CN104538168B - A kind of preparation method and application of magnetic bead - Google Patents
A kind of preparation method and application of magnetic bead Download PDFInfo
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- CN104538168B CN104538168B CN201410802284.6A CN201410802284A CN104538168B CN 104538168 B CN104538168 B CN 104538168B CN 201410802284 A CN201410802284 A CN 201410802284A CN 104538168 B CN104538168 B CN 104538168B
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Abstract
The invention discloses a kind of preparation method and applications of magnetic bead, belong to magnetic material preparation technical field.The preparation method is to prepare surface-functionalized magnetic nano-particle first, is then connected at least two surface-functionalized magnetic nano-particles using crosslinking agent, prepares the magnetic bead that magnetic content is 70%-98%.It is low and the problem of in order to realize quick separating, greatly increase magnetic bead volume and suspension stability is caused to reduce that the present invention solves existing magnetic bead magnetic content.In addition, for captured component biological respinse is especially immunoreacted, steric hindrance caused by small size magnetic bead produced by the present invention significantly reduces, capture ability and analysis detection sensitivity improve, it can be preferably applied to biological medicine separation and analysis, the rapid automatized detection of clinical biochemical especially suitable for the separation of cell, bacterium, virus, DNA/RNA, protein in biological fluid or reaction solution etc. and based on chemiluminescence, electrochemical luminescence and fluorescence.
Description
Technical field
The invention belongs to magnetic material preparation technical field more particularly to a kind of preparation method and applications of magnetic bead.
Background technique
For the magnetic nano-particle of conventional method synthesis, following both sides notable feature is typically exhibited.First, table
Reveal superparamagnetism or approximate superparamagnetism (remanent magnetism is less or few), is conducive to gather from the solution of dispersion under magnetic fields
It accumulates and separates, while being also beneficial to be dispersed back into solution after cancelling magnetic fields, and realize the analysis to separation component, on
It states feature and is particularly conducive to automation separation and analysis；Second, partial size (referring to the non-hydrated partial size characterized with TEM) is smaller, usually
In 10-100nm, each particle shows lesser magnetic force under strong magnetic field action, from the solution of dispersion disengaging time it is long,
Not exclusively, so as to cause analysis speed, slow, sensitivity for analysis is reduced for separation.
In order to using above-mentioned advantage (superparamagnetism or approximate superparamagnetism) and overcome disadvantages mentioned above (disengaging time be long, divides
From not exclusively), it is necessary to multiple magnetic nano-particles are assembled into the bigger magnetic bead of partial size (for the side of following expression
Just, magnetic nano-particle refers to that not connected, particle, magnetic bead refer to after connecting, large-sized
Particle).The magnetic force of one magnetic bead is to be summed it up by the magnetic force of multiple magnetic nano particles, makes the magnetic force of magnetic bead much larger than magnetic nano particle
Son, can quickly gather under magnetic fields in the solution of dispersion quickly, be kept completely separate.Simultaneously as magnetic bead be by
Magnetic nano-particle assembles, that is, remains the superparamagnetism or approximate superparamagnetism of magnetic nano-particle, cancelling magnetic field
The rapid dispersion again of magnetic bead can be realized after effect.For the magnetic bead that bio-separation and analysis use, there are also another in fact
Important requirement must have good suspension stability in the solution, otherwise cannot react with by separation component in certain
It carries out reacting quickly, completely in time and under suspended state.Therefore, for the bio-separation and analysis of a performance brilliance
Using magnetic bead must have three for the use of condition, first, strong magnetic force, preferably in common strong magnet (neodymium iron in 1-2 minutes
Boron magnets) under effect, realize dispersion magnetic bead in the solution it is quick, be kept completely separate；Second, superparamagnetism or approximate superparamagnetic
Property, so that magnetic bead (is such as shaken, liquid flowing) rapid dispersion again under weaker external force；Third, good suspension stability,
In aqueous solution can be suspending stabilized at least 30 minutes, it does not settle.
