WO2016145925A1 - High salt-resistant metal nanoparticle assembly and manufacturing method thereof - Google Patents
High salt-resistant metal nanoparticle assembly and manufacturing method thereof Download PDFInfo
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- WO2016145925A1 WO2016145925A1 PCT/CN2015/100036 CN2015100036W WO2016145925A1 WO 2016145925 A1 WO2016145925 A1 WO 2016145925A1 CN 2015100036 W CN2015100036 W CN 2015100036W WO 2016145925 A1 WO2016145925 A1 WO 2016145925A1
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B22—CASTING; POWDER METALLURGY
- B22F—WORKING METALLIC POWDER; MANUFACTURE OF ARTICLES FROM METALLIC POWDER; MAKING METALLIC POWDER; APPARATUS OR DEVICES SPECIALLY ADAPTED FOR METALLIC POWDER
- B22F1/00—Metallic powder; Treatment of metallic powder, e.g. to facilitate working or to improve properties
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
Definitions
- the invention belongs to the technical field of nano materials, and particularly relates to a high salt tolerance metal nanoparticle assembly and a preparation method thereof.
- Metal nanoparticle assemblies have found wide applications in nanodevices, nanosensors, and nanomedicine.
- Metal nano-assemblies not only have excellent optical, electrical, catalytic and other properties of isolated nanoparticles, but also have some new properties, such as surface-enhanced Raman properties, due to assembly, which greatly broadens the field of nano-particles in nanotechnology.
- the scope of application within. With the increasing application of metal nanoparticle assemblies, especially in biomedicine, the salt tolerance is very high.
- Metal nanoparticles, including metal nanoparticle assemblies are colloids that are particularly sensitive to electrolyte solutions such as salt solutions.
- the primary object of the present invention is to provide a method for preparing a metal nanoparticle assembly having high salt tolerance, which is simple and efficient, has uniform product morphology, and good dispersibility.
- Another object of the present invention is to provide a metal nanoparticle assembly obtained by the above production method, and to carry out an experiment for salt tolerance of the assembly.
- a method for preparing a high salt tolerance metal nanoparticle assembly comprising the following steps:
- the metal nanoparticle a according to the step (1) is Au, Ag or Cu.
- the coating agent in the step (1) is sodium dodecyl sulfate (SDS); the coating agent and the metal nanoparticles are added
- SDS sodium dodecyl sulfate
- the mass ratio of a to ssDNA mixture a is 0.01% ⁇ 1%
- PBS phosphate buffered saline
- the buffer solution has a concentration of 10 to 100 mmol/L in a mixture a of metal nanoparticles a and ssDNA; the salt is added with metal nanoparticles a and
- the concentration in the mixture of ssDNA a gradually increased from 6 to 24 h to the final 50 to 500 mmol/L.
- the mixing culture time described in the step (1) is 6 to 24 h.
- the separation and purification method is centrifugal separation or dialysis.
- the metal nanoparticles b described in the step (2) and the metal nanoparticles in the step (1) are one or two of Au, Ag or Cu; the metal nanoparticles b have a size of 1 to 100 nm; the metal nanoparticles b
- the molar ratio to sscDNA is 1:0.2 ⁇ 20.
- the encapsulating agent in the step (2) is SDS, and the wrapping agent is added with metal nanoparticles b and
- the mass ratio of the mixture b of sscDNA is 0.01% ⁇ 1%
- the concentration of the mixture of sscDNA b is 10 ⁇ 100 mmol/L
- the concentration of the salt in the mixture b of metal nanoparticles b and sscDNA is 50-500. Mmmol/L ;
- the mixing culture time described in the step (2) is 6 to 24 h.
- the separation and purification method is centrifugal separation or dialysis.
- the concentration of the buffer solution in the mixed solution in which the product a and the product b are mixed is 10 to 100 mmol/L; the concentration of the salt in the mixed solution in which the product a and the product b are mixed is 50 ⁇ 500 mmol/L;
- the mixed culture time is 6 ⁇ 24 h; the centrifugal speed is 5000 ⁇ 20,000 rpm; the centrifugation time is 5 ⁇ 30 min, centrifugation 1 ⁇ 3 times.
- Salt tolerance test of the above metal nanoparticle assembly including stability in physiological saline and sodium chloride at different concentrations ( NaCl Stability in solution.
- the method is easy to operate, has strong controllability and high assembly efficiency, and the obtained product has uniform structure and good dispersibility, and the product has strong salt tolerance and can withstand a salt concentration up to three times the concentration of physiological saline. Therefore, the metal nanoassemblies obtained by the preparation method have potential advantages in biological applications.
- Figure 1 is a transmission electron micrograph (TEM) of the gold nanoparticle assembly obtained in Example 2;
- Figure 2 is a transmission electron micrograph of the gold nanoparticle assembly obtained in Example 2 after incubation for 12 h in physiological saline ( TEM );
- Figure 3 is a gold nanoparticle assembly obtained in Example 2 at different concentrations of NaCl
- the salt tolerance test result in the solution, that is, the ultraviolet absorption spectrum.
