CN102672167B - Preparation and application of novel triangular sliver-silver sulfide nanocomposite particle - Google Patents
Preparation and application of novel triangular sliver-silver sulfide nanocomposite particle Download PDFInfo
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- CN102672167B CN102672167B CN201110063379.7A CN201110063379A CN102672167B CN 102672167 B CN102672167 B CN 102672167B CN 201110063379 A CN201110063379 A CN 201110063379A CN 102672167 B CN102672167 B CN 102672167B
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Abstract
The invention provides a preparation method of a novel triangular sliver-silver sulfide nanocomposite particle and application of the novel triangular sliver-silver sulfide nanocomposite particle in oligonucleotide probe synthesis. The novel triangular sliver-silver sulfide nanocomposite particle is prepared by a triangular silver nanoparticle and sulfide through direct reaction, and a protective layer formed by silver sulfide is covered on the surface of the triangular silver nanoparticle. The invention also provides a preparation method of the nanocomposite particle, which is characterized in that the process is simple and quick; due to the synthesized nanoparticle, not only can the stability of the triangular sliver nanoparticle be maintained, but also the sensitivity of chemical and biometric identification is not influenced, the corresponding biological nano probe can be prepared by coupling the silver sulfide on the surface of the triangular sliver-silver sulfide nanocomposite particle and the oligonucleotide probe modified by sulfydryl without carrying out surface modification on the nanoparticle, the process is simple and quick, and the preparation method has wide application prospect in the biological sensing field.
Description
Technical field
The present invention relates to a kind of preparation method of noble metal nano composite particles and the application in preparing oligonucleotide probe thereof, relate in particular to a kind of preparation method of Triangular Silver-silver sulfide Nano composite granules and the application in preparing oligonucleotide probe thereof.
Background technology
Noble metal nano particles (as gold and silver nano-grain) has potential wide application prospect in catalysis, optics and bio-sensing.Metal is received to research, the especially research to the controlled preparation of its pattern and corresponding properties and application of particle, is the forward position focus of material science and association area always.Because Triangular Silver nano particle has, unique surperficial proton resonance (SPR) character-it has three obvious SPR peaks, i.e. bipolar formant in four utmost points and face in outer four utmost points of face, face, and this character and its size and dimension are closely related.Therefore, for the research of triangle silver nano-grain, attracted increasing people's notice.One of them important application is that the direct function of Triangular Silver nano particle and biomolecule obtains corresponding biological nano probe.
Yet Triangular Silver nano particle has very high surface energy, particularly in wedge angle and edge, Ag atom is herein as easy as rolling off a log oxidized.This oxidation can cause subsiding of wedge angle in its structure or the integral body of structure to be dissolved, and SPR moves at peak even and disappears.This makes Triangular Silver nano particle may directly apply to biological field hardly.In order to realize it, further apply, need to improve the stability of varying environment intermediate cam shape silver nano-grain.Although the passivation layer of mercaptan and silica can and improve their stability for the protection of Triangular Silver nano particle, these methods have following shortcoming: 1) process complexity is loaded down with trivial details; 2) group of passivation can affect the SPR sensitivity of stability and the detection molecules of biomolecule.
Therefore, need to find a kind ofly can keep Triangular Silver nano particle stability, not affect again the method for its chemistry and bio-identification sensitivity.
Summary of the invention
In order to solve the defect of above-mentioned prior art, the object of the present invention is to provide and a kind ofly take Triangular Silver nano particle and prepare triangle Ag-Ag as raw material
2the preparation method of S Nano composite granules and oligonucleotide probe thereof.Feature of the present invention is simple and quick, and has good reappearance.And further by triangle Ag-Ag
2the Ag on S Nano composite granules surface
2corresponding oligonucleotide probe is prepared in the oligonucleotides of S and sulfydryl modification (HS-DNA) coupling.
The present invention adopts following technical scheme in order to achieve the above object:
The invention provides a kind of New triangular silver-silver sulfide Nano composite granules; wherein comprise Triangular Silver nano particle; it is characterized in that: it is directly to react preparation by Triangular Silver nano particle with sulfide in the surface coverage of Triangular Silver nano particle, having the protective layer of silver sulfide formation.
Described silver sulfide can directly be prepared corresponding oligonucleotide probe with the oligonucleotides phase coupling of sulfydryl modification.
