CN106237947B - The magnetic microsphere and preparation method thereof of high density carboxyl modified - Google Patents
The magnetic microsphere and preparation method thereof of high density carboxyl modified Download PDFInfo
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- CN106237947B CN106237947B CN201610780210.6A CN201610780210A CN106237947B CN 106237947 B CN106237947 B CN 106237947B CN 201610780210 A CN201610780210 A CN 201610780210A CN 106237947 B CN106237947 B CN 106237947B
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
- B01J13/02—Making microcapsules or microballoons
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
- C12N15/1013—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers by using magnetic beads
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01F—MAGNETS; INDUCTANCES; TRANSFORMERS; SELECTION OF MATERIALS FOR THEIR MAGNETIC PROPERTIES
- H01F1/00—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties
- H01F1/42—Magnets or magnetic bodies characterised by the magnetic materials therefor; Selection of materials for their magnetic properties of organic or organo-metallic materials, e.g. graphene
Abstract
The present invention provides a kind of magnetic microspheres and preparation method thereof of high density carboxyl modified, and the preparation method comprises the following steps: preparing amido modified magnetic microsphere；The amido modified magnetic microsphere is mixed with organic carboxyl acid, adds EDC and NHS, reaction obtained the magnetic microsphere of the high density carboxyl modified after 1~3 hour.The present invention provides magnetic microsphere surface carboxyl have the characteristics that it is highdensity, the carboxyl on carboxyl modified magnetic microsphere surface of the invention simultaneously is coupled by amino and magnetic microsphere, its space arm lengths is longer than the space arm of the magnetic microsphere of general carboxyl modified, therefore the sequencable carboxyl density in magnetic microsphere surface is higher.Raw material of the present invention is simple and easy to get, at low cost, and processing step is simple, has very strong practicability.
Description
Technical field
The present invention relates to the preparation field of magnetic microsphere, the magnetic microsphere of specifically a kind of high density carboxyl modified and
Preparation method.
Background technique
Biomagnetism microballoon is a kind of novel functionalization solid carrier, the active group of surface modification, can with it is more
Kind of bioactive substance is coupled, with liquid mobility and solid magnetic material the features such as, in the effect of external magnetic field
Under can with displacement and concentration, after removing external magnetic field, slightly vibrate or aspirate again can be dispersed in liquid, to make
The separation of solid liquid phase becomes very efficient and convenient, by simply eluting the very high targeting substance of available purity.Using receiving
Micron superparamagnetism magnetic microsphere extracts the method for nucleic acid than traditional method (Chelex100 method, organic method, silica
Method, salting out method etc.) it is more simple, conveniently, and have the number of transfer centrifuge tube it is less, it is not easy to pollute, without centrifugation, operation
It is easy to be quick, the advantages that can be realized high throughput, automate.This method relies primarily on electrostatic interaction, hydrophobic effect and hydrogen bond and makees
It is integrated on magnetic microsphere with by nucleic acid.First it is released DNA/RNA therein by lysate effect cell or tissue
Come, then the magnetic microsphere Jing Guo surface modification is specifically bound with nucleic acid, forms nucleic acid-magnetic microsphere compound, most
Afterwards under the action of externally-applied magnetic field, nucleic acid-magnetic microsphere compound is isolated.
Wherein, amino and carboxyl are two kinds of active groups for being commonly used to modified magnetic microballoon, and under certain condition, they can
To obtain immunomagnetic beads with biological ligand cou.Wherein, amido modified magnetic microsphere is easy to get, and amino activity compared with
It is good, it is easy to carry out chemical coupling with biological aglucon.Penta 2 generally are utilized by glutaraldehyde and biological ligand cou when in use
Aldehyde carries out reductive amination process to the amino in the amino on magnetic microsphere surface and biological aglucon as space arm, obtains higher
Package amount.But this method also needs to add when carrying out reductive amination process strong reductant (such as sodium borohydride or cyano boron
Sodium hydride), albumen isoreactivity substance inactivation is easily caused, and glutaraldehyde also results in magnetic microsphere and biological aglucon itself hair
Raw crosslinking also results in protein interior crosslinking, is unfavorable for the coating of biological aglucon.
