CN102672167A - 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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- CN102672167A CN102672167A CN2011100633797A CN201110063379A CN102672167A CN 102672167 A CN102672167 A CN 102672167A CN 2011100633797 A CN2011100633797 A CN 2011100633797A CN 201110063379 A CN201110063379 A CN 201110063379A CN 102672167 A CN102672167 A CN 102672167A
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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 and the application in the preparation oligonucleotide probe thereof of noble metal nano composite particles, relate in particular to the preparation method and the application in the preparation oligonucleotide probe thereof of a kind of triangle silver-silver sulfide Nano composite granules.
Background technology
Noble metal nano particles (like gold and silver nano-grain) has potential wide application prospect in catalysis, optics and bio-sensing.Metal is received the research of particle, especially to the research of the controlled preparation of its pattern and corresponding properties and application, is the forward position focus of material science and association area always.Unique surperficial proton resonance (SPR) character-it has three tangible SPR peaks because of the triangle silver nano-grain has, i.e. bipolar formant in four utmost points and the face in outer four utmost points of face, the face, and this character and its size and dimension are closely related.Therefore, attracted increasing people's notice for the research of triangle silver nano-grain.One of them important use is that the direct functionization of triangle silver nano-grain and biomolecule obtains corresponding biological nano probe.
Yet the triangle silver nano-grain has very high surface energy, and particularly in wedge angle and edge, the Ag atom here is as easy as rolling off a log oxidized.This oxidation can cause subsiding of wedge angle in its structure or the dissolving of the integral body of structure, and SPR moves at the peak even disappears.This makes the triangle silver nano-grain possibly directly apply to biological field hardly.In order to realize its further application, need to improve the stability of varying environment intermediate cam shape silver nano-grain.Although the passivation layer of mercaptan and silica can be used for the protection of triangle silver nano-grain and improve their stability, these methods have following shortcoming: 1) the process complicacy is loaded down with trivial details; 2) group of passivation can influence the SPR sensitivity of the stability and the detection molecules of biomolecule.
Therefore, need find a kind of triangle silver nano-grain that can keep stable, not influence its chemistry and bio-identification sensitivity of method again.
Summary of the invention
In order to solve the defective of above-mentioned prior art, the object of the present invention is to provide a kind of is feedstock production triangle Ag-Ag with the triangle silver nano-grain
2The preparation method of S Nano composite granules and oligonucleotide probe thereof.Characteristics of the present invention are simple and quick, and have good reappearance.And further through 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 present invention provides a kind of New triangular silver-silver sulfide Nano composite granules; Wherein comprise the triangle silver nano-grain; It is characterized in that: it is by triangle silver nano-grain and the direct prepared in reaction of sulfide, and the protective layer of silver sulfide formation is arranged in the surface coverage of triangle silver nano-grain.
Described silver sulfide can be directly with the oligonucleotides of sulfydryl modification mutually coupling prepare corresponding oligonucleotide probe.
Its preparation method may further comprise the steps:
(1) the triangle silver nano-grain of preparation polyvinylpyrrolidone (PVP) protection;
(2) in the solution of step (1), add fast the aqueous solution of sulfide under the lucifuge condition, constant temperature keeps 20-35 ℃, stirs 5-30 minute.
Wherein step (1) can adopt existing method, Adv Mater for example, and 2005,17 (4): the 412-415 disclosed method prepares.
Wherein add the aqueous solution of sulfide in the step (2), make that the mol ratio of triangle silver nano-grain and sulfide is 50: 1-5: 1.
Wherein used sulfide is vulcanized sodium or potassium sulfide in the step (2).
In addition, the present invention also provides the application process of this New triangular silver-silver sulfide Nano composite granules in the preparation oligonucleotide probe, may further comprise the steps:
(1) triangle Ag-Ag
2The S Nano composite granules washs with phosphate buffer;
(2) under the room temperature, with triangle Ag-Ag
2The S Nano composite granules mixes the centrifugal supernatant of removing with the oligonucleotides (HS-DNA) and the borate buffer of sulfydryl modification;
(3) deposition obtains triangle Ag-Ag with the phosphate buffer washing
2S Nano composite granules oligonucleotide probe;
(4) the triangle Ag-Ag that obtains
2S Nano composite granules oligonucleotide probe heavily is distributed in the phosphate buffer, in 4 ℃ of storages.
