CN110196324A - Magnetic mesoporous Nano particles of silicon dioxide probe and its preparation method and application - Google Patents
Magnetic mesoporous Nano particles of silicon dioxide probe and its preparation method and application Download PDFInfo
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- CN110196324A CN110196324A CN201910475019.4A CN201910475019A CN110196324A CN 110196324 A CN110196324 A CN 110196324A CN 201910475019 A CN201910475019 A CN 201910475019A CN 110196324 A CN110196324 A CN 110196324A
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Abstract
The invention discloses magnetic mesoporous Nano particles of silicon dioxide probes, the magnetic mesoporous Nano particles of silicon dioxide probe is that magnetic mesoporous nano SiO 2 particle loading peroxidase is first obtained M-MSN/POD, polyethyleneimine package M-MSN/POD encapsulation peroxidase is obtained into M-MSN/POD/PEI nanoparticle again, then is mixed to form magnetic mesoporous Nano particles of silicon dioxide probe with antibody.The invention also discloses the preparation methods of magnetic mesoporous Nano particles of silicon dioxide probe.The probe can be used for cell targeted specific detection, the Visual retrieval of virus, prepare overdelicate test strip, compared to the prior art, have specificity high with products of the present invention, stability is good, the features such as application field is extensive, in addition simple process, cost is relatively low, therefore has good promotion prospect.
Description
Technical field
The invention belongs to meso-porous titanium dioxide silicon probe preparation technical fields, and in particular to magnetic mesoporous silica dioxide nano particle
Sub- probe and its preparation method and application.
Background technique
Since Kresge in 1992 et al. reports a series of material of New Mesoporous Molecular Sieves for the first time, mesoporous two
Silica material has just caused the extensive concern of researcher.Mesoporous silicon oxide is simply and low in cost due to synthesizing, and has big
Specific surface area, size and the adjustable vesicular texture of structure and be easy to chemical modification surfaces externally and internally make they become dress
Carry the excellent carrier of small molecule.
In past a period of time, people are to Metaporous silicon dioxide material in absorption, imaging, drug delivery and biology
A series of research has been carried out in the application of sensor etc..Mesoporous silicon oxide has porous performance, general in the course of the research
All over for carrying other small-molecule substances such as drug, albumen, fluorescent material.With preferable delivered payload capability and biodegradable
Performance.Huge application potential is shown in biological diagnosis analysis field, and achieves fruitful research achievement.Mesoporous shell
Layer imparts two surfaces M-MSNs, first is that entire particle surface, second is that mesoporous inner surface.In this way, the duct of mesoporous material is not
It is only capable of having an effect on the surface of the material with the comparable biomolecule of scale, and this effect runs through the entire ordered arrangement of material
Jie see limitation space in, and then promote guest molecule controlled release.Secondly, mesoporous lamella again can be with effective protection inside
Magnetic nano-particle, solve the disadvantage that magnetic nano-particle itself it is unstable, it is easy to reunite and oxidation, make M-MSNs have more
The advantage of advantageous composite synergistic.Again, said from multi-functional compound angle, on the hole wall of mesopore silicon oxide layer can flexibly and
Functional modification is easily realized again, so as to optimize the interaction between tote-carrier, control surface charge, dispersion
Stability and targeting.
The means for clinically detecting virosis are most of more complicated, and laboratory testing has high operation requirements, need specially
The disadvantages of operation of industry personnel and time-consuming, expensive equipment.The signal amplification effect being often used when laboratory testing is most typical
Test is enzyme-linked immunosorbent assay, can use chemical means and realizes that signal amplifies, but enzyme-linked immunosorbent assay
Operating process is complicated, and professional is needed to operate, and operating process error easily occurs and causes result error.