Its preparation method comprises the following steps:
(1) prepare the Triangular Silver nano particle of polyvinylpyrrolidone (PVP) protection;
(2) under lucifuge condition, in the solution of step (1), add fast the aqueous solution of sulfide, constant temperature keeps 20-35 ℃, stirs 5-30 minute.
Wherein step (1) can adopt existing method, Adv Mater for example, 2005,17 (4): the disclosed method of 412-415 prepares.
Wherein in step (2), add the aqueous solution of sulfide, the mol ratio that makes Triangular Silver nano particle and sulfide is 50: 1-5: 1.
Wherein in step (2), sulfide used is vulcanized sodium or potassium sulfide.
In addition, the present invention also provides the application process of this New triangular silver-silver sulfide Nano composite granules in preparing oligonucleotide probe, comprises the following steps:
(1) triangle Ag-Ag
2s Nano composite granules washs with phosphate buffer;
(2) under room temperature, by triangle Ag-Ag
2s Nano composite granules mixes with oligonucleotides (HS-DNA) and the borate buffer of sulfydryl modification, the centrifugal supernatant of removing;
(3) phosphate buffer washing for precipitation, obtains triangle Ag-Ag
2s Nano composite granules oligonucleotide probe;
(4) the triangle Ag-Ag obtaining
2s Nano composite granules oligonucleotide probe is heavily distributed in phosphate buffer, in 4 ℃ of storages.
Wherein, the pH value of step (1) phosphate buffer used is 7.5-8.5, contains the NaCl that concentration is 0.1-0.2mol/L; Preferably pH value is 8.0, contains the NaCl that concentration is 0.15mol/L.
Wherein, in step (3) and (4), the pH value of phosphate buffer used is 6.5-7.5, contains the NaCl that concentration is 0.1-0.2mol/L; Preferably pH value is 7.0, contains the NaCl that concentration is 0.15mol/L.
Wherein, in step (2), the pH value of borate buffer used is 8.5-9.5, and concentration is 45-55mmol/L; Preferably pH value is 9.2, and concentration is 50mmol/L.
The invention has the advantages that:
(1) provide a kind of triangle Ag-Ag of simple and quick synthesizing new
2the preparation method of S Nano composite granules;
(2) by triangle Ag-Ag
2the Ag on S Nano composite granules surface
2corresponding oligonucleotide probe is prepared in S and HS-DNA coupling.
Utilize the prepared triangle Ag-Ag of the present invention
2s Nano composite granules is prepared the process of oligonucleotide probe and has been saved the process that nano material itself is modified again, has outstanding advantage simply and easily.The New triangular Ag-Ag that the present invention is synthetic
2s Nano composite granules can be used for the detection of DNA molecular, and it is with a wide range of applications in bio-sensing field as novel nano bioprobe.
Accompanying drawing explanation
Fig. 1 is triangle Ag-Ag of the present invention
2s Nano composite granules oligonucleotide probe is prepared schematic diagram.
Fig. 2 is triangle Ag-Ag prepared by one embodiment of the invention
2the picture of the transmission electron microscope of S Nano composite granules (TEM).
Fig. 3 is triangle Ag-Ag prepared by one embodiment of the invention
2the melting temperature schematic diagram of S nano particle oligonucleotide probe and target DNA.
Fig. 4 is triangle Ag-Ag prepared by further embodiment of this invention
2the picture of the transmission electron microscope of S Nano composite granules (TEM).
Fig. 5 is triangle Ag-Ag prepared by further embodiment of this invention
2the melting temperature schematic diagram of S nano particle oligonucleotide probe and target DNA.
Fig. 6 is triangle Ag-Ag prepared by another embodiment of the present invention
2the picture of the transmission electron microscope of S Nano composite granules (TEM).
Fig. 7 is the triangle Ag-Ag2S nano particle oligonucleotide probe prepared of another embodiment of the present invention and the melting temperature schematic diagram of target DNA.
The specific embodiment
Embodiment 1:
Triangle Ag-Ag
2the preparation of S Nano composite granules:
(1) reference literature (Adv Mater, 2005,17 (4): method 412-415.) is prepared the triangle silver nano-grain of PVP protection: use PVP and citrate as stabilizing agent and masterplate agent, use NaBH under lucifuge stirring condition
4reduction AgNO
3prepare the triangle silver nano-grain of PVP protection.