For existing magnetic bead, usually there are contradictions between strong magnetic force and suspension stability.In order to obtain strong magnetic force, it is necessary to
The quantity of magnetic nano-particle in magnetic bead is improved, but the volume of magnetic bead often quickly increases, and suspension stability is caused to reduce,
Magnetic bead is also caused to increase the steric hindrance of capture object simultaneously, capture ability reduces.Magnetic bead volume quickly increase it is main because
On the one hand element is the accumulation of magnetic nano particle, be on the other hand for the non-magnetic components of immobilized magnetic nano particle chemistry it is poly-
Product.The former cannot avoid, and therefore, solving unique method that magnetic bead quickly increases is to reduce the latter's containing in magnetic bead
Amount improves the magnetic content (content that magnetic content is magnetisable material in magnetic bead (magnetic nano particle)) of magnetic bead.Existing commodity
Change magnetic bead, including Medical magnetic ball, most of is high molecular polymer magnetic bead, including homopolymer and copolymer magnetic bead.Commonly
Synthesis macromolecule include polystyrene, polyacrylic acid, polymethylacrylic acid, polyethylene glycol, polyvinylpyrrolidone, polylactic acid,
Polyvinyl alcohol and their copolymer etc..Common preparation method includes investment, dispersion copolymerization method, emulsion polymerization and work
Property polymerization etc..These methods are to be coated on magnetic nano particle during polymerization reaction occurs for polymerizable organic monomer
Among polymer microsphere, to make immobilized multiple magnetic nano particles in a macromolecule magnetic bead.In order to allow magnetic nano particle to stablize
Ground is immobilized, must be requested that each of magnetic bead magnetic nano particle has been intended to enough polymeric PTC materials layers, results in this way
The increase of namagnetic substance content in magnetic bead, i.e. magnetic content are lower.Other than macromolecule magnetic bead, in fact, going back organic and/or inorganic materials
Magnetic bead.SiO is prepared using the method for hydrolysis organosiloxane2The magnetic bead of cladding, preparation method mainly include reverse microemulsion method, gas
Colloidal sol high-temperature decomposition and sol-gal process etc..Under the premise of guaranteeing magnetic force, the magnetic content of this kind of magnetic bead is lower.Above-mentioned two class
Magnetic bead is during preparing non-magnetic microspheres by cladding (including physics cladding and the cladding immobilized based on chemical bond)
Immobilized magnetic nano particle, this is the content for inevitably increasing namagnetic substance, so that reduction prepares the magnetic content of magnetic bead, this
Be in one intrinsic contradictions, it is difficult to avoided from method.
Summary of the invention
The purpose of the present invention is to propose to a kind of preparation method of magnetic bead, the preparation method using crosslinking agent reactive group and
Chemical reaction between the surface functional group of magnetic nano-particle is formed by chemical bond, and multiple magnetic nano-particles are connected
Get up, prepares the magnetic bead of high magnetic content.
To achieve this purpose, the present invention adopts the following technical scheme:
Invention provides a kind of preparation method of magnetic bead, prepares surface-functionalized magnetic nano-particle first, then adopts
At least two surface-functionalized magnetic nano-particles are connected with crosslinking agent, preparing magnetic content is 70%-98%'s
Magnetic bead.
As a preferred embodiment, the partial size of the surface-functionalized magnetic nano-particle is 5-250nm, the surface
The magnetic nano-particle of functionalization is the magnetic nano-particle of surface modification or magnetic nano-particle or the surface for embedding macromolecular
The magnetic nano-particle of immobilized macromolecular.
As a preferred embodiment, the surface functional group of the surface-functionalized magnetic nano-particle is hydroxyl or carboxylic
Base or amino or one or more of aldehyde radical or sulfydryl.
As a preferred embodiment, the crosslinking agent uses the macromolecular substances containing reactive group, the reactive group
For hydroxy or carboxy or aldehyde radical or one or more of sulfydryl or amino.
As a preferred embodiment, the macromolecular substances are polymer or block polymer or protein or DNA or more
One or more of peptide or polysaccharide.
As a preferred embodiment, the partial size of the magnetic bead is at least 2 times of the magnetic nano-particle partial size, specially
200nm-50μm。
As a preferred embodiment, the magnetic bead is spherical or irregular shape.
