CN104538168A - Magnetic bead preparing method and application - Google Patents
Magnetic bead preparing method and application Download PDFInfo
- Publication number
- CN104538168A CN104538168A CN201410802284.6A CN201410802284A CN104538168A CN 104538168 A CN104538168 A CN 104538168A CN 201410802284 A CN201410802284 A CN 201410802284A CN 104538168 A CN104538168 A CN 104538168A
- Authority
- CN
- China
- Prior art keywords
- magnetic
- magnetic bead
- particle
- preparation
- nano
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a magnetic bead preparing method and application, and belongs to the technical field of magnetic material preparing. The preparing method comprises the steps that firstly, magnetic nanoparticles with the surface functionalization are prepared, then, at least two magnetic nanoparticles with the surface functionalization are connected through a cross-linking agent, and a magnetic bead with the magnetic content of 70 %-98 % is prepared. The problems that the magnetic content of an existing magnetic bead is low, to achieve fast separation, the size of the magnetic bead is greatly increased, and consequently the suspension stability is reduced are solved. In addition, for the captured component biological reaction, particularly for the immunoreaction, the steric hindrance caused by the small magnetic bead is obviously reduced, the capturing capability and the analyzing and detecting sensitivity are improved, the magnetic bead can be better applied to the biological medicine separation and analysis, and the magnetic bead is particularly suitable for separation of cells, bacteria, viruses, DNA/RNA, protein and the like in biological body fluid or reaction fluid and the clinical biochemistry fast automatic detection based on chemiluminiscence electrochemical luminescence and fluorescence.
Description
Technical field
The invention belongs to magnetic material preparing technical field, particularly relate to a kind of preparation method and application of magnetic bead.
Background technology
For the magnetic nano-particle of conventional method synthesis, usually show the notable feature of following two aspects.First, show superparamagnetism or approximate superparamagnetism (remanent magnetism is less or few), be conducive to building up from the solution of dispersion under magnetic fields and being separated, also be again distributed in solution after being conducive to cancelling magnetic fields simultaneously, and the analysis realized separation component, above-mentioned feature is particularly conducive to automation separation and analysis; The second, particle diameter (referring to the non-hydrated particle diameter characterized with TEM) is less, usually at 10-100nm, each particle is under strong magnetic field action, show less magnetic force, from the solution of dispersion disengaging time long, be separated not exclusively, thus cause that analysis speed is slow, sensitivity for analysis reduces.
In order to utilize above-mentioned advantage (superparamagnetism or approximate superparamagnetism) and overcome above-mentioned shortcoming (disengaging time long, is separated incomplete), multiple magnetic nano-particle must be assembled into the larger magnetic bead of particle diameter (in order to the convenience of following expression, the particle of that namely magnetic nano-particle refers to not connect, small particle diameter, nanoscale, namely magnetic bead refers to particle after connecting, Large stone).The magnetic force of a magnetic bead namely added by the magnetic force of multiple magnetic nano particle and, make the magnetic force of magnetic bead much larger than magnetic nano-particle, can gather fast under magnetic fields thus be separated fast, completely in the solution of dispersion.Meanwhile, because magnetic bead is assembled by magnetic nano-particle, namely remain the superparamagnetism of magnetic nano-particle or approximate superparamagnetism, the rapid dispersion again of magnetic bead can be realized after cancelling magnetic fields.For the magnetic bead that bio-separation uses with analysis, in fact also has another important requirement, good suspension stability must be had in the solution, otherwise can not carry out within certain reaction time and under suspended state fast with separated component, complete reaction.Therefore, for bio-separation and the condition analyzing the magnetic bead that uses and must possess three aspects of a performance brilliance, one, strong magnetic force, preferably in 1-2 minute under the effect of conventional strong magnet (ndfeb magnet), realize dispersion magnetic bead in the solution quick, be separated completely; Its two, superparamagnetism or approximate superparamagnetism, make magnetic bead (as jolting, liquid flow) rapid dispersion again under more weak External Force Acting; Its three, good suspension stability, in aqueous can be suspending stabilized at least 30 minutes, sedimentation do not occur.