(2) in the triangle silver nano-grain solution of preparing in step (1) under lucifuge condition, add fast 1 μ L1mmol/L sodium sulfide solution, the molar ratio of triangle silver nano-grain and vulcanized sodium is approximately 10: 1.25 ℃ of stirrings of constant temperature 30 minutes, obtain triangle Ag-Ag
2s Nano composite granules.Its pattern is shown in accompanying drawing 2.
Utilize triangle Ag-Ag
2s Nano composite granules is prepared oligonucleotide probe and cross performance test case thereof:
Model oligonucleotide molecules is as follows:
Probe a:5 '-TCT-CAA-CTC-GTA-TTTT-SH-3 '
Probe b:5 '-SH-TTTT-CGC-ATT-CAG-GAT-3 '
Target DNA:5 '-TAC-GAG-TTG-AGA-ATC-CTG-AAT-GCG-3 '
(1) the triangle Ag-Ag being prepared by Fig. 1
2s is phosphate buffer (PBS) (pH 8.0, NaCl 0.15mol/L) washing for Nano composite granules.
(2) under room temperature condition, by the triangle Ag-Ag of step (1)
2s Nano composite granules and probe a and borate buffer (50mmol/L, pH 9.2) mix, the centrifugal supernatant of removing, and precipitation is cleaned with phosphate buffer (pH 7.0, NaCl 0.15mol/L).By the oligonucleotides-modified triangle Ag-Ag of the sulfydryl obtaining
2(nano-probe a) is heavily distributed in phosphate buffer and in 4 ℃ of storages S nano particle.
(3) nano-probe b reacts preparation by same method with probe b, is heavily scattered in phosphate buffer and in 4 ℃ of storages.
(4) nano-probe a and b are mixed with target DNA, be heated to 70 ℃ above 10 minutes, naturally cooling, use spectrophotometer detects its melting temperature.See accompanying drawing 3.
Embodiment 2:
Triangle Ag-Ag
2the preparation of S Nano composite granules:
Preparation method and example 1 are basic identical; Difference is: in step (2) under lucifuge condition in step 1) add fast the sodium sulfide solution of 3 μ L 2mmol/L in the triangle silver nano-grain solution prepared, the molar ratio of triangle silver nano-grain and vulcanized sodium is approximately 20: 1.30 ℃ of stirrings of constant temperature 15 minutes, obtain triangle Ag-Ag after reaction
2s Nano composite granules.Its pattern is shown in accompanying drawing 4.
Utilize triangle Ag-Ag
2s Nano composite granules is prepared oligonucleotide probe and cross performance test case thereof:
Model oligonucleotide molecules is as follows:
Probe a:5 '-CAA-TCT-CTC-GTA-TTTT-SH-3 '
Probe b:5 '-SH-TTTT-AGT-CAG-ATT-GAT-3 '
Target DNA:5 '-TAC-GAG-AGA-TTG-ATC-AAT-CTG-ACT-3 '
(1) the triangle Ag-Ag being prepared by Fig. 4
2s is phosphate buffer (PBS) (pH 8.0, NaCl 0.15mol/L) washing for Nano composite granules.
(2) under room temperature condition, by the triangle Ag-Ag of step (1)
2s Nano composite granules and probe a and borate buffer (50mmol/L, pH 9.2) mix, the centrifugal supernatant of removing, and precipitation is cleaned with phosphate buffer (pH 7.0, NaCl 0.15mol/L).By the oligonucleotides-modified triangle Ag-Ag of the sulfydryl obtaining
2(nano-probe a) is heavily distributed in phosphate buffer and in 4 ℃ of storages S nano particle.
(3) nano-probe b reacts preparation by same method with probe b, is heavily scattered in phosphate buffer and in 4 ℃ of storages.
(4) nano-probe a and b are mixed with target DNA, be heated to 60 ℃ above 10 minutes, naturally cooling, use spectrophotometer detects its melting temperature.See accompanying drawing 5.