As a preferred embodiment, the time that is kept completely separate of the magnetic bead in the solution is 1min-6min.
The present invention also provides magnetic beads made from the preparation method of a kind of as above any magnetic bead in biological medicine point
Analyse with isolated application, the magnetic bead is directly added into biological sample, utilize charge interaction carry out object magnetism
Fast Acquisition separation and analysis；
Or the coupling capture reagent first on the magnetic bead, then carry out the magnetic fast Acquisition separation of object and divide
Analysis.
As a preferred embodiment, the capture reagent is antibody or aptamer.
The invention has the benefit that
The present invention provides a kind of preparation method of magnetic bead, the present invention utilizes the reactive group and magnetic nano particle of crosslinking agent
Chemical reaction between the surface functional group of son is formed by chemical bond, and multiple magnetic nano-particles are connected, and prepares
The magnetic bead of high magnetic content (70%-98%) out.Preparation method of the present invention is simple, prepares yield height, and magnetic bead particle size is controllable；?
Prepare immobilized same amount of magnetic nano particle in magnetic bead, i.e., it is prepared by the present invention under the premise of each magnetic bead has identical magnetic force
The magnetic content of magnetic bead is higher, and volume is smaller, and surface charge is abundant, and surface hydrophilicity is stronger, and suspension stability is more preferable, solves existing
There is the magnetic bead especially low problem of Medical magnetic ball magnetic content, also overcomes existing magnetic bead to realize quick separating, greatly increase
The problem of adding magnetic bead volume and suspension stability caused to reduce.
The present invention also provides magnetic beads made from a kind of preparation method of above-mentioned magnetic bead to analyze and separate in biological medicine
Application, the magnetic bead is directly added into biological sample, utilize charge interaction carry out object magnetic fast Acquisition
Separation and analysis；Or first on the magnetic bead coupling capture reagent, then carry out object magnetic fast Acquisition separation with
Analysis.For captured component biological respinse is especially immunoreacted, caused by small size magnetic bead produced by the present invention
Steric hindrance significantly reduces, and capture ability and analysis detection sensitivity improve, and can be preferably applied to biological medicine separation and divide
Analysis, especially suitable for cell, bacterium, virus, DNA/RNA, protein in biological fluid or reaction solution etc. separation and be based on
The rapid automatized detection of the clinical biochemical of chemiluminescence, electrochemical luminescence and fluorescence.
Detailed description of the invention
Fig. 1 is the thermogravimetric curve figure of the PEI crosslinking magnetic bead of the embodiment of the present invention one；
Fig. 2 is the PEI crosslinking magnetic bead of the embodiment of the present invention one compared with certain popular Medical magnetic ball, the magnetic bead in Magneto separate
Surplus ratio (not by the percentage of separation magnetic bead in solution) versus time curve figure；
Fig. 3 is that the mould of the ultra-violet absorption spectrum of the PEI crosslinking magnetic bead solution of the embodiment of the present invention one changes with time song
Line chart.
Specific embodiment
With reference to embodiment, the embodiment of the present invention is furthur described in detail.Following embodiment is used for
Illustrate the present invention, but is not intended to limit the scope of the invention.
To keep the technical problems solved, the adopted technical scheme and the technical effect achieved by the invention clearer, below
It will the technical scheme of the embodiment of the invention will be described in further detail in conjunction with attached drawing, it is clear that described embodiment is only
It is a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, those skilled in the art exist
Every other embodiment obtained under the premise of creative work is not made, shall fall within the protection scope of the present invention.
It is well known that surface-functionalized magnetic nano-particle be broadly divided into it is following several:
1, the magnetic nano-particle of non-surface modification: the magnetic nano-particle prepared with conventional method, including Fe3O4、
Fe2O3And its their heterozygote, CoFe2O4、NiFeO4、ZnFe2O4、MnFeO4Deng numerous functional groups on surface refer to largely
Hydroxyl (- OH).