For existing magnetic bead, usually between strong magnetic force and suspension stability, there is contradiction.In order to obtain strong magnetic force, must improve the quantity of magnetic nano-particle in magnetic bead, but the volume of magnetic bead often increasing fast, causing suspension stability to reduce, also cause magnetic bead to the sterically hindered increase of catching object simultaneously, capture ability reduces.The principal element that magnetic bead volume increases fast is gathering of magnetic nano particle on the one hand, is build up for the chemistry of the non-magnetic components of immobilized magnetic nano particle on the other hand.The former can not avoid, and therefore, namely unique way that solution magnetic bead increases fast is reduce the content of the latter in magnetic bead, namely improves the magnetic content (magnetic content is the content of magnetisable material in magnetic bead (magnetic nano particle)) of magnetic bead.Existing commercialization magnetic bead, comprises Medical magnetic ball, and great majority are all high molecular polymer magnetic beads, comprises homopolymers and copolymer magnetic bead.Conventional synthesis macromolecule comprises polystyrene, polyacrylic acid, polymethylacrylic acid, polyethylene glycol, PVP, PLA, polyvinyl alcohol and their copolymer etc.Conventional preparation method comprises investment, dispersion copolymerization method, emulsion polymerization and living polymerization etc.These methods are all in the process of polymerizable organic monomer polymerization reaction take place, are coated among polymer microsphere by magnetic nano particle, thus make immobilized multiple magnetic nano particle in a macromolecule magnetic bead.In order to make magnetic nano particle stably immobilized, necessarily require each magnetic nano particle in magnetic bead all will have enough polymeric PTC materials layers, so namely result in the increase of namagnetic substance content in magnetic bead, namely magnetic content is lower.Except macromolecule magnetic bead, in fact, organic and/or inorganic materials magnetic bead is gone back.The method of hydrolysis organosiloxane is adopted to prepare SiO
2coated magnetic bead, preparation method mainly comprises reverse microemulsion method, aerosol high-temperature decomposition and sol-gal process etc.Under the prerequisite ensureing magnetic force, the magnetic content of this kind of magnetic bead is lower.Above-mentioned two class magnetic beads, all by coated (comprise physics coated and based on immobilized coated of chemical bond) immobilized magnetic nano particle in the process preparing non-magnetic microspheres, this is the content inevitably increasing namagnetic substance, thus reduce the magnetic content preparing magnetic bead, this is inherent intrinsic contradictions, is difficult to avoid from method.
Summary of the invention
The object of the invention is the preparation method proposing a kind of magnetic bead, the chemical bond that this preparation method utilizes the chemical reaction between the reactive group of crosslinking agent and the surface functional group of magnetic nano-particle to be formed, multiple magnetic nano-particle is coupled together, prepares the magnetic bead of high magnetic content.
For reaching this object, the present invention by the following technical solutions:
Invention provides a kind of preparation method of magnetic bead, first prepares surface-functionalized magnetic nano-particle, then adopts crosslinking agent to be coupled together by least two surface-functionalized magnetic nano-particles, prepares the magnetic bead that magnetic content is 70%-98%.
As a kind of preferred version, the particle diameter of described surface-functionalized magnetic nano-particle is 5-250nm, and described surface-functionalized magnetic nano-particle is the magnetic nano-particle of finishing or embeds the immobilized macromolecular magnetic nano-particle of macromolecular magnetic nano-particle or surface.
As a kind of preferred version, the surface functional group of described surface-functionalized magnetic nano-particle is one or more in hydroxyl or carboxyl or amino or aldehyde radical or sulfydryl.
As a kind of preferred version, described crosslinking agent adopts the macromolecular substances containing reactive group, and described reactive group is one or more in hydroxyl or carboxyl or aldehyde radical or sulfydryl or amino.
As a kind of preferred version, described macromolecular substances is one or more in polymer or block polymer or protein or DNA or polypeptide or polysaccharide.
As a kind of preferred version, the particle diameter of described magnetic bead is at least 2 times of described magnetic nano-particle particle diameter, is specially 200nm-50 μm.
As a kind of preferred version, described magnetic bead is spherical or irregular shape.
As a kind of preferred version, described magnetic bead complete disengaging time is in the solution 1min-6min.
The magnetic bead that the preparation method that present invention also offers a kind of as above arbitrary described magnetic bead obtains is in biological medicine analysis and the application be separated, described magnetic bead is directly added in biological sample, utilizes charge interaction to carry out the magnetic fast Acquisition separation and analysis of object;
Or first coupling capture agent on described magnetic bead, then the magnetic fast Acquisition separation and analysis carrying out object.
As a kind of preferred version, described capture agent is antibody or fit.