And the magnetic microsphere of carboxyl modified then utilizes the carboxyl and EDC (1- (3- dimethylamino-propyl)-on magnetic microsphere surface
3- ethyl carbodiimide)/NHS (n-hydroxysuccinimide) reaction after, generate NHS activation magnetic microsphere intermediate, then benefit
With the amino coupled on NHS group and biological aglucon.This method is easily operated, and is swift in response, and is suitable for small molecule, small peptide is made
For the coupling of biological aglucon.But the carboxyl density on the magnetic microsphere surface of carboxyl modified is low, active site is less, and carboxyl
Activity is low, coupling efficiency is low, the package amount of biological aglucon is few.
Summary of the invention
The main object of the present invention is to provide a kind of magnetism of high density carboxyl modified in view of the shortcomings of the prior art
The preparation method of microballoon, the preparation method is simple, and the carboxyl density on the magnetic microsphere surface being prepared is high, and active site is more.
In one aspect of the invention, the present invention is achieved through the following technical solutions: a kind of high density carboxyl modified
The preparation method of magnetic microsphere, it is characterised in that: the preparation method comprises the following steps:
S1: amido modified magnetic microsphere is prepared；
S2: the amido modified magnetic microsphere is mixed with organic carboxyl acid, adds EDC and NHS, is reacted 1~3 hour
The magnetic microsphere of the high density carboxyl modified is obtained afterwards.
The amido modified magnetic microsphere can be using commercial goods or according to known to those skilled in the art normal
The preparation of rule method, does not influence result of the invention.The present invention prepares the magnetic microsphere of amido modified Silica-coated, can
So that magnetic nano-particle is gathered in silica interior, core-shell structure is formed, increases partial size, and increases magnetic and stablizes
Property.Effect is more better than directly carrying out amido modified obtaining amido modified magnetic microsphere on magnetic nanoparticle surface.
Preferably, in the step s1, amido modified magnetic microsphere is the magnetism of amido modified Silica-coated
Microballoon, it is prepared via a method which to obtain: ammonium hydroxide, silylating reagent being added in the solution of Xiang Hanyou magnetic nano-particle
And amino silicane coupling agent, reaction obtained the magnetic microsphere of amido modified Silica-coated after 1~3 day, the magnetism is received
The mass ratio that rice corpuscles, ammonium hydroxide, silylating reagent, amino silicane coupling agent are added are as follows: 1:(12.5~40): (2~8): (0.5
~3)；The mass concentration of ammonium hydroxide is 25-28%.Wherein, silylating reagent can be tetraethyl orthosilicate (CAS:562-90-3),
Amino silicane coupling agent can be (3- aminopropyl) triethoxysilane (CAS:919-30-2).Those skilled in the art are selecting
When selecting other silylating reagents and amino silicane coupling agent, the ratio of reaction raw materials can be adjusted, without departing from this
The protection scope of invention.
Obtained amido modified magnetic microsphere can carry out conventional washing purifying, such as: by Magneto separate, and
Alternately washing is purified for water, ethyl alcohol.
Preferably, the magnetic nano-particle is magnetic Nano Fe3O4, it is to be made by the steps to obtain:
S1-1: in air, ammonium hydroxide is added into the aqueous solution of divalent iron salt, stirring keeps part ferrous oxidising for trivalent
Iron, oxidation precipitation generates black Fe under alkaline condition3O4Particle；
S1-2: oleic acid being added into step s1-1, after mixing heating reaction 3~5 hours at 60~130 DEG C, i.e.,
The super-paramagnetism nano Fe of the oleic acid package can be obtained3O4。
Magnetic nano-particle of the invention can also be prepared by other conventional methods, such as hydro-thermal method, solvent heat
Method, coprecipitation etc..