2, it the magnetic nano-particle of surface modification: for the magnetic nano-particle prepared with conventional method, is repaired by chemistry
Decorations change the functional group of particle surface, and corresponding functional group can be its of hydroxy or carboxy or amino or aldehyde radical or sulfydryl
Middle one or more.
3, it embeds the magnetic nano-particle of macromolecular: while preparing magnetic nano-particle with conventional method, being added and need
The macromolecular of embedding makes its macromolecule fraction be embedded in inside particles, and another part is outside particle.But it is big to be not excluded for some
Molecule is to be adsorbed on the surface of the particle.The reactive group of macromolecular outside particle is surface functional group.
4, the magnetic nano-particle of the immobilized macromolecular in surface: after preparing magnetic nano-particle with conventional method, change is utilized
Learn one or more of key, hydrophobic effect, charge effect active force macromolecular is immobilized in particle surface.But it is not excluded for some
Macromolecular is to be adsorbed on the surface of the particle.The reactive group of macromolecular outside particle is surface functional group.
Embodiment one
Present embodiments provide a kind of preparation method of magnetic bead, comprising the following steps:
(1) the magnetic nano-particle preparation of citric acid modification: by the Fe of preparation3O4Magnetic nano-particle aqueous-based dispersions are super
Sound disperses 15min, takes 20mL to be placed in three-necked bottle and leads to nitrogen deoxygenation 20min, is added in a certain amount of sodium citrate solution and surpasses
Sound 15min, makes Fe3O4Magnetic nano-particle is uniformly dispersed in system, connects experimental provision, keeps nitrogen environment, machinery is stirred
The speed mixed is adjusted to 600rpb, and water bath temperature is stirred to react 1h after rising to 60 DEG C, carries out Magneto separate with ndfeb magnet, three steam
Water washing removes extra sodium citrate, until Magneto separate cannot obtain clear supernatant, is finally scattered in again
In 20mL tri-distilled water, the water-soluble Fe of citric acid modification is made3O4, i.e. the magnetic nano-particle of citric acid modification, surface functional group
Group is carboxyl.
(2) the PEI crosslinking of magnetic nano-particle prepares magnetic bead: in the aqueous-based dispersions of 15mg magnetic nano-particle, adopting
With EDCHCl and sulfo-NHS activated carboxyl, shaking table activates 1h under 37 DEG C of reaction condition, Magneto separate, washing three times, by it
The ultrasonic disperse 1min under 37 DEG C of water bath conditions is added in round-bottomed flask；Take the polymer P EI (Mw=1800) of 12.5mg water-soluble
In a round bottom flask, and the buffer solution that pH=7.4 is added is placed in shaking table and shakes liquid, reacts 1h, and Magneto separate is washed, multiple
Magnetic bead soluble in water to get PEI crosslinking.
The partial size and Zeta potential of magnetic bead manufactured in the present embodiment are analyzed:
It can be seen that the partial size for dynamic light scattering determination from subordinate list 1, magnetic bead manufactured in the present embodiment
ChiSquared value and P.I. value are smaller, illustrate that particle diameter distribution is satisfied with normal distribution well, and be evenly distributed.Also,
Magnetic bead has surface charge abundant, and corresponding partial size is much smaller than the partial size (about 2700nm) of existing Medical magnetic ball.Above-mentioned spy
Sign makes it show good suspension stability in aqueous solution.More importantly small scale magnetic bead will greatly reduce
Magnetic bead increases capture ability to the steric hindrance of captured component, improves detection sensitivity, reduces capture time, realizes quick
Analysis detection.