Beneficial effect of the present invention is:
The invention provides a kind of preparation method of magnetic bead, the chemical bond that the present invention utilizes the chemical reaction between the reactive group of crosslinking agent and the surface functional group of magnetic nano-particle to be formed, multiple magnetic nano-particle is coupled together, prepares the magnetic bead of high magnetic content (70%-98%).Preparation method of the present invention is simple, and prepare productive rate high, magnetic bead particle size is controlled; Preparing the magnetic nano particle of immobilized identical amount in magnetic bead, namely under each magnetic bead has the prerequisite of identical magnetic force, the magnetic content of magnetic bead prepared by the present invention is higher, volume is less, and surface charge is enriched, and surface hydrophilicity is stronger, suspension stability is better, solving the existing magnetic bead problem that particularly Medical magnetic ball magnetic content is low, also overcoming existing magnetic bead to realize quick separating, greatly increase magnetic bead volume and the problem that causes suspension stability to reduce.
Described magnetic bead, in biological medicine analysis and the application be separated, directly adds in biological sample by the magnetic bead that the preparation method that present invention also offers a kind of above-mentioned magnetic bead obtains, and utilizes charge interaction to carry out the magnetic fast Acquisition separation and analysis of object; Or first coupling capture agent on described magnetic bead, then the magnetic fast Acquisition separation and analysis carrying out object.For the particularly immune response of captured component biological respinse, the sterically hindered remarkable reduction that the small size magnetic bead that the present invention obtains causes, capture ability and analyzing and testing sensitivity improve, can better for biological medicine separation and analysis, be specially adapted to the separation of cell, bacterium, virus, DNA/RNA, protein etc. in biological fluid or reactant liquor and the rapid automatized detection of clinical biochemical based on chemiluminescence, electrochemical luminescence and fluorescence.
Accompanying drawing explanation
Fig. 1 is the thermogravimetric curve figure that the PEI of the embodiment of the present invention one is cross-linked magnetic bead;
Fig. 2 is that the PEI of the embodiment of the present invention one is cross-linked magnetic bead and compares with certain popular Medical magnetic ball, the surplus ratio percentage of not separated magnetic bead (in the solution) curve chart over time of magnetic bead when Magneto separate;
Fig. 3 is the mould curve chart over time that the PEI of the embodiment of the present invention one is cross-linked the ultra-violet absorption spectrum of magnetic bead solution.
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is described in further detail.Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.
The technical problem solved for making the present invention, the technical scheme of employing and the technique effect that reaches are clearly, be described in further detail below in conjunction with the technical scheme of accompanying drawing to the embodiment of the present invention, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those skilled in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
As everyone knows, surface-functionalized magnetic nano-particle is mainly divided into following several:
1, the magnetic nano-particle of non-finishing: the magnetic nano-particle prepared by conventional method, comprises Fe
3o
4, Fe
2o
3and their heterozygote, CoFe
2o
4, NiFeO
4, ZnFe
2o
4, MnFeO
4deng, numerous functional groups on surface refer to a large amount of hydroxyls (-OH).
2, the magnetic nano-particle of finishing: for the magnetic nano-particle prepared by conventional method, change the functional group of particle surface by chemical modification, corresponding functional group can be hydroxyl or carboxyl or amino or aldehyde radical or sulfydryl wherein one or more.
3, embed macromolecular magnetic nano-particle: while preparing magnetic nano-particle by conventional method, add the large molecule of needs embedding, make its macromolecule fraction be embedded in inside particles, another part is outside at particle.But not getting rid of some large molecule is the surface being attracted to particle.The macromolecular reactive group of particle outside is surface functional group.
4, the immobilized macromolecular magnetic nano-particle in surface: after preparing magnetic nano-particle by conventional method, utilize one or more active forces in chemical bond, hydrophobic effect, charge effect by immobilized for large molecule in particle surface.But not getting rid of some large molecule is the surface being attracted to particle.The macromolecular reactive group of particle outside is surface functional group.
Embodiment one
Present embodiments provide a kind of preparation method of magnetic bead, comprise the following steps:
(1) the magnetic nano-particle preparation of citric acid modification: by the Fe of preparation
3o
4magnetic nano-particle aqueous-based dispersions ultrasonic disperse 15min, gets 20mL and is placed in logical nitrogen deoxygenation 20min in three-necked bottle, join ultrasonic 15min in a certain amount of sodium citrate solution, make Fe
3o
4magnetic nano-particle is uniformly dispersed in system, connect experimental provision, keep nitrogen environment, churned mechanically speed is adjusted to 600rpb, and stirring reaction 1h after water-bath temperature rises to 60 DEG C, carries out Magneto separate with ndfeb magnet, unnecessary natrium citricum is removed in tri-distilled water washing, can not obtain the supernatant of clarification until Magneto separate till, be finally again scattered in 20mL tri-distilled water, the water-soluble Fe of obtained citric acid modification
3o
4, i.e. the magnetic nano-particle of citric acid modification, surface functional group is carboxyl.