Preferably, in the step (2), the organic carboxyl acid be at least containing there are three or three or more carboxyls polynary carboxylic
Acid.After polybasic carboxylic acid is by one of carboxyl and the amido modified magnetic microsphere coupling, it there remains multiple carboxyls, it can
Increase the superficial density of carboxyl.
Most preferably, in the step (2), the polybasic carboxylic acid is polyacrylic acid, hyaluronic acid, polyglutamic acid or amber
Amic acid dendritic.
In order to guarantee organic carboxyl acid carboxyl can with the amino coupled on magnetic microsphere surface, while make organic carboxyl acid due to
The limitation of steric hindrance only one carboxyl and amino coupled as far as possible should control amido modified magnetic microsphere and organic carboxylic
The ratio of acid.
Preferably, in the step (2), the mass ratio of the amido modified magnetic microsphere and the organic carboxyl acid is 1:
(0.4~2).
It is furthermore preferred that in the step (2), the matter of EDC, NHS of addition, organic carboxyl acid, amido modified magnetic microsphere
Amount is than being (10~60): (5~30): (4~20): 10.
In the second aspect of the present invention, a kind of high density carboxyl modified being prepared according to any of the above-described method is provided
Magnetic microsphere.
In the third aspect of the present invention, a kind of nucleic acid extraction kit is provided, including magnetic microsphere, the magnetism
Microballoon is the magnetic microsphere of above-mentioned high density carboxyl modified.
In the fourth aspect of the present invention, a kind of immunomagnetic beads are provided, the immunomagnetic beads are by above-specified high density carboxylic
The magnetic microsphere of base modification and biological ligand cou, are prepared.
The biology aglucon can be the biologically active aglucon such as antibody, enzyme, BSA, DNA or RNA.
The invention has the benefit that
It (1) is by by amido modified magnetic microsphere surface the present invention provides a kind of magnetic microsphere of carboxyl modified
Amino and organic carboxyl acid be coupled, the carboxyl on magnetic microsphere surface has the characteristics that highdensity.
(2) since the amino expression activitiy on magnetic microsphere is high, the coupling with organic carboxyl acid is easy, and avoids use
Glutaraldehyde carries out the deficiency (there are internal crosslinking, reducing agent easily leads to albumen isoreactivity substance inactivation) of reductive amination process, makes
Reaction product is more controllable.
(3) carboxyl on carboxyl modified magnetic microsphere surface of the invention is coupled by amino and magnetic microsphere, empty
Between arm lengths it is longer than the space arm of the magnetic microsphere of general carboxyl modified, therefore the sequencable carboxyl in magnetic microsphere surface is close
Du Genggao, when extracting nucleic acid, since binding site is more, extraction efficiency is also higher.It can also will be of the invention highly dense
The magnetic microsphere and biological ligand cou for spending carboxyl modified, obtain immunomagnetic beads, since carboxyl density is high, the coating of biological aglucon
Amount is also bigger.
(4) raw material of the present invention is simple and easy to get, at low cost, and processing step is simple, has very strong practicability.
Detailed description of the invention
Fig. 1 is the SEM figure of the magnetic microsphere 1 of amido modified Silica-coated；
Fig. 2 is the TEM figure of the magnetic microsphere 1 of amido modified Silica-coated；
Fig. 3 is the electrophoretogram after the extraction of plasmid DNA of embodiment 4.
Specific embodiment
By the following specific examples further illustrate the invention: the experiment of actual conditions being not specified in the following example
Method according to conventional methods and conditions, or is selected according to product manual.
Ammoniacal liquor mass concentration used in the embodiment of the present invention is 25~28%.
Polyacrylic acid molecular weight used in the embodiment of the present invention is 2000, CAS:9003-01-4.
Hyaluronan molecule amount used in the embodiment of the present invention is 4000.
Polyglutamic acid molecular weight used in the embodiment of the present invention is 50000, CAS:25513-46-6.
Succinamic acid dendritic polymer molecule amount used in the embodiment of the present invention is 4885.40~83805.01, purchase
From ALDRICH.