Summary of the invention
Goal of the invention:, the present invention quasi- signal amplification principle using ELISA of the existing technology in order to solve the problems, such as,
Peroxidase is carried using magnetic mesoporous Nano particles of silicon dioxide, it can be with the principle of layer assembly, group using positive and negative charge
It dresses up as nano-probe, it can virion directly in test sample.The high catalytic efficiency of enzyme is utilized, can greatly amplify anti-
Effect is answered, to make measuring method reach very high susceptibility, while the instrument that can reduce ELISA valuableness again uses, and is facing
There is apparent advantage in the detection of bed.
Technical solution: in order to solve the above-mentioned technical problem, the technical scheme adopted by the invention is as follows: one kind magnetic mesoporous two
Silicon oxide nanoparticle probe, the magnetic mesoporous Nano particles of silicon dioxide probe are first by magnetic mesoporous silica nanometer
Particle loads peroxidase and obtains M-MSN/POD, then polyethyleneimine package M-MSN/POD encapsulation peroxidase is obtained
M-MSN/POD/PEI nanoparticle, then magnetic mesoporous Nano particles of silicon dioxide probe is mixed to form with antibody.
Wherein, magnetic mesoporous nano SiO 2 particle is amido modified, size 60-65nm, mesoporous pore size 5-
6nm。
Wherein, the polyethyleneimine is linear, molecular weight 25KDa.
The content of present invention further includes the preparation method of the magnetic mesoporous Nano particles of silicon dioxide probe, including following
Step:
1) loading of peroxidase: peroxidase and M-MSN nano particle are mixed, and obtain the M-MSN for loading POD
Nano particle, as M-MSN/POD；
2) PEI wraps up M-MSN-/POD: PEI being mixed with M-MSN/POD, obtains the magnetism that polyethyleneimine package loads
Nanometer particle, that is, M-MSN/POD/PEI nanoparticle；
3) coupling of antiviral antibody: M-MSN/POD/PEI nanoparticle and antibody are mixed, and products therefrom mixing obtains
Magnetic mesoporous Nano particles of silicon dioxide probe.
Wherein, the mass ratio of the peroxidase in the step 1) and M-MSN nano particle is 1: 1.
Wherein, the mass ratio of the step 2) PEI and M-MSN/POD is 1: 10.
Wherein, the step 2) incorporation time is 8-12h.
Wherein, step 3) the M-MSN/POD/PEI nanoparticle and antiviral antibody mass ratio are 10: 1.
Wherein, the antibody of the step 3) includes but are not limited to newcastle disease virus antibody, other antibody can be applicable in.
Wherein, the virus includes but are not limited to newcastle disease (NDV) virus, other viruses can be applicable in.
The content of present invention further includes that magnetic mesoporous Nano particles of silicon dioxide probe detects in viral test strips in preparation
Using.
The utility model has the advantages that compared with prior art, the invention has the advantages that
1) present invention utilizes the carrier band features of porous structure, have carried common chemo-responsive dyes peroxidase, benefit
With the mesoporous characteristic of mesoporous silicon oxide, peroxidase is loaded into meso-hole structure, and poly- using high molecular polymer
Aziridine ensure that the enzymatic activity of mesoporous middle peroxidase with temporary " closure " mesoporous aperture.The preparation method combines
The signal amplification principle of traditional enzyme-linked immunosorbent assay, may finally carry out instruction testing result in a manner of colour developing；
The specific detection that the novel nano probe prepared in this way can be used for having cell targeted, viral visualization inspection
It surveys, prepares overdelicate test strip, compared to the prior art, have specificity high with products of the present invention, stablize
The features such as property is good, and application field is extensive, simple process in addition, cost is relatively low, therefore has good promotion prospect.
2) reaction condition of the present invention is very mild, will not influence and protein active, the dispersion degree of the probe in the solution
Also very well, the specific detection for the treatment of and cause of disease to some diseases such as cancers provides technical strategies and research is thought
Road.