(2) PEI of magnetic nano-particle is crosslinked prepares magnetic bead: in the aqueous-based dispersions of 15mg magnetic nano-particle, adopt EDCHCl and sulfo-NHS activated carboxyl, shaking table activation 1h under the reaction condition of 37 DEG C, Magneto separate, wash three times, to be added in round-bottomed flask ultrasonic disperse 1min under 37 DEG C of water bath condition; Get polymer P EI (Mw=1800) aqueous solution of 12.5mg in round-bottomed flask, and the cushioning liquid adding pH=7.4 is placed on jolting in shaking table, reaction 1h, Magneto separate, washing, multiple soluble in water, obtain the magnetic bead that PEI is crosslinked.
The particle diameter of magnetic bead prepared by the present embodiment and Zeta potential analysis:
As can be seen from subordinate list 1, for the particle diameter of dynamic light scattering determination, ChiSquared value and the P.I. value of magnetic bead prepared by the present embodiment are all less, illustrate that domain size distribution is satisfied with normal distribution well, and are evenly distributed.Further, magnetic bead is with abundant surface charge, and its corresponding particle diameter is much smaller than the particle diameter (about 2700nm) of existing Medical magnetic ball.Above-mentioned feature makes it show good suspension stability in aqueous.What is more important, small scale magnetic bead will greatly reduce sterically hindered to captured component of magnetic bead, increases capture ability, improves detection sensitivity, reduces capture time, realizes rapid analysis and detects.
The particle diameter of magnetic bead prepared by the PEI showing 1Mw=1800 and Zeta potential
The thermogravimetric curve of magnetic bead prepared by the present embodiment and magnetic content analysis:
We have done thermal weight loss sign to magnetic bead prepared by PEI crosslinked (Mw=1800), as shown in Figure 1, as can be seen from thermogravimetric curve we, weightlessness is point two stages: the first stage be temperature below 140 DEG C time, the weightlessness of this section is the water evaporation in magnetic bead top layer, and now magnetic bead magnetic content is 99.12%; Second stage is from 200 DEG C, and this is because non-magnetic components (comprising surface-functionalized citric acid and large molecule PEI) contained in magnetic bead decomposes the weightlessness caused.Temperature is after 640 DEG C, and thermogravimetric curve tends towards stability, and is remaining Fe
3o
4.Based on the thermal weight loss rate comprising the non-magnetic components of moisture content, the magnetic content calculating magnetic bead is 91.87%.Certainly, the impact of removing moisture content, magnetic content can be higher, and this is very advantageous in the efficient magnetic bead preparation of strong magnetic force, small size, higher suspension stability.
Magnetic bead prepared by the present embodiment compares with the Magneto separate of Medical magnetic ball:
When Magneto separate, the percentage (the present invention is defined as " magnetic bead surplus ratio ") accounting for total magnetic bead with magnetic bead not separated in solution, for standard, investigates the Magneto separate ability being cross-linked magnetic bead and certain popular Medical magnetic ball with PEI.From accompanying drawing 2, when additional magnetic fields 1min, not separated surplus ratio, Medical magnetic ball is cross-linked magnetic bead a little less than the PEI of the present embodiment, but the surplus ratio that PEI is cross-linked magnetic bead after 2min is just less than the surplus ratio of Medical magnetic ball.PEI is cross-linked magnetic bead in 6min, achieves intimate 100% separation, but Medical magnetic ball all can not realize 100% separation for a long time, and its surplus is up to more than 10%, and this just imply that the separated component up to more than 10% can not realize being separated.For the high-sensitive luminesceence analysis (as widely used chemiluminescence, electrochemiluminescence analysis) in clinical examination, imply that a large amount of low content samples will occur false negative.This biomarker for present clinical widely used early diagnosis of cancer detects, and is extremely disadvantageous, very easily occurs undetected in a large number.Show thus, magnetic bead prepared by the present embodiment, Magneto separate is much better than certain popular Medical magnetic ball.
The suspension stability of magnetic bead prepared by the present embodiment and Medical magnetic ball compares:
Because the ultraviolet spectra of magnetic bead does not have characteristic absorption, with the mould of ultraviolet light absorption spectrum over time curve characterize the magnetic bead content in solution.Still with certain popular Medical magnetic ball for standard of comparison, from accompanying drawing 3, there is not significant difference in two class magnetic beads suspension stability in the solution in 20min, all shows good stability.But along with the increase of time, both difference is just extremely obvious, and when reaching 120nm, magnetic bead prepared by the present embodiment remains extremely stable, does not find obvious sedimentation, but the Medical magnetic ball sedimentation of contrast 15%.Show that magnetic bead prepared by the present embodiment has fabulous suspension stability, be much better than certain popular Medical magnetic ball.