Embodiment 3:
Triangle Ag-Ag
2the preparation of S Nano composite granules:
Preparation method and example 1 are basic identical; Difference is: in step (2) under lucifuge condition in step 1) add fast the potassium sulfide aqueous solution of 2 μ L 3mmol/L in the triangle silver nano-grain solution prepared, the molar ratio of triangle silver nano-grain and potassium sulfide is approximately 30: 1.20 ℃ of stirrings of constant temperature 25 minutes, obtain triangle Ag-Ag after reaction
2s Nano composite granules.Its pattern is shown in accompanying drawing 6.
Utilize triangle Ag-Ag
2s Nano composite granules is prepared oligonucleotide probe and cross performance test case thereof:
Model oligonucleotide molecules is as follows:
Probe a:5 '-CAC-TCT-CTC-GTA-TTTT-SH-3 '
Probe b:5 '-SH-TTTT-AGT-CAG-ATT-GAG-3 '
Target DNA:5 '-TAC-GAG-AGA-GTG-CTC-AAT-CTG-ACT-3 '
(1) the triangle Ag-Ag being prepared by Fig. 6
2s is phosphate buffer (PBS) (pH 8.0, NaCl 0.15mol/L) washing for Nano composite granules.
(2) under room temperature condition, by the triangle Ag-Ag of step (1)
2s Nano composite granules and probe a and borate buffer (50mmol/L, pH 9.2) mix, the centrifugal supernatant of removing, and precipitation is cleaned with phosphate buffer (pH 7.0, NaCl 0.15mol/L).By the oligonucleotides-modified triangle Ag-Ag of the sulfydryl obtaining
2(nano-probe a) is heavily distributed in phosphate buffer and in 4 ℃ of storages S nano particle.
(3) nano-probe b reacts preparation by same method with probe b, is heavily scattered in phosphate buffer and in 4 ℃ of storages.
(4) nano-probe a and b are mixed with target DNA, be heated to 80 ℃ above 5 minutes, naturally cooling, use spectrophotometer detects its melting temperature.See accompanying drawing 7.
The triangle Ag-Ag that embodiment of the present invention 1-3 prepares
2s Nano composite granules oligonucleotide probe has preparation easy than the oligonucleotide probe that three horn silver AgXs are prepared as precursor of take of having reported, the feature easily repeating, and can obviously find out from the melting temperature curve of accompanying drawing 3,5,7: the sudden change by monitoring absorbance just can obtain melting temperature numerical value very accurately, and this illustrates that this probe will be a kind of very desirable material in following oligonucleotides context of detection.
Claims (12)
1. Triangular Silver-silver sulfide Nano composite granules; wherein comprise Triangular Silver nano particle; it is characterized in that: in the surface coverage of Triangular Silver nano particle, have the protective layer of silver sulfide formation, described silver sulfide directly reacts preparation by Triangular Silver nano particle with sulfide.
2. Triangular Silver-silver sulfide Nano composite granules as claimed in claim 1, is characterized in that described silver sulfide can directly prepare corresponding oligonucleotide probe with the oligonucleotides phase coupling of sulfydryl modification.
3. a method of preparing Triangular Silver-silver sulfide Nano composite granules as claimed in claim 1 or 2,, it is characterized in that mainly comprising the following steps:
(1) prepare the Triangular Silver nano particle of polyvinylpyrrolidone (PVP) protection;
(2) under lucifuge condition, in the solution of step (1), add fast the aqueous solution of sulfide, constant temperature keeps 20-35 ℃, stirs 5-30 minute.
4. preparation method as claimed in claim 3, is characterized in that: the aqueous solution of the sulfide adding in described step (2), the mol ratio that makes Triangular Silver nano particle and sulfide is 50:1-5:1.
5. preparation method as claimed in claim 3, is characterized in that: in described step (2), sulfide used is vulcanized sodium or potassium sulfide.
6. the application of the Triangular Silver-silver sulfide Nano composite granules described in claim 1-5 any one, is prepared corresponding oligonucleotide probe, comprises the following steps:
(1) triangle Ag-Ag
2s Nano composite granules washs with phosphate buffer;
(2) under room temperature, by triangle Ag-Ag
2s Nano composite granules mixes with oligonucleotides and the borate buffer of sulfydryl modification, the centrifugal supernatant of removing;
(3) phosphate buffer washing for precipitation, obtains triangle Ag-Ag
2s Nano composite granules oligonucleotide probe;
(4) the triangle Ag-Ag obtaining
2s Nano composite granules oligonucleotide probe is heavily distributed in phosphate buffer, in 4 ℃ of storages.