Wherein, the pH value of the used phosphate buffer of step (1) is 7.5-8.5, contains the NaCl that concentration is 0.1-0.2mol/L; Preferred pH value is 8.0, contains the NaCl that concentration is 0.15mol/L.
Wherein, the pH value of used phosphate buffer is 6.5-7.5 in step (3) and (4), contains the NaCl that concentration is 0.1-0.2mol/L; Preferred pH value is 7.0, contains the NaCl that concentration is 0.15mol/L.
Wherein, the pH value of used borate buffer is 8.5-9.5 in the step (2), and concentration is 45-55mmol/L; Preferred pH value is 9.2, and concentration is 50mmol/L.
The invention has the advantages that:
(1) a kind of simple triangle Ag-Ag of synthesizing new fast is provided
2The preparation method of S Nano composite granules;
(2) through 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
2The S Nano composite granules prepares the process of oligonucleotide probe and has 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 synthesizes
2The S Nano composite granules can be used for the detection of dna molecular, and it is with a wide range of applications in the bio-sensing field as the novel nano bioprobe.
Description of drawings
Fig. 1 is triangle Ag-Ag of the present invention
2S Nano composite granules oligonucleotide probe prepares sketch map.
Fig. 2 is the triangle Ag-Ag of one embodiment of the invention preparation
2The picture of the transmission electron microscope of S Nano composite granules (TEM).
Fig. 3 is the triangle Ag-Ag of one embodiment of the invention preparation
2The melting temperature sketch map of S nano particle oligonucleotide probe and target DNA.
Fig. 4 is the triangle Ag-Ag of further embodiment of this invention preparation
2The picture of the transmission electron microscope of S Nano composite granules (TEM).
Fig. 5 is the triangle Ag-Ag of further embodiment of this invention preparation
2The melting temperature sketch map of S nano particle oligonucleotide probe and target DNA.
Fig. 6 is the triangle Ag-Ag of another embodiment of the present invention preparation
2The picture of the transmission electron microscope of S Nano composite granules (TEM).
Fig. 7 is the triangle Ag-Ag2S nano particle oligonucleotide probe of another embodiment of the present invention preparation and the melting temperature sketch map of target DNA.
The specific embodiment
Embodiment 1:
Triangle Ag-Ag
2The preparation of S Nano composite granules:
(1) (Adv Mater, 2005,17 (4): method 412-415.) prepares the triangle silver nano-grain of PVP protection to reference literature: use PVP and citrate as stabilizing agent and masterplate agent, use NaBH under the lucifuge stirring condition
4Reduction AgNO
3Prepare the triangle silver nano-grain of PVP protection.
(2) in the triangle silver nano-grain solution of step (1) preparation, add 1 μ L1mmol/L sodium sulfide solution under the lucifuge condition fast, the molar ratio of triangle silver nano-grain and vulcanized sodium is 10: 1 approximately.Constant temperature stirred 30 minutes for 25 ℃, obtained triangle Ag-Ag
2The S Nano composite granules.Its pattern is seen accompanying drawing 2.
Utilize triangle Ag-Ag
2The S Nano composite granules prepares oligonucleotide probe and cross performance test case thereof:
The model oligonucleotide molecules is following:
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 for preparing by Fig. 1
2The S Nano composite granules washs with phosphate buffer (PBS) (pH 8.0, NaCl 0.15mol/L).
(2) under the room temperature condition, with 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 deposition is cleaned with phosphate buffer (pH 7.0, NaCl 0.15mol/L).The triangle Ag-Ag that the sulfydryl that obtains is oligonucleotides-modified
2(nano-probe a) heavily is distributed in the phosphate buffer and in 4 ℃ of storages the S nano particle.