Embodiment two:
The preparation method of a kind of magnetic bead that the present embodiment provides is substantially identical with the preparation method of the magnetic bead described in embodiment one, and difference part is:
(1) there is the preparation of the magnetic nano-particle of the finishing of Small molecular functional group: by the Small Molecule Functionalization reagent containing functional group (carboxyl contained by other carboxylic acid comprising hydroxyl, amino, aldehyde radical, sulfydryl and be different from citric acid), replace the citric acid of step (1) in embodiment one, carry out the finishing of similar magnetic nano-particle, the magnetic nano-particle of the finishing of difference in functionality group must be had.
(2) preparation of the immobilized macromolecular magnetic nano-particle in surface: by the large molecule needing surface immobilized, the block polymer, protein, DNA, polypeptide, polysaccharide etc. that comprise polymer macromolecule, connect with chemical bond replace the Small Molecule Functionalization reagent in (1) of the present embodiment, by similar method, obtain surperficial immobilized macromolecular magnetic nano-particle.
(3) preparation of the magnetic bead of macromolecules cross-linking: large molecule comprised polymer macromolecule, replace the polymer P EI of step (2) in embodiment one with block polymer, protein, DNA, polypeptide, polysaccharide etc. that chemical bond connects, the magnetic bead of different macromolecules cross-linking can be prepared according to similar method.
Embodiment three
The preparation method of a kind of magnetic bead that the present embodiment provides is substantially identical with the preparation method of the magnetic bead described in embodiment one, and difference part is:
(1) embed the magnetic nano-particle preparation of PEI: get the tri-distilled water of 80mL in the four-necked bottle of 250mL, logical nitrogen, except after oxygen 20min, connects experimental provision, checks air-tightness, ensures that air-tightness is good; Bath temperature is adjusted to 35 DEG C, churned mechanically rotating speed is adjusted to 300rpb, keeps the nitrogen environment in four-necked bottle, accurately takes the FeCl of 4.05g
36H
2the FeCl of O and 1.98g
24H
2o, join stirring and dissolving 20min in four-necked bottle, in certain density ammonia spirit, add a certain amount of polymer macromolecule PEI (Mw=10000), churned mechanically rotating speed is adjusted to 600rpb, slow dropping ammoniacal liquor dilution, until pH value of solution=11, stops dripping ammoniacal liquor; Reaction 1h is continued after bath temperature being risen to 60 DEG C.
After reaction terminates, be transferred in beaker, Magneto separate, abandon supernatant by the suspension of black, redissolve with tri-distilled water and wash, Magneto separate, abandons supernatant liquor, so repeats, until solution is in neutral; By the Fe obtained
3o
4nanoparticle is dispersed in ultrasonic 30min in the tri-distilled water of 100mL, leaves standstill and obtain the magnetic nano-particle of embedding PEI.
(2) PEI of magnetic nano-particle is crosslinked prepares magnetic bead: adopt the carboxyl in EDCHCl and sulfo-NHS activated polyacrylic acid, shaking table activation 1h under the condition of 37 DEG C, added ultrasonic disperse 1min in the aqueous dispersions of the magnetic nano-particle of a certain amount of embedding PEI, shaking table jolting reaction 1h is placed under 37 DEG C of water bath condition, Magneto separate, washing, multiple soluble in water, obtain magnetic bead.
In addition, the PEI that can the large molecule containing a large amount of reactive group (amino contained by other polymerization organic amine comprising hydroxyl, carboxylic acid, aldehyde radical, sulfydryl and be different from PEI) be replaced in (1) of the present embodiment, carry out similar macromolecular embedding, the macromolecular magnetic nano-particle of different embedding must be had.
Embodiment four
The magnetic bead that the preparation method present embodiments providing a kind of magnetic bead as described in embodiment one or embodiment two or embodiment three obtains is in biological medicine analysis and the application be separated, magnetic bead is directly added in biological sample, utilizes charge interaction to carry out the magnetic fast Acquisition separation and analysis of object; Or first coupling capture agent on magnetic bead, then the magnetic fast Acquisition separation and analysis carrying out object.