The partial size and Zeta potential of the magnetic bead of the PEI preparation of table 1Mw=1800
The thermogravimetric curve and magnetic content analysis of magnetic bead manufactured in the present embodiment:
We have done thermal weight loss characterization to the magnetic bead of PEI crosslinking (Mw=1800) preparation, as shown in Fig. 1, from thermogravimetric song
It will be seen that weightlessness is in two stages on line: the first stage is temperature at 140 DEG C or less, and the weightlessness of this section is
Water evaporation in magnetic bead surface layer, magnetic bead magnetic content is 99.12% at this time；Second stage is since 200 DEG C, this is because magnetic
Weightlessness caused by non-magnetic components contained in pearl (including surface-functionalized citric acid and macromolecular PEI) decompose.Temperature exists
After 640 DEG C, thermogravimetric curve tends towards stability, and is remaining Fe3O4.With the thermal weight loss of the non-magnetic components including moisture content
Based on rate, the magnetic content for calculating magnetic bead is 91.87%.Certainly, the influence of moisture content is removed, magnetic content can be higher, this extremely has
It is prepared conducive to the efficient magnetic bead of strong magnetic force, small size, higher suspension stability.
Magnetic bead manufactured in the present embodiment is compared with the Magneto separate of Medical magnetic ball:
In Magneto separate, with by the percentage of the total magnetic bead of separation magnetic bead Zhan, (present invention is not defined as " magnetic bead residue in solution
Rate ") it is standard, it investigates with the Magneto separate ability of PEI crosslinking magnetic bead and certain popular Medical magnetic ball.From attached drawing 2 as it can be seen that adding magnetic outside
When field action 1min, not by isolated surplus ratio, Medical magnetic ball is slightly below the PEI crosslinking magnetic bead of the present embodiment, but after 2min
The surplus ratio of PEI crosslinking magnetic bead is just less than the surplus ratio of Medical magnetic ball.PEI crosslinking magnetic bead realizes intimate 100% in 6min
Separation, still, Medical magnetic ball can not achieve 100% separation for a long time, and surplus is up to 10% or more, this has just been implied
Up to 10% or more can not achieve separation by separation component.For the highly sensitive luminesceence analysis in clinical examination (as extensively
The chemiluminescence that uses, electrochemiluminescence analysis) for, imply that false negative will occur in a large amount of low content sample.This for
The biomarker analyte detection of now clinical widely used early diagnosis of cancer, be it is extremely disadvantageous, easily there are a large amount of missing inspections.
It is indicated above that magnetic bead manufactured in the present embodiment, Magneto separate is much better than certain popular Medical magnetic ball.
The suspension stability of magnetic bead and Medical magnetic ball manufactured in the present embodiment compares:
Since the ultraviolet spectra of magnetic bead does not have characteristic absorption, table is come with the mould versus time curve of ultraviolet light absorption spectrum
Levy the magnetic bead content in solution.Still using certain popular Medical magnetic ball as standard of comparison, from attached drawing 3 as it can be seen that in 20min two class magnetic beads
Significant difference is not present in suspension stability in the solution, shows good stability.But with the increasing of time
Add, the difference of the two is just extremely obvious, when being up to 120nm, magnetic bead manufactured in the present embodiment be still it is extremely stable, do not have
It was found that apparent sedimentation, but the Medical magnetic ball compareed has settled 15%.It is fabulous to show that magnetic bead manufactured in the present embodiment has
Suspension stability is much better than certain popular Medical magnetic ball.
Embodiment two:
The basic phase of preparation method of a kind of preparation method of magnetic bead provided in this embodiment and magnetic bead described in embodiment one
Together, difference place is:
(1) functional group (packet the preparation of the magnetic nano-particle of the surface modification with small molecule functional group: will be contained
Include hydroxyl, amino, aldehyde radical, sulfydryl and the carboxyl contained by different from other carboxylic acids of citric acid) Small Molecule Functionalization reagent,
Instead of the citric acid of step (1) in embodiment one, the surface modification of similar magnetic nano-particle is carried out to get with different function
The magnetic nano-particle for the surface modification that can be rolled into a ball.
(2) preparation of the magnetic nano-particle of the immobilized macromolecular in surface: the macromolecular that surface will be needed immobilized, including polymerization
Object macromolecular is replaced in (1) of the present embodiment with block polymer, protein, DNA, polypeptide, polysaccharide of chemistry key connection etc.
Small Molecule Functionalization reagent, with similar method to get the magnetic nano-particle of the immobilized macromolecular in surface.
