the present invention, and preferably, described inorganics shell comprises SiO
2, described high molecular polymer shell comprises poly (glycidyl methacrylate), polystyrene-methacrylic acid glycidyl ester or poly-hydroxyethyl methacrylate.
The present invention further, described superparamagnetism Fe
3o
4mass ratio 1:1 ~ the 9:1 of particle and described shell, agarose is 1:1 ~ 10:1 with the magnetic nuclear mass ratio of core/shell structure.
Further, the particle diameter of the magnetic core of described core/shell structure is between 0.3 μm ~ 2 μm, and its saturation magnetization is greater than 55emu/g in the present invention.
Further, the particle diameter of described novel magnetic agarose microbeads is between 20 μm ~ 165 μm in the present invention.
Application of the present invention is implemented, and its significant technique effect is embodied in:
1. the present invention adopts the superparamagnetism Fe with high saturation and magnetic intensity
3o
4particle, as magnetic substance, improves Magnetic Agarose sugar microballoon magnetic response poor performance prepared by original technology or the problem of easily magnetizing generation remanent magnetism in magnetic field;
2. pass through in magnetic Fe
3o
4surface is introduced one deck and is had the good shell of unreactiveness, wetting ability and biocompatibility, Magnetic Agarose sugar acid and alkali-resistance, scale resistance and other solvent nature resistance to are significantly improved, improve the biocompatibility of magnetic core simultaneously, ensure that whole Magnetic Agarose sugar microballoon all has good biocompatibility, have a good application prospect.
Accompanying drawing explanation
Fig. 1 is the structural representation of novel magnetic agarose microbeads of the present invention;
Fig. 2 is the microphotograph figure of novel magnetic agarose microbeads of the present invention.
Embodiment
Below in conjunction with accompanying drawing and specific embodiment, the present invention will be described, for embodiment be only make generality to product of the present invention or method to illustrate, contribute to understanding the present invention better, but can't the scope of the invention be limited.Experimental technique described in following embodiment, if no special instructions, is ordinary method, described material, if no special instructions, all can obtain from commercial channels.
The present invention relates to a kind of novel magnetic agarose microbeads, to have the superparamagnetism Fe of high saturation and magnetic intensity
3o
4particle, as magnetic substance, makes the Magnetic Agarose sugar microballoon finally prepared have high saturation and magnetic intensity and superparamagnetism, can ensure that it has the magnetic response time be exceedingly fast like this, can ensure that again it is not magnetized and produces remanent magnetism in magnetic field.
As shown in Figure 1, be the structural representation of novel magnetic agarose microbeads of the present invention, described Fe
3o
4/ SiO
2the saturation magnetization of magnetic core is greater than 70emu/g, has superparamagnetism, at described superparamagnetism Fe
3o
4particle outer cladding has the shell of unreactiveness, wetting ability and biocompatibility and forms magnetic core, and the material of described shell comprises inorganics or high molecular polymer.Preferably, described inorganics includes but not limited to SiO
2, described high molecular polymer includes but not limited to poly (glycidyl methacrylate), polystyrene-methacrylic acid glycidyl ester or poly-hydroxyethyl methacrylate.
Above-mentioned shell can effectively protect magnetic substance coated in it, improves the Magnetic Agarose sugar resistance to acids and bases of microballoon, scale resistance and tolerates the performance of other solvent; Iron in use procedure is avoided to leak or cause because of oxidation magnetic to reduce; Simultaneously because shell has wetting ability, can be well compatible with the wetting ability agarose layer of subsequent packet quilt, be beneficial to magnetic substance dispersed within it, as shown in Figure 2; Shell also has good biocompatibility in addition, makes the present invention in subsequent bio molecular separation purge process, can not make biomolecules inactivation, also can not cause non-specific adsorption.
Also be coated with agarose layer outward at described magnetic core, agarose is 1:1 ~ 10:1, described superparamagnetism Fe with the magnetic nuclear mass ratio of core/shell structure
3o
4mass ratio 1:1 ~ the 9:1 of particle and described shell.Particularly, the particle diameter of the magnetic core of described core/shell structure is between 0.3 μm ~ 2 μm, and its saturation magnetization is greater than 55emu/g.By upper, the particle diameter of the novel magnetic agarose microbeads that the present invention finally obtains is between 20 μm ~ 165 μm.
[embodiment 1]
With Fe
3o
4/ SiO
2preparation for the Magnetic Agarose sugar microballoon of magnetic core:
Measure 400mL hexanaphthene with 500mL graduated cylinder to add in 1000mL there-necked flask, and then add class of 4mL department 85, there-necked flask is placed in 60 DEG C of waters bath with thermostatic control, motor machine stirs 30min.
Take 6.0g agar Icing Sugar with analytical balance to add in 500mL Erlenmeyer flask, then add 100mLFe
3o
4/ SiO
2content is the aqueous solution of 5mg/mL, by above-mentioned solution heating for dissolving; The stirring velocity adjusted in above-mentioned hexanaphthene oil phase is 1600rpm, contains Fe by what prepare
3o
4/ SiO
2agarose solution slowly add in hexanaphthene oil phase, the rotating speed adding rear maintenance 1600rpm stirs 60min, finally stop heating in water bath, hot water in water-bath is extracted out, adding frozen water cooling 10min makes microballoon solidify, and the microballoon after solidification obtains the Magnetic Agarose sugar microballoon that particle diameter is 20 μm ~ 165 μm after washing, purifying.
