CN104923777A - High-salt-tolerance metal nanoparticle assembly and preparing method thereof - Google Patents
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Abstract
The invention discloses a high-salt-tolerance metal nanoparticle assembly and a preparing method thereof. The method comprises the steps that metal nanoparticles a and ssDNA are taken to be added into mixed liquor a of a wrapping agent, a buffered solution and salt, and after mixed cultivation is carried out, separation and purification are carried out, and a product a is obtained; metal nanoparticles b and sscDNA are taken to be added into mixed liquor b of a wrapping agent, a buffered solution and salt, and after mixed cultivation is carried out, separation and purification are carried out, and a product b is obtained; and the product a and the product b are mixed and then dissolved into mixed liquor of a wrapping agent, a buffered solution and salt, and after stir cultivation is carried out, centrifugal washing is carried out, and the high-salt-tolerance metal nanoparticle assembly is obtained. The preparing method is simple, efficient and high in controllability. The obtained assembly is uniform in morphology and good in dispersibility. More significantly, the assembly prepared through the method has the high salt tolerance and can be expected to serve as a nano carrier to be applied to the fields like the biomedicine.
Description
Technical field
The invention belongs to technical field of nano material, be specifically related to a kind of high-salt tolerance metal nanoparticle assembly and preparation method thereof.
Background technology
In recent years, metal nanoparticle assembly achieves and applies widely in nano-device, nano-sensor and nanosecond medical science etc.Metal nano assembly not only has the performance such as optics, electricity, catalysis of isolated nano particle excellence, and make assembly have some new characteristics owing to assembling, as surface-enhanced Raman characteristic, this has greatly widened the range of application of metal nanoparticle in field of nanometer technology.And along with the application of metal nanoparticle assembly more and more extensive, particularly at biomedical aspect, very high to the requirement of its salt tolerance.Metal nanoparticle, comprises metal nanoparticle assembly, is all colloid, to electrolyte solution as salting liquid is responsive especially.Adding a small amount of salt just easily makes nano particle reunite, and organism inner salt is dense, therefore, metal nanoparticle assembly is applied in biomedical sector, must prepare a kind of assembly of high-salt tolerance.
The domestic patent of invention about high-salt tolerance metal nanoparticle assembly or blank at present.In fact, the technological invention about isolated salt tolerance metal nanoparticle is also very few for it.Application number be the Chinese invention patent " preparation method of the golden nanometer particle of a kind of high stability and functionalization " of 201010286955.X disclose a kind of DNA of utilization assist synthesis golden nanometer particle method.Golden nanometer particle regular shape, uniform particle diameter, salt tolerance prepared by the method can be enhanced to 20 times, but the method is only for isolated golden nanometer particle.
Summary of the invention
In order to overcome the shortcoming of prior art with not enough, primary and foremost purpose of the present invention is to provide a kind of preparation method with the metal nanoparticle assembly of high-salt tolerance, and this preparation method is simple efficiently, product morphology is even, good dispersion.
Another object of the present invention is to provide the metal nanoparticle assembly adopting above-mentioned preparation method to obtain, and carry out the salt tolerance experiment of assembly.
Object of the present invention is achieved through the following technical solutions:
A preparation method for high-salt tolerance metal nanoparticle assembly, comprises the steps:
(1) get metal nanoparticle a and ssDNA, join in the mixed liquor a of coating agent, cushioning liquid and salt, separation and purification after Mixed culture, obtain product a;
(2) get metal nanoparticle b and sscDNA, join in the mixed liquor b of coating agent, cushioning liquid and salt, separation and purification after Mixed culture, obtain product b;
(3), after the product a of step (1) and (2) and product b being mixed, be dissolved in the mixed solution of coating agent, cushioning liquid and salt, centrifuge washing after stir culture, obtain high-salt tolerance metal nanoparticle assembly.
In said method, the metal nanoparticle a described in step (1) is one or both in Au, Ag or Cu; The size of described metal nanoparticle is 1 ~ 100 nm; The mol ratio of described metal nanoparticle a and ssDNA is 1:40 ~ 400.
In said method, the coating agent described in step (1) is lauryl sodium sulfate (SDS); Described coating agent is 0.01 % ~ 1 % with the mass ratio of the mixed liquor a adding metal nanoparticle a and ssDNA; Described cushioning liquid is phosphate buffer (PBS), pH=7.3, and the concentration of described cushioning liquid in the mixed liquor a adding metal nanoparticle a and ssDNA is 10 ~ 100 mmol/L; The concentration of described salt in the mixed liquor a adding metal nanoparticle a and ssDNA progressively increases to final 50 ~ 500 mmol/L in 6 ~ 24 h.
In said method, the time of the Mixed culture described in step (1) is 6 ~ 24 h; The mode of described separation and purification is centrifugation or dialysis.