(3) nano-probe b is through same method and probe b prepared in reaction, heavily is scattered in the phosphate buffer and in 4 ℃ of storages.
(4) nano-probe a and b are mixed with target DNA, be heated to more than 70 ℃ 10 minutes, cooling naturally uses spectrophotometer that its melting temperature is detected.See accompanying drawing 3.
Embodiment 2:
Triangle Ag-Ag
2The preparation of S Nano composite granules:
Preparation method and instance 1 are basic identical; Difference is: in the triangle silver nano-grain solution of step 1) preparation, add the sodium sulfide solution of 3 μ L 2mmol/L in the step (2) under the lucifuge condition fast, the molar ratio of triangle silver nano-grain and vulcanized sodium is 20: 1 approximately.Constant temperature stirred 15 minutes for 30 ℃, obtained triangle Ag-Ag after the reaction
2The S Nano composite granules.Its pattern is seen accompanying drawing 4.
Utilize triangle Ag-Ag
2The S Nano composite granules prepares oligonucleotide probe and cross performance test case thereof:
The model oligonucleotide molecules is following:
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 for preparing by Fig. 4
2The S Nano composite granules washs with phosphate buffer (PBS) (pH 8.0, NaCl 0.15mol/L).
(2) under the room temperature condition, with 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 deposition is cleaned with phosphate buffer (pH 7.0, NaCl 0.15mol/L).The triangle Ag-Ag that the sulfydryl that obtains is oligonucleotides-modified
2(nano-probe a) heavily is distributed in the phosphate buffer and in 4 ℃ of storages the S nano particle.
(3) nano-probe b is through same method and probe b prepared in reaction, heavily is scattered in the phosphate buffer and in 4 ℃ of storages.
(4) nano-probe a and b are mixed with target DNA, be heated to more than 60 ℃ 10 minutes, cooling naturally uses spectrophotometer that its melting temperature is detected.See accompanying drawing 5.
Embodiment 3:
Triangle Ag-Ag
2The preparation of S Nano composite granules:
Preparation method and instance 1 are basic identical; Difference is: in the triangle silver nano-grain solution of step 1) preparation, add the potassium sulfide aqueous solution of 2 μ L 3mmol/L in the step (2) under the lucifuge condition fast, the molar ratio of triangle silver nano-grain and potassium sulfide is 30: 1 approximately.Constant temperature stirred 25 minutes for 20 ℃, obtained triangle Ag-Ag after the reaction
2The S Nano composite granules.Its pattern is seen accompanying drawing 6.
Utilize triangle Ag-Ag
2The S Nano composite granules prepares oligonucleotide probe and cross performance test case thereof:
The model oligonucleotide molecules is following:
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 for preparing by Fig. 6
2The S Nano composite granules washs with phosphate buffer (PBS) (pH 8.0, NaCl 0.15mol/L).
(2) under the room temperature condition, with 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 deposition is cleaned with phosphate buffer (pH 7.0, NaCl 0.15mol/L).The triangle Ag-Ag that the sulfydryl that obtains is oligonucleotides-modified
2(nano-probe a) heavily is distributed in the phosphate buffer and in 4 ℃ of storages the S nano particle.
(3) nano-probe b is through with quadrat method and probe b prepared in reaction, heavily is scattered in the phosphate buffer and in 4 ℃ of storages.
(4) nano-probe a and b are mixed with target DNA, be heated to more than 80 ℃ 5 minutes, cooling naturally uses spectrophotometer that its melting temperature is detected.See accompanying drawing 7.
The triangle Ag-Ag that embodiment of the invention 1-3 prepares
2S Nano composite granules oligonucleotide probe is that the oligonucleotide probe that precursor prepares has preparation easy than what reported with three horn silver AgXs; The characteristics that repeat easily; And from the melting temperature curve of accompanying drawing 3,5,7, can obviously find out: the sudden change through the monitoring absorbance just can obtain melting temperature numerical value very accurately, and this this probe of explanation will be a kind of very desirable material in following oligonucleotides context of detection.