- the metal nanoparticles used in the method of the present invention are commercially available or can be prepared by themselves using the prior art.
- the ssDNA and sscDNA used in the following examples were purchased from Shanghai Shenggong Bioengineering Co., Ltd.
- 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'.
- steps (1) and (2) are mixed at a molar ratio of 1:40 and added to a final concentration of 0.2.
- the obtained gold nanoparticle assembly was subjected to TEM characterization, and the results are shown in Fig. 1.
- the gold nanoparticles were successfully connected to the surface of 18 nm gold nanoparticles to form a 'nuclear-satellite' assembly.
- the average number of satellites was nine, and the assembly was well dispersed and uniform in size.
- steps (1) and (2) are mixed at a molar ratio of 1:80 and added to a final concentration of 0.2.
- Example 2 The gold nanoparticle assembly of Example 2 was cultured in physiological saline for 12 hours, and then subjected to transmission electron microscopy (TEM). ) characterization. Its TEM image is shown in Figure 2.
- TEM transmission electron microscopy
- the gold nanoparticle assembly prepared by the method of the present invention is cultured in physiological saline for up to 12 h. It still retains its intact morphology, indicating its good stability at 0.9% salt concentration.
- Example 2 The gold nanoparticle assembly of Example 2 was taken at 0.9%, 1.8%, 3.0%, 3.3, respectively. Salt tolerance tests were performed in % and 4.5% (mass fraction) NaCl solutions, and the initial color change of the solution was observed and UV characterized (results shown in Figure 3).
- the stability of the gold nanoparticle assembly is easily affected by the salt concentration.
- a normally uniformly dispersed aqueous solution of the assembly is generally ruby red, and agglomeration may occur when it is in a high concentration of salt solution. This is reflected in the appearance of the color change of the solution (usually purple), which is reflected in the ultraviolet spectrum as the red shift of the maximum absorption peak. Therefore, the stability of the solution can be verified by the color of the solution and the ultraviolet absorption spectrum.
Abstract
Description
Claims (9)
- 一种高耐盐性金属纳米粒子组装体的制备方法,其特征在于,包括如下步骤:A method for preparing a high salt tolerance metal nanoparticle assembly, comprising the steps of:(1)取金属纳米粒子a与ssDNA,加入到包裹剂、缓冲溶液和盐的混合液a中,混合培养后分离纯化,得到产物a;(1) taking metal nanoparticles a and ssDNA, added to a mixture of a coating agent, a buffer solution and a salt, mixed and cultured, and then isolated and purified to obtain a product a;(2)取金属纳米粒子b与sscDNA,加入到包裹剂、缓冲溶液和盐的混合液b中,混合培养后分离纯化,得到产物b;(2) taking metal nanoparticles b and sscDNA, added to the mixture b of the package, buffer solution and salt, mixed culture and separation and purification, to obtain product b;(3)将步骤(1)和(2)的产物a和产物b混合后,溶解于包裹剂、缓冲溶液和盐的混合溶液中,搅拌培养后离心洗涤,得到高耐盐性金属纳米粒子组装体。(3) mixing the product a and the product b of the steps (1) and (2), dissolving in a mixed solution of a coating agent, a buffer solution and a salt, stirring and culturing, and then centrifuging to obtain a high salt-tolerant metal nanoparticle assembly. body.
- 根据权利要求1所述的一种高耐盐性金属纳米粒子组装体的制备方法,其特征在于:步骤(1)所述的金属纳米粒子a为Au、Ag或Cu中的一种或两种;所述的金属纳米粒子的尺寸是1~100 nm;所述的金属纳米粒子a与ssDNA的摩尔比为1:40~400。 The method for preparing a high salt-tolerant metal nanoparticle assembly according to claim 1, wherein the metal nanoparticle a according to the step (1) is one or two of Au, Ag or Cu. The size of the metal nanoparticles is 1~100 Nm; the molar ratio of the metal nanoparticles a to ssDNA is 1:40-400.
- 根据权利要求1所述的一种高耐盐性金属纳米粒子组装体的制备方法,其特征在于:步骤(1)所述的包裹剂为十二烷基硫酸钠(SDS);所述包裹剂与加入了金属纳米粒子a与ssDNA的混合液a的质量比为0.01 %~1 %;所述的缓冲溶液为磷酸缓冲液(PBS),pH= 7.3,所述缓冲溶液在加入了金属纳米粒子a与ssDNA的混合液a中的浓度为10~100 mmol/L;所述的盐在加入了金属纳米粒子a与ssDNA的混合液a中的浓度在6~24 h内增大至最终50~500 mmol/L。The method for preparing a high salt tolerance metal nanoparticle assembly according to claim 1, wherein the coating agent in the step (1) is sodium dodecyl sulfate (SDS); the wrapping agent The mass ratio of the mixture a with the metal nanoparticles a and ssDNA is 0.01 %~1%; the buffer solution is phosphate buffered saline (PBS), pH=7.3, and the concentration of the buffer solution in the mixture a of metal nanoparticles a and ssDNA is 10~100. Mmmol / L; the concentration of the salt in the mixture a of the metal nanoparticles a and ssDNA added increased from 6 to 24 h to the final 50 to 500 mmol / L.