In said method, the metal nanoparticle a in the metal nanoparticle b described in step (2) and step (1) is identical or different, and described metal nanoparticle b is one or both in Au, Ag or Cu; The size of described metal nanoparticle b is 1 ~ 100 nm; The mol ratio of described metal nanoparticle b and sscDNA is 1:0.2 ~ 20.
In said method, the coating agent described in step (2) is SDS, and described coating agent is 0.01 % ~ 1 % with the mass ratio of the mixed liquor b adding metal nanoparticle b and sscDNA; Described cushioning liquid is PBS, pH=7.3, and the concentration of described cushioning liquid in the mixed liquor b adding metal nanoparticle b and sscDNA is 10 ~ 100 mmol/L; The concentration of described salt in the mixed liquor b adding metal nanoparticle b and sscDNA is 50 ~ 500 mmol/L.
In said method, the time of the Mixed culture described in step (2) is 6 ~ 24 h; Described separation and purification mode is centrifugation or dialysis.
In said method, the product a described in step (3) and the mol ratio of product b are 1:10 ~ 400; Described coating agent is SDS; Described coating agent is 0.01% ~ 1 % with the mass ratio of the mixed solution being mixed with product a and product b; Described cushioning liquid is PBS, pH=7.3; The concentration of described cushioning liquid in the mixed solution being mixed with product a and product b is 10 ~ 100 mmol/L; The concentration of described salt in the mixed solution being mixed with product a and product b is 50 ~ 500 mmol/L; The described Mixed culture time is 6 ~ 24 h; Described centrifugal rotational speed is 5000 ~ 20,000 rpm; Centrifugation time is 5 ~ 30 min, centrifugal 1 ~ 3 time.
The salt tolerance experiment of above-mentioned metal nanoparticle assembly, is included in the stability in physiological saline and the stability in variable concentrations sodium chloride (NaCl) solution.
The present invention has following advantage and effect relative to prior art:
The method is easy to operation, controllability is strong, packaging efficiency is high, and the structure of products therefrom is homogeneous, good dispersion, and product has very strong salt resistant character, can bear the salinity up to physiological saline concentration three times.Therefore, the metal nano assembly that this preparation method obtains has potential advantage in biologic applications.
Accompanying drawing explanation
Fig. 1 is the transmission electron microscope figure (TEM) of the golden nanometer particle assembly of embodiment 2 gained;
Fig. 2 is the transmission electron microscope figure (TEM) after the golden nanometer particle assembly of embodiment 2 gained cultivates 12h in physiological saline;
Fig. 3 is the salt tolerance experimental result of golden nanometer particle assembly in variable concentrations NaCl solution of embodiment 2 gained, i.e. color variation diagram;
Fig. 4 be the golden nanometer particle assembly of embodiment 2 gained in variable concentrations NaCl solution salt tolerance experimental result, i.e. uv absorption spectra.
Detailed description of the invention
Below by embodiment and accompanying drawing, this technological invention is further described, but embodiments of the present invention are not limited only to this.
Metal nanoparticle used in the inventive method can, from buying on the market, also can adopt prior art to prepare voluntarily.
SsDNA and sscDNA that following examples use is purchased from Shanghai Sheng Gong bioengineering limited company.
Its sequence number is as follows:
ssDNA:5’-HS-(CH
2)
6 ATC CTG ACA TCG GCA CGA GTA TTT CTA CCA TGT ATC-3’
sscDNA:5’-HS-(CH
2)
6 GAT ACA TGG TAG AAA TAC TCGTGC CGA TGT CAG GAT-3’。
embodiment 1
(1) by 18 nm golden nanometer particles and the ssDNA mol ratio according to 1:100, join ultimate density (namely relative to the concentration adding the mixed liquor having golden nanometer particle and ssDNA, lower same) be in the mixed liquor a of 0.2 % (mass fraction) SDS, 20 mM PBS (pH=7.3) and 300 mM NaCl, after Mixed culture 20 h, centrifugal 3 times of 15000 rpm, each 15 min.Get lower sediment;
(2) by 4 nm golden nanometer particles and the sscDNA molar ratio according to 1:10, joining ultimate density is in the mixed liquor b of 0.2 % (mass fraction) SDS, 20 mM PBS (pH=7.3) and 100 mM NaCl, dialyses 20 h with purifying after Mixed culture 6 h with bag filter;
(3) by the product of step (1) and (2) according to after the mixed in molar ratio of 1:20, joining ultimate density is in the mixed solution of 0.2 % (mass fraction) SDS, 20 mM PBS (pH=7.3) and 150 mM NaCl, slow stir culture 20 h, then 15, under 000 rpm centrifugal 3 times, each 15 min, obtain golden nanometer particle assembly.