(3) preparation of the magnetic bead of macromolecules cross-linking: by macromolecular include polymer macromolecule, with chemistry key connection block
Polymer, protein, DNA, polypeptide, polysaccharide etc. replace the polymer P EI of step (2) in embodiment one, according to similar method
The magnetic bead of different macromolecules cross-linkings can be prepared.
Embodiment three
The basic phase of preparation method of a kind of preparation method of magnetic bead provided in this embodiment and magnetic bead described in embodiment one
Together, difference place is:
(1) it embeds the magnetic nano-particle preparation of PEI: taking the tri-distilled water of 80mL in the four-necked bottle of 250mL, logical nitrogen removes
After oxygen 20min, experimental provision is connected, checks air-tightness, guarantees that air-tightness is good；Bath temperature is adjusted to 35 DEG C, machinery stirs
The revolving speed mixed is adjusted to 300rpb, keeps the nitrogen environment in four-necked bottle, accurately weighs the FeCl of 4.05g3·6H2O's and 1.98g
FeCl2·4H2O is added to stirring and dissolving 20min in four-necked bottle, and a certain amount of polymerization is added in certain density ammonia spirit
Churned mechanically revolving speed is adjusted to 600rpb by object macromolecular PEI (Mw=10000), and ammonium hydroxide dilution is slowly added dropwise until solution
PH=11 stops that ammonium hydroxide is added dropwise；The reaction was continued after bath temperature is risen to 60 DEG C 1h.
After reaction, the suspension of black is transferred in beaker, Magneto separate, abandons supernatant, washed with tri-distilled water redissolution
It washs, Magneto separate, abandons supernatant liquor, so repeat, until solution is in neutrality；The Fe that will be obtained3O4Nanoparticle is dispersed in 100mL's
Ultrasound 30min in tri-distilled water stands up to the magnetic nano-particle of PEI is embedded.
(2) the PEI crosslinking of magnetic nano-particle prepares magnetic bead: using EDCHCl and sulfo-NHS activated polyacrylic acid
In carboxyl, under conditions of 37 DEG C shaking table activate 1h, be added into it is a certain amount of embedding PEI magnetic nano-particle moisture
Ultrasonic disperse 1min in dispersion liquid is placed in shaking reaction 1h in shaking table under 37 DEG C of water bath conditions, and Magneto separate, washing is multiple soluble in water,
Up to magnetic bead.
In addition, can will be containing a large amount of reactive groups (including hydroxyl, carboxylic acid, aldehyde radical, sulfydryl and different from the other poly- of PEI
Close organic amine contained by amino) macromolecular replace the present embodiment (1) in PEI, carry out similar macromolecular embedding, i.e.,
There must be the magnetic nano-particle of different embedding macromoleculars.
Example IV
Present embodiments provide a kind of preparation method of magnetic bead as described in embodiment one or embodiment two or embodiment three
Magnetic bead is directly added into biological sample in biological medicine analysis and isolated application, utilizes charge phase interaction by magnetic bead obtained
It is separated and is analyzed with the magnetic fast Acquisition for carrying out object；Or the coupling capture reagent first on magnetic bead, then carry out target
The separation of magnetic fast Acquisition and analysis of object.
Now illustrate by taking magnetic bead of the surface of embodiment three with carboxyl as an example.By the magnetism EDC-NHS of 50 μ L 5mg/mL
Method activated carboxyl, Magneto separate redissolve in the EP pipe of 1.5mL albumen low adsorption, add the PBS of 200ul0.01M pH7.4, whirlpool
Rotation is allowed to uniformly, 50 μ L, 40 μ g/mL rabbit anti-mouse IgG be added and hatches 30min, and 1% BSA that 50 μ L are added closes 30min,
Magneto separate, the PBS that 300 μ L 0.01M pH7.4 are added redissolve to get the immune magnetic of coupling capture reagent rabbit anti-mouse IgG antibody
Pearl.
For captured component biological respinse is especially immunoreacted, caused by small size magnetic bead produced by the present invention
Steric hindrance significantly reduce, capture ability and analysis detection sensitivity improve, can be preferably applied to biological medicine separation with
Analysis, separation and base especially suitable for cell, bacterium, virus, DNA/RNA, protein in biological fluid or reaction solution etc.