Detailed description of the invention
The TEM phenogram of the magnetic mesoporous silica of Fig. 1；
The SEM phenogram of the magnetic mesoporous silica of Fig. 2；
The magnetic mesoporous silica of Fig. 3 loads the SDS-PAGE figure of peroxidase；Wherein 1 is pre-dyed protein labeling, and 2 are
Probe magnetic frame supernatant, 3 load peroxidase for magnetic mesoporous silica, and 4 be magnetic mesoporous silica, and 5 be simple
20 μ g of peroxidase；
SDS-PAGE figure of the evidence figure verifying nanometer particle of Fig. 4 probe preparation in conjunction with antibody；Wherein M is pre-
Protein labeling is contaminated, 1 is the peroxidase of 2 μ g, and 2 be the antibody of 2 μ g, and 3 load peroxidase and antibody for the magnetic empty silicon that is situated between
The nano-probe of link；
The pcr amplification product of Fig. 5 embodiment 3 carries out nucleic acid gel electrophoretogram, and swimming lane M is marker2000, and swimming lane I is
NDV virus；Swimming lane II is POD/M-MSN/PEI/Ab/NDV；
The pcr amplification product of Fig. 6 embodiment 4 carries out nucleic acid gel electrophoretogram, and swimming lane M is marker2000, swimming lane INDV
Virus；Swimming lane II is NDV: POD/M-MSN/PEI/Ab=1: 1；Swimming lane III is NDV:POD/M-MSN/PEI/Ab=1/2；Swimming
Road IV is NDV:POD/M-MSN/PEI/Ab=1/4；Swimming lane V is NDV:POD/M-MSN/PEI/Ab=1/6；
The TEM of the magnetic mesoporous silica nanometer probe combination NDV virus of Fig. 7 combines figure；
The pcr amplification product of Fig. 8 embodiment 6 carries out nucleic acid gel electrophoretogram, and swimming lane M is marker2000, and swimming lane I is
H9N2 virus；Swimming lane II is POD/M-MSN/PEI/Ab/H9N2；
TEM figure in Fig. 9 embodiment 7.
Specific embodiment
Below by specific embodiment, the present invention is further described, it is noted that for the ordinary skill of this field
For personnel, without departing from the principle of the present invention, several variations and modifications can also be made, these also should be regarded as belonging to
Protection scope of the present invention.Experimental method in following embodiments is unless otherwise specified conventional method.Following embodiments
Used in experimental material be unless otherwise specified to be commercially available from routine biochemistry reagent shop.In following embodiment
Quantitative experiment is respectively provided with three repeated experiments, and results are averaged.
Magnetic mesoporous nano SiO 2 particle is purchased from Shanghai Suo Fei biotech firm, article No. are as follows: M-MSNs-50；Peroxidating
Object enzyme is purchased from Beijing Suo Laibao biotech firm；Newcastle disease virus (strain: lasota) Yangzhou University, Jiangsu Province veterinary college is natural
Immune signal, immune and infection research room are that laboratory saves strain；Newcastle disease virus antibody is Newcastle Disease
Virus antibody | 8H2 (BIO-RAD, MCA2822)；Magnetic frame machine is purchased from Eppendorf company, Germany；PBS (10X) is adopted
Purchased from the green skies Bioisystech Co., Ltd in Jiangsu；Super-clean bench, blending instrument are purchased from Thermo Fisher company；RNA virus mentions
Take kit buying from Beijing Yuanpinghao Biological Technology Co., Ltd.；RNA reverse transcription reagent box is century biology purchased from Beijing health
Science and Technology Ltd.；2 × Taq Master Mix (p111-w3) is bought from Nanjing Vazyme Biotechnology Co., Ltd.；Primer
Synthesis comes from Sangon Biotech (Shanghai) Co., Ltd., Yangzhou University, the source Jiangsu Province veterinary science of influenza virus H9N2
Institute's innate immunity signal, immune and infection research room are that laboratory saves strain.The PBS used in following all examples is
1 × PBS (PH7.2-7.4) after dilution.