embodiment 2
(1) by 18 nm golden nanometer particles and the ssDNA mol ratio according to 1:100, joining ultimate density is in the mixed liquor a of 0.2 % (mass fraction) SDS, 20 mM PBS (pH=7.3) and 300 mM NaCl, after Mixed culture 20 h, 1, centrifugal 3 times of 5000 rpm, each 15 min.Get lower sediment;
(2) by 4 nm golden nanometer particles and the sscDNA molar ratio according to 1:10, joining ultimate density is in the mixed liquor b of 0.2 % SDS, 20 mM PBS (pH=7.3) and 100 mM NaCl, dialyses 20 h with purifying after Mixed culture 6 h with bag filter;
(3) by the product of step (1) and (2) according to after the mixed in molar ratio of 1:40, joining ultimate density is in the mixed solution of 0.2 % SDS, 20 mM PBS (pH=7.3) and 150 mM NaCl, slow stir culture 20 h, then 15, under 000 rpm centrifugal 3 times, each 15 min, obtain golden nanometer particle assembly.
Carry out TEM sign to the golden nanometer particle assembly of gained, result as shown in Figure 1.The golden nanometer particle of 4 nm is successfully connected to 18 nm golden nanometer particles surfaces as can be seen from Figure 1, forms " core-satellite " shape assembly, satellite " mean number be 9, and the good dispersion of assembly, size is more even.
embodiment 3
(1) by 18 nm golden nanometer particles and the ssDNA mol ratio according to 1:100, joining ultimate density is in the mixed liquor a of 0.2 % (mass fraction) SDS, 20 mM PBS (pH=7.3) and 300 mM NaCl, after Mixed culture 20 h, 1, centrifugal 3 times of 5000 rpm, each 15 min.Get lower sediment;
(2) by 4 nm golden nanometer particles and sscDNA according to the molar ratio of 1:10, join in the mixed liquor b of 0.2 % SDS, 20 mM PBS (pH=7.3) and 100 mM NaCl, after Mixed culture 6 h, dialyse 20 h with purifying with bag filter;
(3) by the product of step (1) and (2) according to after the mixed in molar ratio of 1:80, joining ultimate density is in the mixed liquor of 0.2 % SDS, 20 mM PBS (pH=7.3) and 150 mM NaCl, slow stir culture 20 h, then 15, under 000 rpm centrifugal 3 times, each 15 min, obtain golden nanometer particle assembly.
embodiment 4
(1) by 18 nm golden nanometer particles and the ssDNA mol ratio according to 1:100, joining ultimate density is in the mixed liquor a of 0.2 % (mass fraction) SDS, 20 mM PBS (pH=7.3) and 300 mM NaCl, after Mixed culture 20 h, 1, centrifugal 3 times of 5000 rpm, each 15 min.Get lower sediment;
(2) by 4 nm golden nanometer particles and sscDNA according to the molar ratio of 1:10, join in the mixed liquor b of 0.2 % SDS, 20 mM PBS (pH=7.3) and 100 mM NaCl, after Mixed culture 6 h, dialyse 20 h with purifying with bag filter;
(3) by the product of step (1) and (2) according to after the mixed in molar ratio of 1:160, joining ultimate density is in the mixed solution of 0.2 % SDS, 20 mM PBS (pH=7.3) and 150 mM NaCl, slow stir culture 20 h, then 15, under 000 rpm centrifugal 3 times, each 15 min, obtain golden nanometer particle assembly.
salt resistant character is tested
(1), after the golden nanometer particle assembly of Example 2 cultivates 12 h in physiological saline, carry out transmission electron microscope (TEM) and characterize.Its TEM schemes as shown in Figure 2.
As can be seen from Figure 2, golden nanometer particle assembly prepared by the inventive method still keeps complete pattern after cultivating in physiological saline and reaching 12 h, illustrates that it has good stability in the salinity of 0.9 %.
(2) the golden nanometer particle assembly of Example 2 is respectively at 0.9 %, 1.8 %, 3.0 %, 3.3 % and 4.5 %(mass fractions) NaCl solution in carry out salt tolerance test, observe the initial color of solution and change and carry out ultraviolet sign.Result as shown in Figure 3 and Figure 4.
The stability of golden nanometer particle assembly is easy to the impact being subject to salinity.The normal homodisperse assembly aqueous solution generally in ruby red, and may produce agglomeration when it is in high concentration salt solutions.This is reflected in color change (usually becoming purple) being solution in appearance, is reflected in the red shift being maximum absorption band in the middle of ultraviolet spectra.Therefore its stability can be verified by solution colour and ultra-violet absorption spectrum.The color change display of Fig. 3: in the NaCl solution (being equivalent to the concentration of physiological saline) of 0.9 %, not there is agglomeration in the golden nanometer particle assembly of embodiment 2 gained, color is initial ruby red, and the result that this and salt tolerance test (1) is coincide mutually; When salinity increase about twice time (3.0 %), still do not have agglomeration to occur, solution is still ruby red, shows that assembly now still has good stability.As can be seen from Figure 4, when salinity is up to 3.0 %, the maximum absorption band of golden nanometer particle assembly does not still have obvious red shift, and this is also consistent with the color result of variations of Fig. 3.Illustrate that product has very high salt resistant character, is expected to apply under physiological environment thus.