7. application as claimed in claim 6, is characterized in that, the pH value of step (1) phosphate buffer used is 7.5-8.5, contains the NaCl that concentration is 0.1-0.2mol/L.
8. application as claimed in claim 7, is characterized in that, the pH value of step (1) phosphate buffer used is 8.0, contains the NaCl that concentration is 0.15mol/L.
9. application as claimed in claim 6, is characterized in that, in step (3) and (4), the pH value of phosphate buffer used is 6.5-7.5, contains the NaCl that concentration is 0.1-0.2mol/L.
10. application as claimed in claim 9, is characterized in that, in step (3) and (4), the pH value of phosphate buffer used is 7.0, contains the NaCl that concentration is 0.15mol/L.
11. application as claimed in claim 6, is characterized in that, in step (2), the pH value of borate buffer used is 8.5-9.5, and concentration is 45-55mmol/L.
12. application as claimed in claim 11, is characterized in that, in step (2), the pH value of borate buffer used is 9.2, and concentration is 50mmol/L.
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| CN103785848B (en) * | 2013-12-26 | 2016-08-24 | 江苏大学 | A kind of method of one-pot synthesis Ag-Ag2S/CdS heterojunction structure and the application of Ag-Ag2S/CdS heterojunction structure |
| CN105412940B (en) * | 2015-12-02 | 2018-10-12 | 鲁东大学 | A kind of composite nano antibacterial material is used for the treatment of drug resistance of vancomycin pathogenic bacteria |
| CN107159881B (en) * | 2017-06-12 | 2020-04-07 | 青岛科技大学 | Silver @ silver sulfide core-shell structure photothermal conversion nanomaterial |
| CN108587599A (en) * | 2018-03-28 | 2018-09-28 | 苏州星烁纳米科技有限公司 | Quantum dot dispersion |
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| US7144627B2 (en) * | 1997-03-12 | 2006-12-05 | William Marsh Rice University | Multi-layer nanoshells comprising a metallic or conducting shell |
| CN101165469A (en) * | 2006-10-19 | 2008-04-23 | 河南师范大学 | Protein coated silver sulfide nano line preparation method |
| CN101274751A (en) * | 2007-03-30 | 2008-10-01 | 清华大学 | Preparation for monodisperse, argentic sulfide and silver selenide nanocrystal |
| CN101525669A (en) * | 2009-03-11 | 2009-09-09 | 中国人民解放军第三军医大学第一附属医院 | Nuclear acid probe marked with quantum-dots composite sphere and preparation method and application thereof |
| CN101947655A (en) * | 2010-10-25 | 2011-01-19 | 江苏技术师范学院 | Preparation method of triangular silver nanosheet |
| CN102009172A (en) * | 2010-09-27 | 2011-04-13 | 山东大学 | Resistance switch material in Ag/Ag2S core/shell nanometer structure and preparation method thereof |
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| WO2002096262A2 (en) * | 2001-05-25 | 2002-12-05 | Northwestern University | Non-alloying core shell nanoparticles |
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7144627B2 (en) * | 1997-03-12 | 2006-12-05 | William Marsh Rice University | Multi-layer nanoshells comprising a metallic or conducting shell |
| CN101165469A (en) * | 2006-10-19 | 2008-04-23 | 河南师范大学 | Protein coated silver sulfide nano line preparation method |
| CN101274751A (en) * | 2007-03-30 | 2008-10-01 | 清华大学 | Preparation for monodisperse, argentic sulfide and silver selenide nanocrystal |
| CN101525669A (en) * | 2009-03-11 | 2009-09-09 | 中国人民解放军第三军医大学第一附属医院 | Nuclear acid probe marked with quantum-dots composite sphere and preparation method and application thereof |
| CN102009172A (en) * | 2010-09-27 | 2011-04-13 | 山东大学 | Resistance switch material in Ag/Ag2S core/shell nanometer structure and preparation method thereof |
| CN101947655A (en) * | 2010-10-25 | 2011-01-19 | 江苏技术师范学院 | Preparation method of triangular silver nanosheet |
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