CN104360064A - Monodisperse hollow gold nanosphere and preparation method and application thereof - Google Patents
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Abstract
The invention discloses a monodisperse hollow gold nanosphere which is provided with a monodisperse hollow kernel and a monodisperse gold nano outer layer, wherein the monodisperse gold nano outer layer is one or more of a gold nano layer, a silver nano layer, a copper nano layer and a platinum nano layer; the monodisperse hollow kernel is formed by eroding a monodisperse organic or inorganic nanosphere. Compared with a conventional hollow gold nanosphere, the hollow gold nanosphere disclosed by the invention is high in monodispersity. Therefore, an SERS (surface enhanced raman scattering) probe prepared by the hollow gold nanosphere with high monodispersity has immunoassay characteristics of extremely high uniformity, sensitivity and repetitiveness. High-precision quantitative detection is facilitated during quantitative analysis of biological markers.
Description
Technical field
The present invention relates to a kind of technical field strengthening raman material and prepare, more specifically, relate to a kind of monodispersity hollow gold nanometer ball and its preparation method and application.
Background technology
In the immunoassay Study and appliance detected based on Surface enhanced raman spectroscopy (SERS), hollow metal nanometer is commonly used for the probe of SERS mark, builds the sandwich immunoassays of high sensitivity, homogeneity and signal repeatability.For the super sensitivity detection of common tumor markers, much useful diagnostic data can be provided.The hollow metal nanosphere of previous preparation is all be core by cobalt nanosphere usually, and at its superficial growth gold shell; And then remove cobalt nanometer core, obtain hollow metal nano ball.But, because current cobalt nanosphere technology of preparing can't realize the preparation of monodispersity cobalt nanosphere, in this way can not obtain the hollow gold nanometer ball of homogeneity, this has impact to the quantitative test of follow-up sandwich immunoassays.
At most developing countries, tumour (i.e. cancer) of feeling sick is the disease that second largest mortality ratio is the highest, and it have impact on human body more than 60 human organs.Cancer known at present has kind more than 200, and wherein lung cancer is that death toll is maximum, accounts for 1/4 of whole cancer.In order to cancer patient improves survival rate and prognosis evaluation, early stage clinical diagnosis room is extremely important and necessary.Such as, but the early stage diagnosis of lung cancer encounters many challenges, because lung cancer does not generally have obvious specific symptoms in early days, coughs, breathes hard, become thin.In tensth century in past, many technology are developed and for cancer detection, comprise CT, NMR, chest x-ray, PET-CT etc.But they also have very large shortcoming: expensive, time-consuming, muting sensitivity.
Biomarker is the index of cell physiological state and changes along with the generation development of disease.Serum biomarkers is all very useful in early diagnosis of tumor and assessment result for the treatment of.The technology being conventionally used to antigen and antibody has RIA, FIA, ELISA, ECIA etc.Among them, fluoroimmunoassay is widely used in bioanalysis due to its high sensitivity and low detection limit.But its Broad excitation band limits it and uses.
Summary of the invention
Goal of the invention: first object of the present invention is to provide a kind of monodispersity hollow metal nanosphere, has the immunoassay characteristic of very good homogeneity, sensitivity and repeatability as SERS probe material.Second object of the present invention is to provide the preparation method of above-mentioned monodispersity hollow metal nanosphere.3rd object of the present invention is to provide the application of above-mentioned monodispersity hollow metal nanosphere in preparation SERS probe.
In the present invention, first, with or organic nanosphere inorganic by monodispersity for core, then plated metal forms the good metal nano shell of monodispersity on this core, and namely the core finally removing metal nano shell obtain hollow metal nano ball.The nanosphere technology of preparing of current organic or inorganic is ripe, and the nanosphere monodispersity of preparation is good.The present invention selects the good nanosphere of the monodispersity of organic or inorganic to be core, the gold nanoshell that preparation monodispersity is good, more namely removal core obtain the good hollow gold nanometer ball of monodispersity further.There is with the SERS probe that the hollow gold nanometer ball that this monodispersity is good is prepared from the immunoassay characteristic of very good homogeneity, sensitivity and repeatability.