Embodiment 1
(1) in air, by 7g FeCl2·4H2O is added in 50mL deionized water, and obtaining concentration is 0.14g/mL's
FeCl2Aqueous solution.To 50mL FeCl2Ammonium hydroxide 30mL is added in aqueous solution, after stirring 45min, color gradually becomes light green color, then
Green is deepened, black is eventually become；
(2) 1.1g oleic acid is added into step (1), after mixing sets mixed liquor in closed reaction kettle, 110
Heating reaction 4 hours at DEG C, are then alternately washed with deionized water and ethyl alcohol each primary, are dispersed in n-hexane after Magneto separate,
The paramagnetic nanoparticles Fe of the oleic acid package of black can be obtained3O41；
(3) the paramagnetic nanoparticles Fe of Xiang Hanyou 10mg oleic acid package3O4125mg ammonium hydroxide, the positive silicon of 30mg are added in 1 solution
Sour tetra-ethyl ester and 30mg (3- aminopropyl) triethoxysilane, reaction carries out Magneto separate after 1 day, and is replaced with second alcohol and water
Washing respectively obtains the magnetic microsphere 1 of amido modified Silica-coated afterwards twice.
(4) by the magnetic microsphere 1 of 10mg step (3) the amido modified Silica-coated and 10mg polypropylene acid-mixed
It closes, adds 60mg EDC and 30mg NHS, reaction obtained the magnetic microsphere 1 of the high density carboxyl modified after 1 hour.
Embodiment 2
(1) in air, by 10g FeCl2·H2O is added in 100mL deionized water, and obtaining concentration is 0.10g/mL's
FeCl2Aqueous solution.To 100mLFeCl2·H2Ammonium hydroxide 20mL is added in O aqueous solution, after stirring 60min, color gradually becomes light green
Color, then green is deepened, eventually become black；
(2) 1g oleic acid is added into step (1), after mixing sets mixed liquor in closed reaction kettle, at 60 DEG C
Lower heating is reacted 5 hours, and then with deionized water and ethyl alcohol, alternately washing is respectively dispersed in pungent hexane twice, after Magneto separate, i.e.,
The paramagnetic nanoparticles Fe of the oleic acid package of black can be obtained3O42；
(3) the paramagnetic nanoparticles Fe of Xiang Hanyou 10mg oleic acid package3O4250mg ammonium hydroxide, the positive silicon of 20mg are added in 2 solution
Sour tetra-ethyl ester and 10mg (3- aminopropyl) triethoxysilane, reaction carries out Magneto separate after 2 days, and is replaced with second alcohol and water
Washing respectively obtains the magnetic microsphere 2 of amido modified Silica-coated afterwards twice.
(4) by the magnetic microsphere 2 of 10mg step (3) the amido modified Silica-coated and 4mg hyalomitome acid-mixed
It closes, adds 10mg EDC and 5mg NHS, reaction obtained the magnetic microsphere 2 of the high density carboxyl modified after 2 hours.
Embodiment 3
(1) in air, by 50gFeCl2·H2O is added in 100mL deionized water, and obtaining concentration is 0.50g/mL's
FeCl2Aqueous solution.To 100mLFeCl2·H2Ammonium hydroxide 50mL is added in O aqueous solution, after stirring 90min, color gradually becomes light green
Color, then green is deepened, eventually become black；
(2) 5g oleic acid is added into step (1), after mixing sets mixed liquor in closed reaction kettle, at 130 DEG C
Lower heating is reacted 3 hours, is then alternately washed respectively twice, is dispersed in normal octane after Magneto separate, i.e., with deionized water and ethyl alcohol
The paramagnetic nanoparticles Fe of the oleic acid package of black can be obtained3O4 3。
(3) the paramagnetic nanoparticles Fe of Xiang Hanyou 10mg oleic acid package3O4125mg ammonium hydroxide, the positive silicon of 40mg are added in 3 solution
Sour tetra-ethyl ester and 5mg (3- aminopropyl) triethoxysilane, reaction carries out Magneto separate after 3 days, and is replaced with second alcohol and water
Washing respectively obtains the magnetic microsphere 3 of amido modified Silica-coated afterwards twice.