Now illustrate for the magnetic bead of the surface of embodiment three with carboxyl.By the magnetic EDC-NHS method activated carboxyl of 50 μ L 5mg/mL, Magneto separate, redissolve in the EP pipe of the low absorption of 1.5mL albumen, add the PBS of 200ul0.01M pH7.4, vortex makes it even, adds 50 μ L 40 μ g/mL rabbit anti-mouse IgG and hatches 30min, the BSA adding 1% of 50 μ L closes 30min, Magneto separate, the PBS adding 300 μ L 0.01M pH7.4 redissolves, and obtains the immunomagnetic beads of coupling capture agent rabbit anti-mouse IgG antibody.
For the particularly immune response of captured component biological respinse, the sterically hindered remarkable reduction that the small size magnetic bead that the present invention obtains causes, capture ability and analyzing and testing sensitivity improve, can better for biological medicine separation and analysis, be specially adapted to the separation of cell, bacterium, virus, DNA/RNA, protein etc. in biological fluid or reactant liquor and the rapid automatized detection of clinical biochemical based on chemiluminescence, electrochemical luminescence and fluorescence.
The particle size range of the magnetic nano-particle that the present invention is surface-functionalized is 5-250nm, and described surface-functionalized magnetic nano-particle for the magnetic nano-particle of finishing or can embed the immobilized macromolecular magnetic nano-particle of macromolecular magnetic nano-particle or surface.The particle diameter of magnetic bead is at least 2 times of magnetic nano-particle particle diameter, and concrete scope is 200nm-50 μm, and namely the hydration particle diameter of the magnetic bead of dynamic light scattering determination is more than 200nm, maximumly reaches 50 μm, and the form of magnetic bead is spherical or irregular shape.Magnetic bead complete disengaging time in the solution controls in 1min-6min scope, and namely the quick separating shortest time of magnetic bead is 1min.
The chemical bond that the present invention utilizes the chemical reaction between the reactive group of crosslinking agent and the surface functional group of at least two magnetic nano-particles to be formed, multiple magnetic nano-particle is coupled together, prepares the magnetic bead of high magnetic content (70%-98%).
Preparation method of the present invention is simple, and prepare productive rate high, magnetic bead particle size is controlled; Preparing the magnetic nano particle of immobilized identical amount in magnetic bead, namely under each magnetic bead has the prerequisite of identical magnetic force, the magnetic content of magnetic bead prepared by the present invention is higher, volume is less, and surface charge is enriched, and surface hydrophilicity is stronger, suspension stability is better, solving the existing magnetic bead problem that particularly Medical magnetic ball magnetic content is low, also overcoming existing magnetic bead to realize quick separating, greatly increase magnetic bead volume and the problem that causes suspension stability to reduce.
Below know-why of the present invention is described in conjunction with specific embodiments.These describe just in order to explain principle of the present invention, and can not be interpreted as limiting the scope of the invention by any way.Based on explanation herein, those skilled in the art does not need to pay performing creative labour can associate other embodiment of the present invention, and these modes all will fall within protection scope of the present invention.
Claims (10)
1. a preparation method for magnetic bead, is characterized in that: first prepare surface-functionalized magnetic nano-particle, then adopts crosslinking agent to be coupled together by least two surface-functionalized magnetic nano-particles, prepares the magnetic bead that magnetic content is 70%-98%.
2. the preparation method of magnetic bead according to claim 1, it is characterized in that: the particle diameter of described surface-functionalized magnetic nano-particle is 5-250nm, described surface-functionalized magnetic nano-particle is the magnetic nano-particle of finishing or embeds the immobilized macromolecular magnetic nano-particle of macromolecular magnetic nano-particle or surface.
3. the preparation method of magnetic bead according to claim 1, is characterized in that: the surface functional group of described surface-functionalized magnetic nano-particle is one or more in hydroxyl or carboxyl or amino or aldehyde radical or sulfydryl.
4. the preparation method of magnetic bead according to claim 1, is characterized in that: described crosslinking agent adopts the macromolecular substances containing reactive group, and described reactive group is one or more in hydroxyl or carboxyl or aldehyde radical or sulfydryl or amino.
5. the preparation method of magnetic bead according to claim 4, is characterized in that: described macromolecular substances is one or more in polymer or block polymer or protein or DNA or polypeptide or polysaccharide.
6. the preparation method of magnetic bead according to claim 1, is characterized in that: the particle diameter of described magnetic bead is at least 2 times of described magnetic nano-particle particle diameter, is specially 200nm-50 μm.
7. the preparation method of magnetic bead according to claim 1, is characterized in that: described magnetic bead is spherical or irregular shape.