In order to well solve the 3rd technical matters, we have invented the very high immunoassay of a kind of feasibility for repeatably detecting Tumor biomarkers fast as carcinomebryonic antigen (CEA) and alpha-fetoprotein (AFP) etc., the method is the hollow metal nanosphere modified based on SERS and by monoclonal antibody, this hollow metal nanosphere is formed by the core of the corroding metal nanoshell such as hydrofluorite, and it has showed the SERS activity stronger than gold nanoshell is small structure due to hollow metal nanostructured surface.Because whole course of reaction is all in the liquid phase, overcome the slow problem of reaction that the diffusion restriction brought at solid phase surface causes.Because its surperficial focus causes localized electromagnetic field to strengthen, hollow metal nanosphere serves good SERS and strengthens effect.Therefore, it is used to build SERS probe and for the high-sensitivity immunity analysis of biomarker, and the hollow gold nanometer ball of previously preparation is all be template by cobalt nanosphere usually, and at its superficial growth gold shell.But this method can not obtain the hollow metal nanosphere of homogeneity, has impact to quantitative test afterwards.Here, we propose a kind of new hollow metal nanosphere preparation method corrodes the good monodisperse gold shell (GNSs) of single dispersing silica core by hydrofluorite.With the SERS probe that the hollow metal nanosphere that this monodispersity is good is prepared from, there is very good homogeneity, sensitivity and repeatable and for immunoassay.CEA and AFP is one of a kind of modal tumor markers, and change of serum C EA and AFP also as the useful predicted data of preoperative test, can there are some researches show that tumor patient is higher than CEA and the AFP level of healthy population.
Technical scheme: in order to solve the problems of the technologies described above, technical scheme of the present invention is as follows: a kind of monodispersity hollow metal nanosphere, described monodispersity hollow metal nanosphere has the hollow kernel of monodispersity and the metal nano skin of monodispersity, described metal nano skin is one or more in gold nano layer, silver nanoparticle layer, copper nanometer layer or platinum nano-layers, and the hollow kernel of described monodispersity forms by corroding monodispersed organic or inorganic nanosphere.
Wherein, the hollow interior nuclear diameter of above-mentioned monodispersity is 40-800 nanometer.
Wherein, above-mentioned metal nano outer layer thickness is 20-500 nanometer.
The preparation method of above-mentioned a kind of monodispersity hollow metal nanosphere, comprises the following steps:
1) NaOH of 1 ~ 25 ml fresh configuration 1M adds in 500 ~ 1000 ml water, stir, and then 50-1000 μ l THPC is added, mixed liquor is obtained after magnetic agitation 2-20 minute, 5-100 ml 1% gold chloride adds in mixed liquor, namely magnetic agitation obtains the gold nano grain of 2 ~ 3 nm for 10 ~ 50 minutes, and 4 DEG C keep in Dark Place;
2) is stirred by 60 ~ 80 DEG C by the 3-aminopropyl triethoxysilane of the alcohol suspension of monodispersed organic or inorganic nanosphere and 100 ~ 400 μ l and carry out amination in 1 ~ 4 hour, ethanol washing is removed the 3-aminopropyl triethoxysilane that dissociates and is namely obtained amination nanosphere;
3) by through step 2) the amination nanosphere that processes adds the gold nano grain of step 1) process, and namely the magnetic agitation gold nano grain that after 10 ~ 50 minutes, centrifugal segregation does not adsorb obtains metal nano shell precursor;
4) be added in the salt solusion of gold, silver, copper or platinum by the metal nano shell precursor through step 3) process, adding reductive agent magnetic agitation after 10 ~ 50 minutes, namely centrifugal collecting precipitation obtains metal nano shell;
5) the metal nano shell through step 4) process is added in hydrofluorite or toluene removes kernel and namely obtain monodispersity hollow metal nanosphere.
Described organic or inorganic nanosphere is silica nanosphere, titanium dioxide nano-sphere, polystyrene nanospheres, polytetrafluorethylenano nano ball.
Further, the reductive agent in described step 4) is formaldehyde, hydrogen peroxide or hydroxylammonium chloride.
The application of above-mentioned a kind of monodispersity hollow metal nanosphere in preparation SERS probe.