(4) by the magnetic microsphere 3 of 10mg step (3) the amido modified Silica-coated and 8mg polyglutamic acid-mixed
It closes, adds 40mg EDC and 20mg NHS, reaction obtained the magnetic microsphere 3 of the high density carboxyl modified after 3 hours.
Embodiment 4
10mg polyacrylic acid is substituted for 20mg succinamic acid dendroid respectively and gathered by method in accordance with the above-mentioned embodiment 1
Close object, the magnetic microsphere 4 of available high density carboxyl modified.
The magnetic microsphere 1 of amido modified Silica-coated in embodiment 1 is scanned electron microscope (SEM)
Detection and transmission electron microscope (TEM) detection, as a result as depicted in figs. 1 and 2.As can be seen that amido modified silica
The size of the magnetic microsphere 1 of package is uniform, narrow size distribution, and good dispersion.Meanwhile kernel is coated with multiple magnetic Nanos
Particle, outer shell boundary is obvious, illustrates that stability is good, not easy to reunite.The amido modified Silica-coated of other embodiments
Magnetic microsphere SEM and TEM detection there is similar effect, do not repeat one by one.
5 extraction of plasmid DNA of embodiment
1.5mL E. coli broth is poured into centrifuge tube, 12000rpm is centrifuged 30s and abandons supernatant collection thallus, to thallus
Buffer (50mM glucose, 25mM Tris-HCl (pH8.0), 10mM EDTA that 100 μ L contain 20 μ g/mL RNA enzyme is added
(pH8.0)), 200 μ L alkaline lysis liquid (SDS of NaOH containing 0.2M and 1%), soft to mix, ice bath 3min, then plus 150 μ L pH
The 5M liquor kalii acetici that value is 5, mixing are stood, and 12000rpm is centrifuged 10 minutes, supernatant are transferred in new centrifuge tube；To
The magnetic microsphere 1.5mg of the high density carboxyl modified of embodiment 1 is added in centrifuge tube, concussion mixes, and incubates 2-5min, will be centrifuged
Pipe is placed on Magneto separate frame, is inhaled and is abandoned supernatant；70% ethyl alcohol of 500 μ L is added to clean Plasmid DNA-magnetic particle compound twice,
Ethyl alcohol is drained, and TE buffer (10mM Tris-HCl (pH8.0), 1mM EDTA (pH8.0)) suspended magnetic particle of 50 μ L is added,
Magneto separate collects eluent.Ultraviolet detection, DNA have ultraviolet characteristic absorption peak at 260nm, and the purity of DNA is with (OD260-
OD320)/(OD280-OD320) determine, which should be between 1.7-1.9.OD260The extinction for being sample solution at 260nm
Value, OD280The light absorption value for being sample solution at 280nm, OD320Background value when for sample solution ultraviolet determination.The result shows that
The Plasmid DNA of about 40 μ g high-purities is obtained, purity is 1.8 or so.1% Ago-Gel of concentration is prepared, the matter of said extracted is taken
Grain DNA carries out electrophoresis, and electrophoresis result is observed on gel imaging system after 25min.
Electrophoresis result fig. 3, it is shown that high density carboxyl magnetic bead extract Plasmid DNA it is very complete, extract result
Differences between batches are small, yield and purity is high.
After taking the magnetic microsphere 1~4 of the high density carboxyl modified in 1mg above-described embodiment 1~4 to activate, it is added 200 μ g's
BSA is incubated for 12 hours at 4 DEG C after mixing.Then 3 are washed with the PBS buffer solution (Tween containing 0.1%) that pH is 7.4
It is secondary, obtain immunomagnetic beads 1~4.And take commercially available carboxyl modified silica (Magnetic bead) package magnetism it is micro-
Ball is as a control group.