In the rapid automatized detection of the clinical biochemical of chemiluminescence, electrochemical luminescence and fluorescence.
The particle size range of the surface-functionalized magnetic nano-particle of the present invention is 5-250nm, the surface-functionalized magnetic
Property nanoparticle can for surface modification magnetic nano-particle or embed macromolecular magnetic nano-particle or surface it is immobilized big
The magnetic nano-particle of molecule.The partial size of magnetic bead is at least 2 times of magnetic nano-particle partial size, and specific range is 200nm-50 μ
M, i.e. the hydration partial size of the magnetic bead of dynamic light scattering determination are 200nm or more, maximum up to 50 μm, and the form of magnetic bead is ball
Shape or irregular shape.The time that is kept completely separate of magnetic bead in the solution controls in 1min-6min range, the i.e. quick separating of magnetic bead
Between most it is short be 1min.
The present invention is using between the reactive group of crosslinking agent and the surface functional group of at least two magnetic nano-particles
Chemical reaction is formed by chemical bond, and multiple magnetic nano-particles are connected, high magnetic content (70%-98%) is prepared
Magnetic bead.
Preparation method of the present invention is simple, prepares yield height, and magnetic bead particle size is controllable；The immobilized equal amount in preparing magnetic bead
Magnetic nano particle, i.e., each magnetic bead have identical magnetic force under the premise of, the magnetic content of magnetic bead prepared by the present invention is higher, volume
Smaller, surface charge is abundant, and surface hydrophilicity is stronger, and suspension stability is more preferable, solves existing magnetic bead especially Medical magnetic ball
The low problem of magnetic content also overcomes existing magnetic bead to realize quick separating, greatly increases magnetic bead volume and cause to suspend
The problem of stability reduces.
The technical principle of the invention is described above in combination with a specific embodiment.These descriptions are intended merely to explain of the invention
Principle, and shall not be construed in any way as a limitation of the scope of protection of the invention.Based on the explanation herein, the technology of this field
Personnel can associate with other specific embodiments of the invention without creative labor, these modes are fallen within
Within protection scope of the present invention.
Claims (6)
1. a kind of preparation method of magnetic bead, it is characterised in that: prepare surface-functionalized magnetic nano-particle first, then use
Crosslinking agent connects at least two surface-functionalized magnetic nano-particles, prepares the magnetic that magnetic content is 70%-98%
Pearl；The surface-functionalized magnetic nano-particle is the magnetic nano-particle of citric acid modification；The crosslinking agent is Mw=
1800 polyetherimide.
2. the preparation method of magnetic bead according to claim 1, it is characterised in that: the partial size of the magnetic bead is at least the magnetic
2 times, specially 200nm-50 μm of property nano particle diameter.
3. the preparation method of magnetic bead according to claim 1, it is characterised in that: the magnetic bead is spherical or irregular shape.
4. the preparation method of magnetic bead according to claim 1, it is characterised in that: the magnetic bead being kept completely separate in the solution
Time is 1min-6min.
5. magnetic bead made from a kind of preparation method of magnetic bead according to any one of claims 1-4 is analyzed and is divided in biological medicine
From application, it is characterised in that: the magnetic bead is directly added into biological sample, charge interaction is utilized to carry out object
Magnetic fast Acquisition separation and analysis；
Or the coupling capture reagent first on the magnetic bead, then carry out the separation of magnetic fast Acquisition and analysis of object.
6. magnetic bead according to claim 5 is analyzed and isolated application in biological medicine, it is characterised in that: the capture examination
Agent is antibody or aptamer.
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| CN107552021B (en) * | 2017-09-28 | 2020-05-15 | 青岛科技大学 | Hydroxyl biomagnetic bead and preparation method and application thereof |
| CN112004864A (en) * | 2018-04-18 | 2020-11-27 | 沙特基础工业全球技术公司 | Magnetic nanoparticles embedded in polymer microparticles |
| 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 |
| 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 |
| CN112979772B (en) * | 2021-03-28 | 2022-06-24 | 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 |
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