Further, described SERS probe is Raman signal molecule by being adsorbed on monodispersity hollow metal nanosphere surface and conjugated monoclonal antibodies obtains.
Further, described monoclonal antibody is biomarker antibody.
Further, described biomarker antibody is Cea Monoclonal Antibodies or alpha-fetoprotein monoclonal antibody.Beneficial effect: compared with prior art, tool of the present invention has the following advantages:
(1) the present invention is compared with the hollow metal nanosphere invented in the past, and the SERS probe be prepared from based on monodispersity hollow metal nanosphere of the present invention has the immunoassay characteristic of very good homogeneity, sensitivity and repeatability.
(2) compared with existing hollow metal nanosphere, in hollow metal nanosphere preparation of the present invention, be commercially produced product as the inorganic of kernel or organic nano ball, various particle diameter has, and that is the hollow kernel of hollow metal nanosphere is adjustable; Also the thickness of metal level can be controlled in the preparation of metal nano shell.The plasmon resonance peak adjustable extent of the hollow metal nanosphere of such preparation is large, is conducive to obtaining larger SERS signal.
Accompanying drawing explanation
The extinction spectra of the hollow copper nanosphere of Fig. 1 (A) monodispersity and scanning electron microscopic picture; (B) the particle diameter statistic histogram of the hollow copper nanosphere of monodispersity;
Fig. 2 (A) adds the extinction spectra (after adding ion, the feature delustring peak of gold nanoshell disappears, and illustrates that gold nanoshell is assembled) of gold nanoshell before and after ion; (B) extinction spectra (after adding ion, the feature delustring peak of hollow gold nanometer ball still exists, and illustrates and does not assemble) of the hollow gold nanometer ball that mercaptoethanol is modified before and after ion is added;
The infared spectrum (showing that 4-mercaptobenzoic acid and monoclonal antibody are successfully modified on hollow gold nanometer ball) of the hollow gold nanometer ball of the hollow gold nanometer ball that Fig. 3 4-mercaptobenzoic acid is modified and monoclonal antibody coupling;
The scanning electron microscope (SEM) photograph of Fig. 4 (A) platinum nanoshell and schematic diagram; (B) scanning electron microscope (SEM) photograph of hollow platinum nanosphere and schematic diagram; (C) extinction spectra of platinum nanoshell and hollow platinum nanosphere; (D) the SERS spectrogram (show the enhancing effect of hollow platinum nanosphere than platinum nanoshell good) of platinum nanoshell and hollow platinum nanosphere;
The monodispersity hollow silver nanosphere SERS probe that Fig. 5 (A) CEA monoclonal antibody is modified is to the SERS spectrogram of the detection of CEA; (B) the monodispersity hollow copper nanosphere SERS probe of AFP monoclonal antibody modification is to the SERS spectrogram of the detection of AFP.
Embodiment
Embodiment 1:
A kind of monodispersity hollow metal nanosphere, monodispersity hollow metal nanosphere has the hollow kernel of monodispersity and the metal nano skin of monodispersity, described metal nano skin is gold nano layer, and the hollow kernel of monodispersity is formed by corrosion by titanium dioxide nano-sphere.The hollow interior nuclear diameter of monodispersity is 40 nanometers.Gold nano layer thickness is 20 nanometers.
A preparation method for monodispersity hollow metal nanosphere, comprises the following steps:
1) NaOH of 1ml fresh configuration 1 M adds in 500 ml water, stir, and then add 50 μ l THPCs, magnetic agitation after 2 minutes 5 ml 1% gold chlorides add in mixed liquor, namely magnetic agitation obtains the gold nano grain of 2 nm for 10 minutes, and 4 DEG C keep in Dark Place;
2) the 3-aminopropyl triethoxysilane of monodispersed titanium dioxide nano-sphere alcohol suspension and 100 μ l is stirred 4 h by 60 DEG C and carry out amination, ethanol washing is removed free 3-aminopropyl triethoxysilane and is namely obtained amination nanosphere.