Embodiment 1 constructs magnetic mesoporous Nano particles of silicon dioxide probe by taking newcastle disease virus detection probe as an example
1, the characterization of M-MSN nano particle: being diluted to 1 μ g/ μ L for M-MSN nano particle, then take 10 μ L point sample copper mesh,
Then the size and aperture, the result is shown in Figure 1 and Fig. 2 of transmission electron microscope and scanning electron microscopic observation nanoparticle.
2, the loading of peroxidase (POD): by the M-MSN after the dilution in the peroxidase and step 1 of commercialization
Nano particle according to mass ratio 1: 1 (i.e. the POD of M-MSN: the 100 μ g of 100 μ g) mix, mixed solution 4 DEG C on mixture,
It is incubated for 8-12h；Then the unbonded peroxidase of magnetic frame separating, washing, obtains supernatant precipitating, and obtained precipitating is
POD/M-MSN；Precipitating is dissolved in the PBS of 100 μ L, it is spare；The M-MSN nano particle of 20 μ g is taken to be dissolved in 100 μ L respectively simultaneously
PBS in, the POD of 20 μ g is dissolved in the PBS of 20 μ L, spare；
20 μ L of precipitating after taking 20 μ L of supernatant respectively, being dissolved in PBS, 20 μ L of M-MSN nano particle for being dissolved in PBS, it is dissolved in PBS
20 μ L of POD, be added 5 μ L 5 × loading buffer, 100 DEG C are boiled sample 5 minutes, then do proteins gel electrophoresis, voltage
120V, electric current 90mA.Then time 1h uses coomassie brilliant blue staining 25min, destainer decolourizes for 24 hours, and the results are shown in attached figure 3.From
In Fig. 3 as can be seen that POD/M-MSN in really by the POD of script load to M-MSN it is mesoporous in.
3, it is mixed with 5% polyethyleneimine according to mass ratio 10: 1 with the M-MSN/POD that step 2 constructs, 4 DEG C,
It is incubated for 1h；Magnetic frame carries out separation supernatant and precipitating, is precipitated as M-MSN/POD/PEI nanoparticle, spare.
4, the coupling of newcastle disease virus antibody: by the M-MSN/POD/PEI nanoparticle and newcastle disease virus antibody of step 3
Newcastle Disease Virus antibody | 8H2 (BIO-RAD, MCA2822) is mixed according to 10: 1 ratio, and 4 DEG C,
It is incubated for 8h；Magnetic mesoporous Nano particles of silicon dioxide probe POD/M-MSN/PEI/Ab is obtained, POD/M-MSN/PEI/Ab is molten
It is spare in the PBS of 20 μ L；2 μ g of antibody is dissolved in the PBS of 20 μ L simultaneously；The POD of 2 μ g is dissolved in the PBS of 20 μ L；It is spare.
20 μ L of the POD for being dissolved in PBS is taken respectively, 20 μ L of newcastle disease virus antibody that is dissolved in PBS, is dissolved in POD/M- in PBS
20 μ L of MSN/PEI/Ab be added 5 μ L 5 × loading buffer.100 DEG C, boil sample 5min.Then protein electrophoresis is done, electricity
Press 120V, electric current 90mA.Time 1h, then uses coomassie brilliant blue staining 25min, and destainer decolourizes for 24 hours.It takes pictures, as a result sees
Attached drawing 4, it can be seen from the figure that newcastle disease virus antibody with POD/M-MSN/PEI successful connection.