[embodiment 2]
With Fe
3o
4/ PGMA(poly (glycidyl methacrylate)) for magnetic core Magnetic Agarose sugar microballoon preparation:
The mode Fe of reference example 1
3o
4/ PGMA replaces Fe
3o
4/ SiO
2endorse as magnetic and obtain the embedding Fe that particle diameter is 20 μm ~ 165 μm
3o
4the Magnetic Agarose sugar microballoon of/PGMA.
[embodiment 3]
With Fe
3o
4/ SiO
2preparation for the Magnetic Agarose sugar microballoon of magnetic core:
Measure 400mL hexanaphthene with 500mL graduated cylinder to add in 1000mL there-necked flask, and then add class of 8mL department 85, there-necked flask is placed in 70 DEG C of waters bath with thermostatic control, motor machine stirs 30min.
Take 6.0g agar Icing Sugar with analytical balance to add in 500mL Erlenmeyer flask, then add 100mLFe
3o
4/ SiO
2content is the aqueous solution of 5mg/mL, heating for dissolving; The stirring velocity finally adjusted in above-mentioned hexanaphthene oil phase is 2000rpm, the agarose solution prepared slowly is added in oil phase, the rotating speed adding rear maintenance 2000rpm stirs 60min, finally stop heating in water bath, hot water in water-bath is extracted out, adding frozen water cooling 10min makes microballoon solidify, and the microballoon after solidification obtains the Magnetic Agarose sugar microballoon that particle diameter is 20 μm ~ 75 μm after washing, purifying.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the technology of the present invention principle; can also make some improvement and modification, these improve and modification also should be considered as protection scope of the present invention.
Claims (6)
1. a Magnetic Agarose sugar microballoon, is characterized in that:
To have the superparamagnetism Fe of high saturation and magnetic intensity
3o
4particle as magnetic substance, at described superparamagnetism Fe
3o
4particle outer cladding has the magnetic core of the shell formation core/shell structure of unreactiveness, wetting ability and biocompatibility, and then magnetic core is filled in agarose microbeads forms Magnetic Agarose sugar microballoon, the particle diameter of the magnetic core of described core/shell structure between 0.3 μm ~ 2 μm, described in there is the superparamagnetism Fe of high saturation and magnetic intensity
3o
4the saturation magnetization of particle is greater than 70emu/g.
2. a kind of Magnetic Agarose sugar microballoon according to claim 1, is characterized in that: described shell comprises inorganics shell or high molecular polymer shell.
3. a kind of Magnetic Agarose sugar microballoon according to claim 2, is characterized in that: described inorganics shell comprises SiO
2layer.
4. a kind of Magnetic Agarose sugar microballoon according to claim 1, is characterized in that: described superparamagnetism Fe
3o
4mass ratio 1:1 ~ the 9:1 of particle and described shell, described agarose is 1:1 ~ 10:1 with the magnetic nuclear mass ratio of described core/shell structure.
5. a kind of Magnetic Agarose sugar microballoon according to claim 1, is characterized in that: the saturation magnetization of the magnetic core of described core/shell structure is greater than 55emu/g.
6. a kind of Magnetic Agarose sugar microballoon according to claim 1, is characterized in that: the particle diameter of described Magnetic Agarose sugar microballoon is between 20 μm ~ 165 μm.
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| CN110177617A (en) * | 2017-01-04 | 2019-08-27 | 南京金斯瑞生物科技有限公司 | The alkaline-resisting albumin A magnetic bead of high carrying capacity and its application method |
| WO2018137633A1 (en) * | 2017-01-24 | 2018-08-02 | 珠海神平医疗科技有限公司 | Liquid embolic material and preparation method therefor |
| CN107042093B (en) * | 2017-03-07 | 2019-07-12 | 四川大学 | A kind of preparation method of the big ball adsorbent material of composite magnetic hydroxyapatite |
| CN114570302B (en) * | 2022-03-24 | 2024-08-09 | 苏州华诺生物科技有限公司 | Magnetic agarose porous microsphere based on protein and preparation method thereof |
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| CN103212377B (en) * | 2013-04-19 | 2014-09-24 | 哈尔滨益材新材料有限公司 | A kind of preparation method and application of agarose immunomagnetic microspheres |
| CN103223322B (en) * | 2013-05-13 | 2015-02-04 | 洛阳师范学院 | Preparation method of nanosilver and sulfydryl jointly modified magnetic microspheres |
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Address after: Xinghu Street Industrial Park of Suzhou city in Jiangsu province 215123 No. 218 BioBAY A6-101 Patentee after: Suzhou beaver Biomedical Engineering Co., Ltd. Address before: Xinghu Street Industrial Park of Suzhou city in Jiangsu province 215123 No. 218 BioBAY A6-101 Patentee before: Beaver Nano-Technologies Co., Ltd. |