Claims (10)
1. triangle silver-silver sulfide Nano composite granules; Wherein comprise the triangle silver nano-grain; It is characterized in that: at the protective layer that the surface coverage of triangle silver nano-grain has silver sulfide to form, said silver sulfide is by triangle silver nano-grain and the direct prepared in reaction of sulfide.
2. triangle silver as claimed in claim 1-silver sulfide Nano composite granules, it is characterized in that described silver sulfide can be directly with the oligonucleotides of sulfydryl modification mutually coupling prepare corresponding oligonucleotide probe.
3. method for preparing triangle silver-silver sulfide Nano composite granules according to claim 1 or claim 2,, it is characterized in that mainly may further comprise the steps:
(1) the triangle silver nano-grain of preparation polyvinylpyrrolidone (PVP) protection;
(2) in the solution of step (1), add fast the aqueous solution of sulfide under the lucifuge condition, constant temperature keeps 20-35 ℃, stirs 5-30 minute.
4. preparation method as claimed in claim 3 is characterized in that: the described triangle silver nano-grain of step (1) is by existing method (Adv Mater, 2005,17 (4): 412-415.) prepared.
5. preparation method as claimed in claim 3 is characterized in that: the aqueous solution of the sulfide that adds in the said step (2) makes that the mol ratio of triangle silver nano-grain and sulfide is 50: 1-5: 1.
6. preparation method as claimed in claim 3 is characterized in that: used sulfide is vulcanized sodium or potassium sulfide in the said step (2).
7. the application of each described triangle silver-silver sulfide Nano composite granules of claim 1-6 is that it is prepared corresponding oligonucleotide probe, may further comprise the steps:
(1) triangle Ag-Ag
2The S Nano composite granules washs with phosphate buffer;
(2) under the room temperature, with triangle Ag-Ag
2The S Nano composite granules mixes with the oligonucleotides and the borate buffer of sulfydryl modification, the centrifugal supernatant of removing;
(3) deposition obtains triangle Ag-Ag with the phosphate buffer washing
2S Nano composite granules oligonucleotide probe;
(4) the triangle Ag-Ag that obtains
2S Nano composite granules oligonucleotide probe heavily is distributed in the phosphate buffer, in 4 ℃ of storages.
8. application as claimed in claim 7 is characterized in that, the pH value of the used phosphate buffer of step (1) is 7.5-8.5, contains the NaCl that concentration is 0.1-0.2mol/L; Preferred pH value is 8.0, contains the NaCl that concentration is 0.15mol/L.
9. application as claimed in claim 7 is characterized in that, the pH value of used phosphate buffer is 6.5-7.5 in step (3) and (4), contains the NaCl that concentration is 0.1-0.2mol/L; Preferred pH value is 7.0, contains the NaCl that concentration is 0.15mol/L.
10. application as claimed in claim 7 is characterized in that, the pH value of used borate buffer is 8.5-9.5 in the step (2), and concentration is 45-55mmol/L; Preferred pH value is 9.2, and concentration is 50mmol/L.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103785848A (en) * | 2013-12-26 | 2014-05-14 | 江苏大学 | Method for using single tank to synthesize Ag-Ag2S/CdS heterogeneous structure |
| CN105412940A (en) * | 2015-12-02 | 2016-03-23 | 鲁东大学 | Composite nanometer antibacterial material used for treating vancomycin drug resistant pathogenic bacteria |
| CN107159881A (en) * | 2017-06-12 | 2017-09-15 | 青岛科技大学 | A kind of silver-colored@silver sulfides core shell structure photothermal deformation nano material |
| 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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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103785848A (en) * | 2013-12-26 | 2014-05-14 | 江苏大学 | Method for using single tank to synthesize Ag-Ag2S/CdS heterogeneous structure |
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| 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 |
| CN107159881A (en) * | 2017-06-12 | 2017-09-15 | 青岛科技大学 | A kind of silver-colored@silver sulfides core shell structure photothermal deformation nano material |
| 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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