3) by through step 2) the amination nanosphere that processes adds the gold nano grain of step 1) process, and namely the magnetic agitation golden nanometer particle that after 10 minutes, centrifugal segregation does not adsorb obtains metal nano shell precursor;
4) be added to by the metal nano shell precursor through step 3) process in the salt solusion of gold, adding formaldehyde as reductive agent magnetic agitation after 10 minutes, namely centrifugal collecting precipitation obtains metal nano shell;
5) gold nanoshell through step 4) process is added in hydrofluorite removes kernel and namely obtain monodispersity hollow gold nanometer ball.
Embodiment 2
A kind of monodispersity hollow metal nanosphere, monodispersity hollow metal nanosphere has the hollow kernel of monodispersity and the metal nano skin of monodispersity, described metal nano skin is silver nanoparticle layer, and the hollow kernel of described monodispersity is formed by corrosion by monodispersed polystyrene nanospheres.The hollow interior nuclear diameter of monodispersity is 800 nanometers.Silver nanoparticle layer thickness is 500 nanometers.
A preparation method for monodispersity hollow metal nanosphere, comprises the following steps:
1) NaOH of 25 ml fresh configuration 1 M adds in 1000 ml water, stir, and then add 1000 μ l THPCs, magnetic agitation after 20 minutes 100 ml 1% gold chlorides add in mixed liquor, namely magnetic agitation obtains the gold nano grain of 3 nm for 50 minutes, and 4 DEG C keep in Dark Place;
2) stirred by 80 DEG C by the 3-aminopropyl triethoxysilane of the alcohol suspension of monodispersed polystyrene nanospheres and 5000 μ l and carry out amination in 1 hour, ethanol washing is removed free 3-aminopropyl triethoxysilane and is namely obtained amination nanosphere.
3) by through step 2) the amination nanosphere that processes adds the gold nano grain of step 1) process, and namely the magnetic agitation golden nanometer particle that after 50 minutes, centrifugal segregation does not adsorb obtains metal nano shell precursor;
4) be added to by the metal nano shell precursor through step 3) process in the salt solusion of silver, adding hydrogen peroxide as reductive agent magnetic agitation after 50 minutes, namely centrifugal collecting precipitation obtains silver nanoparticle shell;
5) the silver nanoparticle shell through step 4) process is added in toluene removes kernel and namely obtain monodispersity hollow silver nanosphere.
Embodiment 3
A kind of monodispersity hollow metal nanosphere, monodispersity hollow metal nanosphere has the hollow kernel of monodispersity and the metal nano skin of monodispersity, described metal nano skin is copper nanometer layer, and the hollow kernel of described monodispersity is formed by corrosion by monodispersed silica nanosphere.The hollow interior nuclear diameter of monodispersity is 400 nanometers.Copper nanometer layer thickness is 250 nanometers.
A preparation method for monodispersity hollow metal nanosphere, comprises the following steps:
1) NaOH of 12 ml fresh configuration 1 M adds in 720 ml water, stir, and then add 500 μ l THPCs, magnetic agitation after 10 minutes 50 ml 1% gold chlorides add in mixed liquor, namely magnetic agitation obtains the gold nano grain of 2 nm for 30 minutes, and 4 DEG C keep in Dark Place;
2) stirred by 70 DEG C by the 3-aminopropyl triethoxysilane of the alcohol suspension of monodispersed silica nanosphere and 2500 μ l and carry out amination in 3 hours, ethanol washing is removed free 3-aminopropyl triethoxysilane and is namely obtained amination nanosphere.
3) by through step 2) the amination nanosphere that processes adds the gold nano grain of step 1) process, and namely the magnetic agitation golden nanometer particle that after 30 minutes, centrifugal segregation does not adsorb obtains metal nano shell precursor;
4) be added in the salt solusion of copper by the metal nano shell precursor through step 3) process, adding hydroxylammonium chloride as reductive agent magnetic agitation after 30 minutes, namely centrifugal collecting precipitation obtains copper nanoshell;
5) the copper nanoshell through step 4) process is added in hydrofluorite removes kernel and namely obtain monodispersity hollow copper nanosphere (see figure 1).