The detection of 2 peroxidase activity of embodiment
In order to detect the activity of the peroxidase in magnetic mesoporous silicon oxide, we select using TEM to be substrate, with
Peroxidase under different conditions carries out reaction solution, is specifically grouped into simple peroxidase (POD), magnetic mesoporous two
Peroxidase (POD/M-MSN), the polyethyleneimine package of silicon oxide-wrapped are loaded with magnetic mesoporous the two of peroxidase
The nano-probe (POD/M-MSN/PEI/Ab) that silica (POD/M-MSN/PEI) and building are completed, is then read using microplate reader
Take the numerical value of its OD450.After peroxidase is loaded into magnetic mesoporous silica by testing result discovery, in each shape
The enzymatic activity of peroxidase does not have any change under state.Specific testing result is as follows:
| The content of POD | OD450 |
| POD 0.2μg | 3.9267 |
| POD/M-MSN 0.2μg | 3.9389 |
| POD/M-MSN/PEI 0.2μg | 3.9165 |
| POD/M-MSN/PEI/Ab 0.2μg | 3.7982 |
The magnetic mesoporous Nano particles of silicon dioxide probe in detecting newcastle disease virus of embodiment 3
1, the magnetic mesoporous Nano particles of silicon dioxide probe (POD/M-MSN/PEI/Ab) prepared embodiment 1 and new city
Epidemic disease poison combines: magnetic mesoporous Nano particles of silicon dioxide probe ratio in conjunction with newcastle disease virus is 1: 1, and room temperature reaction 1 is small
When, using magnetic frame separation supernatant and precipitating after 1 hour, cleaned 3 times with 1 × PBS.It precipitates spare.
2, viral RNA and reverse transcription are extracted: gained precipitated product in step 1 is extracted into RNA using RNA extracts kit,
It then the use of reverse transcription reagent box is cDNA by the RNA reverse transcription extracted.Reverse transcription system is as follows:
Transcriptive process,reversed is 37 DEG C, 15min, 85 DEG C, 5s.
3, joint efficiency detects: the reverse transcription product of step 2 being carried out PCR identification, magnetic mesoporous silicon oxide is detected and receives
The joint efficiency of rice probe.According to the complete genome sequence (GenBank accession no.:AE001363) of newcastle disease virus,
Utilize the specific primer of the 5.0 software design gene of Primier, upstream primer (F): 5 '-
ctatccgggttgcgctggta-3′(SEQ ID No.1)；Downstream primer (R): 5 '-ttccaagtaggtggcacgca-3 '
(SEQ ID No.2), the length of amplified production are 866bp.The reaction system of PCR identification is specific as follows:
Prepare the PCR reaction system that total volume is 25 μ L
PCR response procedures: 95 DEG C of 5min, 95 DEG C of 5s, 57 DEG C of 30s, 72 DEG C of 45s, 72 DEG C of 10min amount to 25 and recycle, and 13
℃50min。
Pcr amplification product carries out agarose gel electrophoresis: configuring 1.5% Ago-Gel, electrophoretic parameters are voltage
120V, time 40min.Electrophoresis result is referring to Fig. 5, the results show that the efficiency of nanometer probe combination newcastle disease virus
It is higher, newcastle disease virus is not detected in the supernatant after the separation of nanometer particle probe.
The sensitivity test of the magnetic mesoporous Nano particles of silicon dioxide probe in detecting newcastle disease virus of embodiment 4
1, the magnetic mesoporous Nano particles of silicon dioxide probe (POD/M-MSN/PEI/Ab) for preparing embodiment 1 from it is different
The newcastle disease virus of ratio is combined, and keeps magnetic mesoporous Nano particles of silicon dioxide probe (POD/M-MSN/PEI/Ab)
Measure it is constant in the case where, reduce virus amount；Specific choice ratio is as follows: NDV: POD/M-MSN/PEI/Ab=1: 1；NDV:
POD/M-MSN/PEI/Ab=1/2；NDV:POD/M-MSN/PEI/Ab=1/4；NDV:POD/M-MSN/PEI/Ab=1/6, room
Using magnetic frame separation supernatant and precipitating after temperature reaction 1 hour, 1 hour, cleaned 3 times with 1 × PBS.It precipitates spare.
2, viral RNA and reverse transcription are extracted: gained precipitated product in step 1 is extracted into RNA using RNA extracts kit,
It then the use of reverse transcription reagent box is cDNA by the RNA reverse transcription extracted.Reverse transcription system is as follows:
Transcriptive process,reversed is 37 DEG C, 15min, 85 DEG C, 5s.
3, joint efficiency detects: the reverse transcription product of step 2 being carried out PCR identification, magnetic mesoporous silicon oxide is detected and receives
The joint efficiency of rice probe.According to the complete genome sequence (GenBank accession no.:AE001363) of newcastle disease virus,
Utilize the specific primer of the 5.0 software design gene of Primier, upstream primer (F): 5 '-
ctatccgggttgcgctggta-3′(SEQ ID No.1)；Downstream primer (R): 5 '-ttccaagtaggtggcacgca-3 '
(SEQ ID No.2), the length of amplified production are 866bp.The reaction system of PCR identification is specific as follows:
Prepare the PCR reaction system that total volume is 25 μ L
PCR response procedures: 95 DEG C of 5min, 95 DEG C of 5s, 57 DEG C of 30s, 72 DEG C of 45s, 72 DEG C of 10min amount to 25 and recycle, and 13
℃50min。
Pcr amplification product carries out agarose gel electrophoresis: configuring 1.5% Ago-Gel, electrophoretic parameters are voltage
120V, time 40min.Electrophoresis result is referring to Fig. 6, the results show that the efficiency of nanometer probe combination newcastle disease virus
It is higher, when the amount of virus is reduced to 1/6, it still is able to capture newcastle disease virus.
Detection of the magnetic mesoporous Nano particles of silicon dioxide probe of embodiment 5 to newcastle disease virus
By probe POD/M-MSN/PEI/Ab in conjunction with newcastle disease virus, reacts at room temperature the time 1 hour, then use magnetic force
Frame separates the virus being not associated in supernatant, is cleaned probe 3 times using PBS.Then mesoporous silicon dioxide nano probe and new city are taken
The product that epidemic disease poison combines, point drying at room temperature 10 minutes, then use 2% phosphotungstic acid negative staining in copper mesh, and the time is 5 minutes,
Copper mesh is placed in electric microscopic observation, as shown in fig. 7, magnetic mesoporous silica nanometer probe is in conjunction with newcastle disease virus.
Embodiment 6
For the specificity of detection probe, we are by the POD/M-MSN/PEI/Ab nano-probe constructed in embodiment 3 and flow
Influenza Virus H9N2 is combined, and the room temperature reaction time is similarly 1 hour, is separated after reaction using magnetic frame, and PBS is cleaned three times,
Then products therefrom is extracted into RNA, it is cDNA that gained RNA, which is carried out reverse transcription, and reverse transcription system is as follows:
Transcriptive process,reversed is 37 DEG C, 15min, 85 DEG C, 5s.
Then PCR detects joint efficiency.
Prepare the PCR reaction system that total volume is 25 μ L
PCR response procedures: 95 DEG C of 5min, 95 DEG C of 5s, 57 DEG C of 30s, 72 DEG C of 45s, 72 DEG C of 10min amount to 25 and recycle, and 13
℃50min。
Pcr amplification product carries out agarose gel electrophoresis: configuring 1.5% Ago-Gel, electrophoretic parameters are voltage
120V, time 40min.As a result referring to Fig. 8, do not occur any band.It can prove that the probe has specificity, eliminate non-
The possibility of specific adsorption.
Embodiment 7
For the specificity of detection probe, we are by the nano-probe and stream of the POD/M-MSN/PEI/Ab in embodiment 3
Influenza Virus H9N2 is combined, and the room temperature reaction time is similarly 1 hour, is separated after reaction using magnetic frame, and PBS is cleaned three times；
Then product of the mesoporous silicon dioxide nano probe in conjunction with virus H9N2 is taken, puts in copper mesh, drying at room temperature 10 minutes, then makes
With 2% phosphotungstic acid negative staining, the time is 5 minutes, copper mesh is placed in electric microscopic observation, concrete outcome is shown in Fig. 9, does not find mesoporous two
The electromicroscopic photograph that silica nano-probe is combined with virus.
In conclusion mesoporous silicon dioxide nano probe prepared by the present invention has specificity well, the later period has very
Big potentiality to be exploited can be applied in other detection fields such as detection, the detection chips of test strips.
It above are only the preferred embodiment of the invention, be not restricted to the present invention.Those skilled in the art is come
It says, other various forms of variations or variation can also be made on the basis of the above description.There is no need and unable to all
Embodiment illustrate.And the obvious changes or variations that thus scheme is extended out are still in protection of the invention
Within the scope of.
Sequence table
<110>Yangzhou University
<120>magnetic mesoporous Nano particles of silicon dioxide probe and its preparation method and application
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
ctatccgggt tgcgctggta 20
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<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
ttccaagtag gtggcacgca 20
Claims (10)
1. a kind of magnetic mesoporous Nano particles of silicon dioxide probe, which is characterized in that the magnetic mesoporous silica dioxide nano particle
Sub- probe is that magnetic mesoporous nano SiO 2 particle loading peroxidase is first obtained M-MSN/POD, then by polyethyleneimine
Amine package M-MSN/POD encapsulation peroxidase obtains M-MSN/POD/PEI nanoparticle, then is mixed to form magnetic Jie with antibody
Hole Nano particles of silicon dioxide probe.
2. magnetic mesoporous Nano particles of silicon dioxide probe according to claim 1, which is characterized in that magnetic mesoporous dioxy
SiClx nano particle is amido modified, size 60-65nm, mesoporous pore size 5-6nm.
3. magnetic mesoporous Nano particles of silicon dioxide probe according to claim 1, which is characterized in that the polyethyleneimine
Amine is linear, molecular weight 25KDa.
4. the preparation method of magnetic mesoporous Nano particles of silicon dioxide probe described in claim 1, which is characterized in that including with
Lower step:
1) loading of peroxidase: peroxidase and M-MSN nano particle are mixed, and obtain the M-MSN nanometer for loading POD
Particle, as M-MSN/POD；
2) PEI wraps up M-MSN-/POD: PEI being mixed with M-MSN/POD, obtains the magnetic mesoporous of polyethyleneimine package loading
Silicon nano, that is, M-MSN/POD/PEI nanoparticle；
3) coupling of antibody: M-MSN/POD/PEI nanoparticle and antibody are mixed, and products therefrom mixing obtains magnetic mesoporous
Nano particles of silicon dioxide probe.
5. according to right ask 4 described in magnetic mesoporous Nano particles of silicon dioxide probe preparation method, which is characterized in that it is described
The mass ratio of peroxidase and M-MSN nano particle in step 1) is 1:1.
6. according to right ask 4 described in magnetic mesoporous Nano particles of silicon dioxide probe preparation method, which is characterized in that it is described
The mass ratio of step 2 PEI and M-MSN/POD are 1:10.
7. according to right ask 4 described in magnetic mesoporous Nano particles of silicon dioxide probe preparation method, which is characterized in that it is described
Step 2 incorporation time is 8-12h.
8. according to right ask 4 described in magnetic mesoporous Nano particles of silicon dioxide probe preparation method, which is characterized in that it is described
Step 3) M-MSN/POD/PEI nanoparticle and antibody mass ratio are 10:1.
9. the preparation method of magnetic mesoporous Nano particles of silicon dioxide probe according to claim 4, which is characterized in that institute
The antibody for stating step 3) is newcastle disease virus antibody.
10. the described in any item magnetic mesoporous Nano particles of silicon dioxide probes of claim 1 ~ 3 detect viral test paper in preparation
Application in item.
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| CN103575892A (en) * | 2012-08-08 | 2014-02-12 | 中国科学院电子学研究所 | Kit for quantitatively detecting CEA (Carcino-Embryonic Antigen) as well as preparation method and application method of kit |
| CN102989398A (en) * | 2012-12-02 | 2013-03-27 | 复旦大学 | Magnetic inorganic nano particle/large-aperture ordered mesopore oxide nuclear shell microspheres and preparation method thereof |
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Inventor after: Ni Chang Inventor after: Zhou Xin Inventor before: Zhou Xin Inventor before: Li Zhenzhen |