BSA detection in surface is carried out to above-mentioned immunomagnetic beads 1~4, obtains that the results are shown in Table 1:
| Every milligram of difference BSA package amount | |
| Embodiment 1 | 80 micrograms |
| Embodiment 2 | 76 micrograms |
| Embodiment 3 | 110 micrograms |
| Embodiment 4 | 102 micrograms |
| Control group | 20 micrograms |
The BSA package amount of 1 immunomagnetic beads of table
As it can be seen from table 1 being exempted from the magnetic microsphere of high density carboxyl modified of the invention and biological ligand cou
Epidemic disease magnetic bead substantially increases the BSA package amount on immunomagnetic beads surface.
For those skilled in the art, without departing from the principles of the embodiments of the present invention, also
Several improvements and modifications can be made, these improvements and modifications are also considered as the protection scope of the embodiment of the present invention.
Claims (4)
1. a kind of preparation method of the magnetic microsphere of high density carboxyl modified, it is characterised in that: the preparation method includes following
Step:
S1: amido modified magnetic microsphere is prepared；
S2: the amido modified magnetic microsphere is mixed with organic carboxyl acid, adds EDC and NHS, after reaction 1~3 hour
To the magnetic microsphere of the high density carboxyl modified；
In the step (2), the organic carboxyl acid is polyacrylic acid, hyaluronic acid, polyglutamic acid or succinamic acid dendroid
Polymer；
In the step (2), EDC, NHS of addition, organic carboxyl acid, amido modified magnetic microsphere mass ratio be (10~
60): (5~30): (4~20): 10；
Wherein, in the step s1, amido modified magnetic microsphere is the magnetic microsphere of amido modified Silica-coated, it
It is prepared via a method which to obtain: ammonium hydroxide, silylating reagent and amino being added in the solution of Xiang Hanyou magnetic nano-particle
Silane coupling agent, reaction obtained the magnetic microsphere of amido modified Silica-coated after 1~3 day；The magnetic nano-particle,
The mass ratio that ammonium hydroxide, silylating reagent, amino silicane coupling agent are added are as follows: 1:(12.5~40): (2~8): (0.5~3)；Ammonia
The mass concentration of water is 25-28%；
The magnetic nano-particle is magnetic Nano Fe3O4, it is to be made by the steps to obtain:
S1-1: in air, ammonium hydroxide being added into the aqueous solution of divalent iron salt, and it is ferric iron that stirring, which keeps part ferrous oxidising,
Oxidation precipitation generates black Fe under alkaline condition3O4Particle；
S1-2: oleic acid being added into step s1-1, and heating reaction 3~5 hours at 60~130 DEG C, can obtain after mixing
The magnetic Nano Fe wrapped up to the oleic acid3O4 。
2. the magnetic microsphere for the high density carboxyl modified that one kind is prepared method according to claim 1.
3. a kind of nucleic acid extraction kit, including magnetic microsphere, it is characterised in that: the magnetic microsphere is claim 2
The magnetic microsphere of the high density carboxyl modified.
4. a kind of immunomagnetic beads, it is characterised in that: the immunomagnetic beads are by high density carboxyl modified described in claim 2
Magnetic microsphere and biological ligand cou, are prepared.
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| CN103990423A (en) * | 2014-03-27 | 2014-08-20 | 华南师范大学 | Single-stranded DNA aptamer modified SiO2/Fe3O4 magnetic microsphere preparation method |
| CN104893727A (en) * | 2015-04-23 | 2015-09-09 | 天津大学 | Carboxyl functionalized micro-scale rod-like upconversion fluorescence material and preparation method thereof |
| CN105771904A (en) * | 2016-03-09 | 2016-07-20 | 清华大学 | Magnetic adsorbent as well as preparation method thereof, and recycling of palladium in nitric acid medium |
| CN105664887A (en) * | 2016-03-14 | 2016-06-15 | 中国地质大学（武汉） | Preparation method of functional magnetic silicon balls |
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Inventor after: Cai Hongdong Inventor after: Zhang Xianglin Inventor after: Liu Guan Inventor after: Chen Changyue Inventor before: Liu Guan Inventor before: Cai Hongdong Inventor before: Chen Changyue Inventor before: Zhang Xianglin |