- 根据权利要求1所述的一种高耐盐性金属纳米粒子组装体的制备方法,其特征在于:步骤(1)所述的混合培养的时间为6~24 h;所述的分离纯化的方式为离心分离或者透析。The method for preparing a high salt tolerance metal nanoparticle assembly according to claim 1, wherein the mixing culture time in the step (1) is 6 to 24 h; The separation and purification method is centrifugal separation or dialysis.
- 根据权利要求1所述的一种高耐盐性金属纳米粒子组装体的制备方法,其特征在于:步骤(2)中所述的金属纳米粒子b和步骤(1)中的金属纳米粒子a相同或不同,所述金属纳米粒子b为Au、Ag或Cu中的一种或两种;所述的金属纳米粒子b的尺寸是1~100 nm;所述的金属纳米粒子b与sscDNA的摩尔比为1:0.2~20。The method for preparing a high salt tolerance metal nanoparticle assembly according to claim 1, wherein the metal nanoparticles b in the step (2) are the same as the metal nanoparticles a in the step (1) Or different, the metal nanoparticles b are one or two of Au, Ag or Cu; the size of the metal nanoparticles b is 1~100 The molar ratio of the metal nanoparticle b to the sscDNA is 1:0.2-20.
- 根据权利要求1所述的一种高耐盐性金属纳米粒子组装体的制备方法,其特征在于:步骤(2)所述的包裹剂为SDS,所述包裹剂与加入了金属纳米粒子b与sscDNA的混合液b的质量比为0.01 %~1 %;所述的缓冲溶液为PBS,pH= 7.3,所述的缓冲溶液在加入了金属纳米粒子b与sscDNA的混合液b中的浓度为10~100 mmol/L;所述盐在加入了金属纳米粒子b与sscDNA的混合液b中的浓度为50~500 mmol/L。The method for preparing a high salt-tolerant metal nanoparticle assembly according to claim 1, wherein the wrapping agent in the step (2) is SDS, and the wrapping agent is added with the metal nanoparticle b and The mass ratio of the mixture b of sscDNA is 0.01 %~1%; the buffer solution is PBS, pH=7.3, and the buffer solution has a concentration of 10~100 in the mixture b of the metal nanoparticles b and sscDNA. Mmmol/L; the salt has a concentration of 50 to 500 mmol/L in a mixture b of metal nanoparticles b and sscDNA.
- 根据权利要求1所述的一种高耐盐性金属纳米粒子组装体的制备方法,其特征在于:步骤(2)所述的混合培养的时间为6~24 h;所述的分离纯化方式为离心分离或者透析。The method for preparing a high salt tolerance metal nanoparticle assembly according to claim 1, wherein the mixing culture time in the step (2) is 6 to 24 h; The separation and purification method is centrifugal separation or dialysis.
- 根据权利要求1所述的一种高耐盐性金属纳米粒子组装体的制备方法,其特征在于:步骤(3)所述的产物a和产物b的摩尔比为1:10~400;所述的包裹剂为SDS;所述包裹剂与混合有产物a和产物b的混合溶液的质量比为0.01%~1 %;所述的缓冲溶液为PBS,pH= 7.3;所述缓冲溶液在混合有产物a和产物b的混合溶液中的浓度为10~100 mmol/L;所述的盐在混合有产物a和产物b的混合溶液中的浓度为50~500 mmol/L;所述的混合培养时间为6~24 h;所述的离心转速为5000~20,000 rpm;离心时间为5~30 min,离心1~3次。The method for preparing a high salt-tolerant metal nanoparticle assembly according to claim 1, wherein the molar ratio of the product a to the product b in the step (3) is 1:10 to 400; The wrapping agent is SDS; the mass ratio of the wrapping agent to the mixed solution of product a and product b is 0.01%~1 %; the buffer solution is PBS, pH = 7.3; the concentration of the buffer solution in the mixed solution of product a and product b is 10 to 100 Mmmol / L; the salt in the mixed solution of product a and product b in a concentration of 50 ~ 500 mmol / L; the mixed culture time is 6 ~ 24 h; the centrifugal speed is 5000~20,000 rpm; the centrifugation time is 5~30 min, and the centrifugation is performed 1~3 times.
- 根据权利要求1至8任一项所述制备方法制备得到高耐盐性金属纳米粒子组装体。The high salt-tolerant metal nanoparticle assembly is prepared according to the preparation method according to any one of claims 1 to 8.
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