8. the preparation method of magnetic bead according to claim 1, is characterized in that: described magnetic bead complete disengaging time is in the solution 1min-6min.
9. the magnetic bead that obtains of the preparation method of the magnetic bead as described in any one of claim 1-8 is in biological medicine analysis and the application be separated, it is characterized in that: described magnetic bead is directly added in biological sample, utilize charge interaction to carry out the magnetic fast Acquisition separation and analysis of object;
Or first coupling capture agent on described magnetic bead, then the magnetic fast Acquisition separation and analysis carrying out object.
10. magnetic bead according to claim 9 is in biological medicine analysis and the application be separated, and it is characterized in that: described capture agent is antibody or fit.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410802284.6A CN104538168B (en) | 2014-12-23 | 2014-12-23 | A kind of preparation method and application of magnetic bead |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410802284.6A CN104538168B (en) | 2014-12-23 | 2014-12-23 | A kind of preparation method and application of magnetic bead |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104538168A true CN104538168A (en) | 2015-04-22 |
| CN104538168B CN104538168B (en) | 2019-05-03 |
Family
ID=52853678
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410802284.6A Active CN104538168B (en) | 2014-12-23 | 2014-12-23 | A kind of preparation method and application of magnetic bead |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104538168B (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104950026A (en) * | 2015-06-19 | 2015-09-30 | 苏州大学 | Electrochemical luminescence bioanalysis flow cell based on magnetic beads |
| CN107552021A (en) * | 2017-09-28 | 2018-01-09 | 青岛科技大学 | A kind of hydroxyl biomagnetic beads and its preparation method and application |
| CN109382052A (en) * | 2018-12-04 | 2019-02-26 | 郑州大学 | A kind of full-automatic coupling reaction device of visualization |
| CN110187104A (en) * | 2019-06-13 | 2019-08-30 | 华中农业大学 | Preparation method, sensor and its application of lateral relaxation time immunosensor based on bio-orthogonal reaction |
| CN111999158A (en) * | 2019-05-11 | 2020-11-27 | 南京岚煜生物科技有限公司 | Method for uniformly mixing magnetic beads |
| CN112213174A (en) * | 2019-07-12 | 2021-01-12 | 南京岚煜生物科技有限公司 | Method for uniformly mixing magnetic materials |
| US20210104343A1 (en) * | 2018-04-18 | 2021-04-08 | Sabic Global Technologies B.V. | Magnetic nanoparticles embedded in polymer microparticles |
| CN112979772A (en) * | 2021-03-28 | 2021-06-18 | 江苏集萃分子工程研究院有限公司 | Magnetic bead with low cleaning residue and preparation method thereof |
| CN112986349A (en) * | 2021-05-08 | 2021-06-18 | 江苏集萃分子工程研究院有限公司 | Magnetic bead modification labeling method for electrochemiluminescence detection |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1375507A (en) * | 2001-03-20 | 2002-10-23 | 清华大学 | Surface cladding and radical functino modification method of magnetic microsphere, thus obtained microsphere and its application |
| CN1931913A (en) * | 2005-09-14 | 2007-03-21 | 中国科学院过程工程研究所 | Surface functionalizing process of magnetic polystyrene microsphere |
| CN103495185A (en) * | 2013-09-02 | 2014-01-08 | 东华大学 | Preparation method of functionalized polyethyleneimine-modified multi-wall carbon nano-tube magnetic resonance imaging contrast agent |
-
2014
- 2014-12-23 CN CN201410802284.6A patent/CN104538168B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1375507A (en) * | 2001-03-20 | 2002-10-23 | 清华大学 | Surface cladding and radical functino modification method of magnetic microsphere, thus obtained microsphere and its application |
| CN1931913A (en) * | 2005-09-14 | 2007-03-21 | 中国科学院过程工程研究所 | Surface functionalizing process of magnetic polystyrene microsphere |
| CN103495185A (en) * | 2013-09-02 | 2014-01-08 | 东华大学 | Preparation method of functionalized polyethyleneimine-modified multi-wall carbon nano-tube magnetic resonance imaging contrast agent |
Non-Patent Citations (2)
| Title |
|---|
| 饶通德: "磁性纳米粒子的制备、表面功能化及应用研究", 《重庆大学硕士学位论文》 * |
| 黄天天等: "蛋白功能化磁性纳米颗粒的制备及应用", 《化学进展》 * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104950026B (en) * | 2015-06-19 | 2017-07-28 | 苏州大学 | Electrochemical luminescence bioanalysis flow cell based on magnetic bead |