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from Spirit Essence of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (9)
1. a preparation method for high-salt tolerance metal nanoparticle assembly, is characterized in that, comprises the steps:
(1) get metal nanoparticle a and ssDNA, join in the mixed liquor a of coating agent, cushioning liquid and salt, separation and purification after Mixed culture, obtain product a;
(2) get metal nanoparticle b and sscDNA, join in the mixed liquor b of coating agent, cushioning liquid and salt, separation and purification after Mixed culture, obtain product b;
(3), after the product a of step (1) and (2) and product b being mixed, be dissolved in the mixed solution of coating agent, cushioning liquid and salt, centrifuge washing after stir culture, obtain high-salt tolerance metal nanoparticle assembly.
2. the preparation method of a kind of high-salt tolerance metal nanoparticle assembly according to claim 1, is characterized in that: the metal nanoparticle a described in step (1) is one or both in Au, Ag or Cu; The size of described metal nanoparticle is 1 ~ 100 nm; The mol ratio of described metal nanoparticle a and ssDNA is 1:40 ~ 400.
3. the preparation method of a kind of high-salt tolerance metal nanoparticle assembly according to claim 1, is characterized in that: the coating agent described in step (1) is lauryl sodium sulfate (SDS); Described coating agent is 0.01 % ~ 1 % with the mass ratio of the mixed liquor a adding metal nanoparticle a and ssDNA; Described cushioning liquid is phosphate buffer (PBS), pH=7.3, and the concentration of described cushioning liquid in the mixed liquor a adding metal nanoparticle a and ssDNA is 10 ~ 100 mmol/L; The concentration of described salt in the mixed liquor a adding metal nanoparticle a and ssDNA increases to final 50 ~ 500 mmol/L in 6 ~ 24 h.
4. the preparation method of a kind of high-salt tolerance metal nanoparticle assembly according to claim 1, is characterized in that: the time of the Mixed culture described in step (1) is 6 ~ 24 h; The mode of described separation and purification is centrifugation or dialysis.
5. the preparation method of a kind of high-salt tolerance metal nanoparticle assembly according to claim 1, it is characterized in that: the metal nanoparticle a in the metal nanoparticle b described in step (2) and step (1) is identical or different, described metal nanoparticle b is one or both in Au, Ag or Cu; The size of described metal nanoparticle b is 1 ~ 100 nm; The mol ratio of described metal nanoparticle b and sscDNA is 1:0.2 ~ 20.
6. the preparation method of a kind of high-salt tolerance metal nanoparticle assembly according to claim 1, it is characterized in that: the coating agent described in step (2) is SDS, described coating agent is 0.01 % ~ 1 % with the mass ratio of the mixed liquor b adding metal nanoparticle b and sscDNA; Described cushioning liquid is PBS, pH=7.3, and the concentration of described cushioning liquid in the mixed liquor b adding metal nanoparticle b and sscDNA is 10 ~ 100 mmol/L; The concentration of described salt in the mixed liquor b adding metal nanoparticle b and sscDNA is 50 ~ 500 mmol/L.
7. the preparation method of a kind of high-salt tolerance metal nanoparticle assembly according to claim 1, is characterized in that: the time of the Mixed culture described in step (2) is 6 ~ 24 h; Described separation and purification mode is centrifugation or dialysis.
8. the preparation method of a kind of high-salt tolerance metal nanoparticle assembly according to claim 1, is characterized in that: the product a described in step (3) and the mol ratio of product b are 1:10 ~ 400; Described coating agent is SDS; Described coating agent is 0.01% ~ 1 % with the mass ratio of the mixed solution being mixed with product a and product b; Described cushioning liquid is PBS, pH=7.3; The concentration of described cushioning liquid in the mixed solution being mixed with product a and product b is 10 ~ 100 mmol/L; The concentration of described salt in the mixed solution being mixed with product a and product b is 50 ~ 500 mmol/L; The described Mixed culture time is 6 ~ 24 h; Described centrifugal rotational speed is 5000 ~ 20,000 rpm; Centrifugation time is 5 ~ 30 min, centrifugal 1 ~ 3 time.
9. according to any one of claim 1 to 8, preparation method prepares high-salt tolerance metal nanoparticle assembly.
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