Embodiment 4
In the same manner as in Example 1, difference is, step 2) in 3-aminopropyl triethoxysilane amount be 400 μ l, the salt solusion of what step 4) added is platinum, what obtain is the hollow platinum nanosphere of monodispersity, and described monodispersed hollow kernel is formed by corrosion by monodispersed polytetrafluorethylenano nano ball.Fig. 4 shows that the enhancing effect of hollow platinum nanosphere is better than platinum nanoshell.
The monodispersity hollow metal nanosphere preparation that embodiment 5 SERS probe--carcinomebryonic antigen (CEA) monoclonal antibody is modified
We select 4-mercaptobenzoic acid as Raman signal molecule, are also the coupling agents connecting CEA monoclonal antibody simultaneously.The 4-mercaptobenzoic acid (10 that general 10 μ l newly configure
-3m) join the monodispersity hollow metal nanosphere of 7 ml, stir after 30 minutes, Raman signal molecule is adsorbed on hollow gold goal surface by the effect of sulfydryl and Au.We cover other non-specific sites with the mercaptoethanol (2.5 mM newly prepare) of 2 μ l, can prevent these sites and other albumen effects like this, this improves the stability (as shown in Figure 2) of hollow gold nanometer ball simultaneously; The mercaptoethanol that centrifugal segregation is unnecessary, PBS resuspension is used after washing three times with PBS damping fluid, the water-soluble carbodiimide (EDC) of last 5 μ l and nitrogen hydroxysuccinimide (NHS) add solution to activate the carboxyl of 4-mercaptobenzoic acid end, after 30 minutes, 50 μ lCEA monoclonal antibodies (40 μ g/ml) add the hollow gold goal of NHS-activation, react 2 hours.Carcinomebryonic antigen (CEA) monoclonal antibody is fixed on the surface of hollow metal nanosphere by the carboxyl of activation.
The sign of the monodispersity hollow silver nanosphere SERS probe that embodiment 6 CEA monoclonal antibody is modified
The monodispersity hollow metal nanosphere that CEA monoclonal antibody is modified carries out IR Characterization, and Fig. 3 provides monodispersity hollow silver nanosphere infrared spectrum (redness) of 4-mercaptobenzoic acid modification and monodispersity hollow silver nanosphere infrared spectrum (black) of monoclonal antibody modification.At 3729 cm in spectrum
-1with 675 cm
-1place appears at the flexural vibrations that two spectral lines are hydroxyls, shows that mercaptoethanol is successfully modified on monodispersity hollow metal nanosphere.At 1600 cm
-1the peak value at place appears at the phenyl ring skeletal vibration that two spectral lines correspond to 4-mercaptobenzoic acid simultaneously, but 1126 cm
-1with 1374 cm
-1spectral line be the C-N of primary amine and the resonance peak of acid amides C-N, instruction book clonal antibody has been coupled on the 4-mercaptobenzoic acid on monodispersity hollow silver nanosphere.
The monodispersity hollow silver nanosphere SERS probe that embodiment 7 carcinomebryonic antigen (CEA) monoclonal antibody is modified is applied the detection of CEA
The magnetic microsphere of the antibody coupling of 0.5 mg/ml of first, 10 μ L is stirring 20 minutes containing in CEA antigen 10 μ L solution to be measured; The second, magnetic resolution catches the magnetic microsphere of CEA, with PBS buffer solution three times; 3rd, the monodispersity hollow silver nanosphere SERS probe (OD that the magnetic microsphere obtained is modified with 10 μ LCEA monoclonal antibodies further
700nm=1) stir 20 minutes under.Finally, in magnetic resolution and be dispersed in after washing three times 10 μ L PBS buffer solution formed sandwich immunoassay compound suspending liquid.Do not have CEA as blank group, BSA, as negative control group, is shown in Fig. 5 (A), the SERS signal of acquisition with treat that the CEA concentration in test sample is linear.
The hollow platinum nanosphere preparation of monodispersity that embodiment 8 SERS probe-alpha-fetoprotein (AFP) monoclonal antibody is modified
We select 4-mercaptobenzoic acid as Raman signal molecule, are also the coupling agents connecting AFP monoclonal antibody simultaneously.The 4-mercaptobenzoic acid (10 that general 10 μ l newly configure
-3m) join the hollow platinum nanosphere of 7 ml, stir after 30 minutes, Raman signal molecule is adsorbed on hollow platinum nanosphere surface by the effect of sulfydryl and Au.We cover other non-specific sites with the mercaptoethanol (2.5 mM newly configure) of 2 μ l, can prevent these sites and other albumen effects like this, this improves the stability of hollow platinum nanosphere simultaneously; The mercaptoethanol that centrifugal segregation is unnecessary, PBS resuspension is used after washing three times with PBS damping fluid, the water-soluble carbodiimide (EDC) of last 5 μ l and nitrogen hydroxysuccinimide (NHS) add solution to activate the carboxyl of 4-mercaptobenzoic acid end, after 30 minutes, 50 μ l AFP monoclonal antibodies (40 μ g/ml) add the hollow platinum nanosphere of NHS-activation, react 2 hours.Alpha-fetoprotein (AFP) monoclonal antibody is fixed on the surface of hollow platinum nanosphere by the carboxyl of activation.
The hollow platinum nanosphere of monodispersity that embodiment 9 alpha-fetoprotein (AFP) monoclonal antibody is modified detects application
The magnetic microsphere of the antibody coupling of 0.5 mg/ml of first, 10 μ L is stirring 20 minutes containing in AFP antigen 10 μ L solution to be measured; The second, magnetic resolution catches the magnetic microsphere of AFP, with PBS buffer solution three times; 3rd, the monodispersity hollow platinum nanosphere SERS probe (OD that the magnetic microsphere obtained is modified with 10 μ LAFP monoclonal antibodies further
700nm=1) stir 20 minutes under.Finally, in magnetic resolution and be dispersed in after washing three times 10 μ L PBS buffer solution formed sandwich immunoassay compound suspending liquid.Do not have AFP as blank group, BSA, as negative control group, is shown in Fig. 5 (B), the SERS signal of acquisition with treat that the AFP concentration in test sample is linear.
Following non-limiting example can make the present invention of those of ordinary skill in the art's comprehend, but does not limit the present invention in any way.
Claims (10)
1. a monodispersity hollow metal nanosphere, it is characterized in that, described monodispersity hollow metal nanosphere has the hollow kernel of monodispersity and the metal nano skin of monodispersity, described metal nano skin is one or more in gold nano layer, silver nanoparticle layer, copper nanometer layer or platinum nano-layers, and the hollow kernel of described monodispersity forms by corroding monodispersed organic or inorganic nanosphere.
2. a kind of monodispersity hollow metal nanosphere according to claim 1, is characterized in that, the hollow interior nuclear diameter of described monodispersity is 40-800 nanometer.
3. a kind of monodispersity hollow metal nanosphere according to claim 1, it is characterized in that, described metal nano outer layer thickness is 20-500 nanometer.
4. the preparation method of a kind of monodispersity hollow metal nanosphere according to claim 1, is characterized in that, comprise the following steps:
1) NaOH of 1 ~ 25 ml fresh configuration 1M adds in 500 ~ 1000 ml water, stir, and then 50-1000 μ l THPC is added, mixed liquor is obtained after magnetic agitation 2-20 minute, 5-100 ml 1% gold chloride adds in mixed liquor, namely magnetic agitation obtains the gold nano grain of 2 ~ 3 nm for 10 ~ 50 minutes, and 4 DEG C keep in Dark Place;
2) is stirred by 60 ~ 80 DEG C by the 3-aminopropyl triethoxysilane of the alcohol suspension of monodispersed organic or inorganic nanosphere and 100 ~ 5000 μ l and carry out amination in 1 ~ 4 hour, ethanol washing is removed the 3-aminopropyl triethoxysilane that dissociates and is namely obtained amination nanosphere;
3) by through step 2) the amination nanosphere that processes adds the gold nano grain of step 1) process, and namely the magnetic agitation gold nano grain that after 10 ~ 50 minutes, centrifugal segregation does not adsorb obtains metal nano shell precursor;
4) be added in the salt solusion of gold, silver, copper or platinum by the metal nano shell precursor through step 3) process, adding reductive agent magnetic agitation after 10 ~ 50 minutes, namely centrifugal collecting precipitation obtains metal nano shell;
5) the metal nano shell through step 4) process is added in hydrofluorite or toluene removes kernel and namely obtain monodispersity hollow metal nanosphere.
5. the preparation method of a kind of monodispersity hollow metal nanosphere according to claim 4, it is characterized in that, described organic or inorganic nanosphere is silica nanosphere, titanium dioxide nano-sphere, polystyrene nanospheres, polytetrafluorethylenano nano ball.
6. the preparation method of a kind of monodispersity hollow metal nanosphere according to claim 4, it is characterized in that, the reductive agent in described step 4) is formaldehyde, hydrogen peroxide or hydroxylammonium chloride.
7. the application of a kind of monodispersity hollow metal nanosphere according to claim 1 in preparation SERS probe.
8. application according to claim 7, is characterized in that, described SERS probe is Raman signal molecule by being adsorbed on monodispersity hollow metal nanosphere surface and conjugated monoclonal antibodies obtains.
9. application according to claim 7, is characterized in that, described monoclonal antibody is biomarker antibody.
10. application according to claim 7, is characterized in that, described biomarker antibody is Cea Monoclonal Antibodies or alpha-fetoprotein monoclonal antibody.
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| CN108356279A (en) * | 2018-03-09 | 2018-08-03 | 华中科技大学 | A kind of preparation method of hollow gold nanometer material |
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| CN115415539A (en) * | 2022-08-11 | 2022-12-02 | 华测检测认证集团股份有限公司 | Preparation and application methods of hollow ruthenium-copper alloy nanoparticles |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8075664B1 (en) * | 2007-06-11 | 2011-12-13 | Sandia Corporation | Synthesis of metallic nanoshells on porphyrin-stabilized emulsions |
| US20130230570A1 (en) * | 2007-08-14 | 2013-09-05 | The Regents Of The University Of California | Hollow silica nanospheres and methods of making same |
-
2014
- 2014-10-24 CN CN201410577586.8A patent/CN104360064A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8075664B1 (en) * | 2007-06-11 | 2011-12-13 | Sandia Corporation | Synthesis of metallic nanoshells on porphyrin-stabilized emulsions |
| US8568508B1 (en) * | 2007-06-11 | 2013-10-29 | Sandia Corporation | Metallic nanoshells on porphyrin-stabilized emulsions |
| US20130230570A1 (en) * | 2007-08-14 | 2013-09-05 | The Regents Of The University Of California | Hollow silica nanospheres and methods of making same |
Non-Patent Citations (7)
Cited By (10)
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| CN105482803A (en) * | 2015-11-26 | 2016-04-13 | 东南大学 | Fluorescence-SERS dual-mode super-resolution imaging probe and its preparation method and use method |
| CN108414758A (en) * | 2018-01-12 | 2018-08-17 | 宁波大学 | Preparation method and applications for the SERS biosensors for detecting tumor markers miRNA-141 |
| CN108414758B (en) * | 2018-01-12 | 2020-06-19 | 宁波大学 | Preparation method and application of SERS biosensor for detecting tumor marker miRNA-141 |
| CN108344713A (en) * | 2018-02-06 | 2018-07-31 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Photonic crystal sensor material and its preparation method and application |
| CN108344713B (en) * | 2018-02-06 | 2021-04-13 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Photonic crystal sensing material and preparation method and application thereof |
| CN108356279A (en) * | 2018-03-09 | 2018-08-03 | 华中科技大学 | A kind of preparation method of hollow gold nanometer material |
| CN112296350A (en) * | 2020-09-27 | 2021-02-02 | 浙江师范大学 | Magnetic hollow microsphere and preparation method and application thereof |
| CN114264805A (en) * | 2021-12-06 | 2022-04-01 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of SERS enhancement-based probe, product and application thereof |
| CN115415539A (en) * | 2022-08-11 | 2022-12-02 | 华测检测认证集团股份有限公司 | Preparation and application methods of hollow ruthenium-copper alloy nanoparticles |
| CN116275016A (en) * | 2022-09-07 | 2023-06-23 | 深圳达闼科技控股有限公司 | Method for detecting hydrogen peroxide content and pH value by nano particles and surface enhanced Raman scattering |
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