| CN104950026A (en) * | 2015-06-19 | 2015-09-30 | 苏州大学 | Electrochemical luminescence bioanalysis flow cell based on magnetic beads |
| CN107552021B (en) * | 2017-09-28 | 2020-05-15 | 青岛科技大学 | Hydroxyl biomagnetic bead and preparation method and application thereof |
| CN107552021A (en) * | 2017-09-28 | 2018-01-09 | 青岛科技大学 | A kind of hydroxyl biomagnetic beads and its preparation method and application |
| US20210104343A1 (en) * | 2018-04-18 | 2021-04-08 | Sabic Global Technologies B.V. | Magnetic nanoparticles embedded in polymer microparticles |
| CN109382052A (en) * | 2018-12-04 | 2019-02-26 | 郑州大学 | A kind of full-automatic coupling reaction device of visualization |
| CN109382052B (en) * | 2018-12-04 | 2020-09-01 | 郑州大学 | Visual full-automatic coupling reaction device |
| CN111999158A (en) * | 2019-05-11 | 2020-11-27 | 南京岚煜生物科技有限公司 | Method for uniformly mixing magnetic beads |
| CN110187104A (en) * | 2019-06-13 | 2019-08-30 | 华中农业大学 | Preparation method, sensor and its application of lateral relaxation time immunosensor based on bio-orthogonal reaction |
| CN110187104B (en) * | 2019-06-13 | 2021-09-14 | 华中农业大学 | Preparation method of transverse relaxation time immunosensor based on bioorthogonal reaction, sensor and application thereof |
| CN112213174A (en) * | 2019-07-12 | 2021-01-12 | 南京岚煜生物科技有限公司 | Method for uniformly mixing magnetic materials |
| CN112979772A (en) * | 2021-03-28 | 2021-06-18 | 江苏集萃分子工程研究院有限公司 | Magnetic bead with low cleaning residue and preparation method thereof |
| CN112986349A (en) * | 2021-05-08 | 2021-06-18 | 江苏集萃分子工程研究院有限公司 | Magnetic bead modification labeling method for electrochemiluminescence detection |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104538168B (en) | 2019-05-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104538168A (en) | Magnetic bead preparing method and application | |
| CN103134926B (en) | Magnetic microsphere carrier and its making method | |
| CN1215902C (en) | Magnetic fluorescent double functional microballoon with core-shell structure and preparation method thereof | |
| KR101729687B1 (en) | Method for manufacturing suprerparamagnetic nanocomposite and suprerparamagnetic nanocomposite manufactured by the method | |
| JP6800275B2 (en) | Resin-platinum composite and its use | |
| CN1186377C (en) | Multifunctional organic-inorganic composite polymeric microball and preparing method thereof | |
| CN103275934A (en) | Separation method of micro circulating tumor cells | |
| CN102861541B (en) | Preparation method of surface-modified fluorescent magnetic polymer composite microspheres | |
| CN103305464B (en) | Method for directly separating CD<4+> and CD<8+> lymphocytes | |
| US20190170758A1 (en) | Method of detection with a fluorescent labeling particle | |
| Ding et al. | Aptamer-based nanostructured interfaces for the detection and release of circulating tumor cells | |
| CN102029133B (en) | Porous polymer microsphere and functional composite polymer microsphere as well as preparation method and application thereof | |
| CN103396786A (en) | Reversibly assembled and decomposable fluorescent magnetic nano material and nano biological probe and preparation methods thereof | |
| JP2010062444A (en) | Temperature-responsive magnetic fine particle capable of freeze drying | |
| JP2009168495A (en) | Colored latex | |
| CN105556289B (en) | The detection method of target substance | |
| Bone et al. | Physisorption and chemisorption of T4 bacteriophages on amino functionalized silica particles | |
| CN1947848A (en) | Functional magnetic separating rod and its making method | |
| Zhang et al. | Functionalization of high-moment magnetic nanodisks for cell manipulation and separation | |
| Fang et al. | A blood compatible, high-efficient sensor for detection of Cr (VI) in whole blood | |
| Reymond et al. | Fabrication and characterization of tosyl‐activated magnetic and nonmagnetic monodisperse microspheres for use in microfluic‐based ferritin immunoassay | |
| CN103305463B (en) | Method for separating CD4<+> and CD25<+> lymphocytes in human peripheral blood by use of magnetic beads | |
| CN103289954B (en) | Method for separating hematopoietic stem cells from human peripheral blood | |
| CN103657608B (en) | The preparation method of magnetic bag silicon yeast grafting polymine Biocomposite material and application thereof | |
| CN116053017B (en) | Composite magnetic microsphere and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |