CN112143490A - Preparation method and application of red fluorescent gold nanocluster and targeting compound thereof - Google Patents
Preparation method and application of red fluorescent gold nanocluster and targeting compound thereof Download PDFInfo
- Publication number
- CN112143490A CN112143490A CN202010905469.5A CN202010905469A CN112143490A CN 112143490 A CN112143490 A CN 112143490A CN 202010905469 A CN202010905469 A CN 202010905469A CN 112143490 A CN112143490 A CN 112143490A
- Authority
- CN
- China
- Prior art keywords
- gold nanocluster
- cells
- red fluorescent
- gold
- prepared
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/08—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials
- C09K11/58—Luminescent, e.g. electroluminescent, chemiluminescent materials containing inorganic luminescent materials containing copper, silver or gold
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/18—Metal complexes
- C09K2211/188—Metal complexes of other metals not provided for in one of the previous groups
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
- G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Materials Engineering (AREA)
- Optics & Photonics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Inorganic Chemistry (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention provides a red fluorescent gold nanocluster and a preparation method and application of a targeting compound thereof. The prepared gold nanoclusters and hyaluronic acid are self-assembled through physical adsorption, so that a targeting gold nanocluster compound is obtained, and the compound can successfully target and identify tumor cells. The red fluorescent gold nanocluster prepared by the invention has the advantages of good water solubility, stable fluorescence property, excellent biocompatibility and low toxicity, and has wide application prospects in the aspects of biological imaging, biological labeling and tumor identification.
Description
Technical Field
The invention relates to a nano material and a targeting compound, in particular to a red fluorescent gold nanocluster and a preparation method and application of the targeting compound.
Background
In recent years, fluorescent gold nanoclusters (AuNCs) have been spotlighted as promising novel fluorescent nanomaterials due to their unique physicochemical and good optical properties. It is reported in the literature that fluorescent AuNCs are typically composed of several to several hundred Au atoms, with dimensions less than 2nm, comparable to the fermi wavelength of electrons. Unlike large gold nanoparticles (AuNPs), ultra-small fluorescent AuNCs do not exhibit surface plasmon resonance absorption in the visible region, but possess significant fluorescence emission in the Near Infrared (NIR) of the visible region. By designing and controlling the size, AuNCs can have the excellent characteristics of larger Stokes shift, quantum confinement effect, long service life, strong photoluminescence, water solubility, light stability, biocompatibility and the like. Thanks to these excellent properties, fluorescent AuNCs have broad application prospects in biomarker, biosensing, bioimaging, and targeted cancer therapy applications.
To obtain fluorescent AuNCs, researchers explored a number of synthetic approaches. Such as ultrasonic synthesis, seed growth method, micro emulsion method, single layer protection method, phase transfer synthesis, etching method, etc. These processes can be generally classified into bottom-up processes and top-down processes. However, the methods are complicated in preparation process, time-consuming and environmentally-friendly, so that the synthesis of fluorescent AuNCs by using a simpler and more environmentally-friendly method is very meaningful. Meanwhile, the AuNCs are functionalized on the basis of synthesis, and diversified clusters are prepared and have more excellent properties, so that the AuNCs can be widely applied. For example, Qiao et al in 2013 use Ova-protected AuNCs as an imaging part, FA as a targeting ligand and a PNAS homopolymer as a linker to prepare a nano conjugate FA-Ova-AuNCs, which indicates that the nano conjugate FA-Ova-AuNCs is expected to be an imaging agent for potential early tumor diagnosis. In 2016, Wang et al used self-assembly of gold nanoclusters with carborane amino derivatives (GNCs-CB) to bioimage cancer cells and target delivery of such carborane compounds into tumors. The nano fluorescent probes open up a new way for targeted tumor detection. At present, the synthesis process of fluorescent AuNCs is complex, and the reports of functionalized fluorescent AuNCs are few. Therefore, it is important to synthesize fluorescent AuNCs by a simple and green method.
In conclusion, aiming at the current situation that the synthesis process of fluorescent AuNCs is complex and few functional fluorescent AuNCs are reported, the method aims to synthesize the fluorescent AuNCs by a simple method and form a gold nanocluster compound by self-assembling Hyaluronic Acid (HA) and the prepared fluorescent AuNCs, and hopes that the fluorescent AuNCs become a targeting system with multiple advantages of stable synthesis technology, stable fluorescence, good biocompatibility, low in vivo toxicity, successful targeting of tumor cells and the like, and a new strategy is provided for identification and diagnosis of clinical cancers.
Disclosure of Invention
The invention aims to provide a preparation method of a red fluorescent gold nanocluster and a compound thereof, the prepared red fluorescent gold nanocluster can be applied to cell imaging, and the prepared fluorescent gold nanocluster compound can be applied to targeted recognition of tumor cells.
The red fluorescent gold nanocluster provided by the invention is prepared by taking lysozyme and tetrachloroauric acid as raw materials and deionized water as a solvent through a microwave reaction.
The invention provides a preparation method of a red fluorescent gold nanocluster, which comprises the following steps:
(1) adding the lysozyme solution and the tetrachloroauric acid solution into a microwave tube, uniformly mixing and stirring;
(2) transferring the mixed solution obtained in the step (1) into a microwave reactor for microwave reaction;
(3) and (3) dialyzing the product obtained in the step (2) to finally obtain a red fluorescent gold nanocluster solution.
The molar ratio of the lysozyme to the tetrachloroauric acid in the step (1) is 1:3-8, preferably 1: 6.
The microwave reaction in the step (2) is carried out at the temperature of 25-70 ℃, the pH value of 7-13 and the time of 5-50 min.
And (3) dialyzing for 48 hours by using a dialysis bag with the molecular weight cutoff of 8000-14000 Da.
The red fluorescent gold nanocluster prepared by the method can be self-assembled with hyaluronic acid through physical adsorption to synthesize a gold nanocluster compound, so that a fluorescent nano system targeting tumor cells is obtained.
The mass ratio of the gold nanoclusters to the hyaluronic acid is 1:3-7, preferably 1: 5.
The invention has the beneficial effects that:
the invention takes lysozyme with good biocompatibility as a protective agent and a reducing agent, and reacts with tetrachloroauric acid through a microwave method to synthesize the red fluorescent gold nanocluster, and the method is simple and environment-friendly.
The prepared gold nanoclusters and hyaluronic acid are subjected to self-assembly through physical adsorption, so that a targeting gold nanocluster compound is obtained, and the compound can successfully target and identify tumor cells.
The red fluorescent gold nanocluster and the gold nanocluster compound prepared by the method are good in water solubility, stable in fluorescence property, excellent in biocompatibility and low in toxicity, and have wide application prospects in aspects of biological imaging, biological labeling and tumor identification.
Drawings
FIG. 1 is a transmission electron micrograph of red fluorescent gold nanoclusters
FIG. 2 is a transmission electron micrograph of gold nanocluster complex
FIG. 3 is the fluorescence excitation and emission spectrum of red fluorescent gold nanocluster
FIG. 4 is an infrared spectrum of red fluorescent gold nanoclusters
FIG. 5 is an X-ray photoelectron spectroscopy analysis chart of red fluorescent gold nanoclusters
FIG. 6 shows the UV-VIS absorption spectrum of gold nanoclusters and gold nanocluster complexes
FIG. 7 shows fluorescence emission spectra of gold nanoclusters and gold nanocluster complexes
FIG. 8 shows the effect of different concentrations of red fluorescent gold nanoclusters on the activity of HeLa and 3T3 cells cultured in vitro
FIG. 9 shows the effect of gold nanocluster complexes of different concentrations on the activity of HeLa and 3T3 cells cultured in vitro
FIG. 10 is a photograph showing the image of the red fluorescence gold nanocluster
FIG. 11 is an image of a targeted cell of gold nanocluster complex
FIG. 12 is a graph showing the quantitative determination of gold nanocluster complexes uptake by tumor cells and normal cells using flow cytometry
Detailed Description
The following examples further illustrate the invention, but the invention is not limited to these examples.
Example 1
Preparation of red fluorescent gold nanoclusters (AuNCs @ Lzm, AuL):
(1) separately, the (4mM, 107.1. mu.L) tetrachloroauric acid aqueous solution and (7.14X 10)-2mM, 1mL) lysozyme aqueous solution is added into a microwave tube and stirred for 5 min;
(2) transferring the mixed solution obtained in the step (1) into a microwave reactor, and carrying out microwave reaction for 5min at 50 ℃;
(3) dialyzing the product obtained in the step (2) for 48 hours by using a dialysis bag with the molecular weight cutoff of 8000-14000 Da to finally obtain a red fluorescent gold nanocluster solution;
(4) further preparing red fluorescent gold nanocluster powder.
Example 2
And (3) characterizing the morphology of the red fluorescent gold nanocluster:
dispersing a small amount of prepared dry red fluorescent gold nanocluster powder in water, dripping the water onto a copper net, drying, and analyzing the appearance of a sample by using a transmission electron microscope.
The electron micrograph of the prepared red gold nanocluster is shown in fig. 1, from which it can be seen that the red gold nanocluster is spherical and has a lattice spacing of 0.25nm, indicating a value of 0.23nm similar to the (111) crystal plane of face centered cubic (fcc) Au/Ag.
Example 3
Fluorescence spectrum determination of red fluorescent gold nanoclusters:
and (3) adding 1mL of red fluorescent gold nanocluster solution into a fluorescence cup, and measuring the fluorescence spectrogram of the clusters by using a fluorescence instrument.
The fluorescence spectrogram of the prepared red fluorescent gold nanocluster is shown in figure 3, and as can be seen from the figure, the optimal excitation wavelength of the cluster is 380nm, the optimal emission wavelength is 650nm, and the cluster is located in a near infrared region so as to avoid the interference of autofluorescence in cells. The inset is a picture taken with ultraviolet radiation (365 nm).
Example 4
Infrared spectrum characterization of red fluorescent gold nanocluster:
a small amount of dried gold nanocluster was mixed with dried KBr (mass ratio of about 1:100), ground and tabletted, and its infrared spectrum was measured.
The infrared spectrogram of the prepared red fluorescent gold nanocluster is shown in fig. 4, and the infrared spectrogram can see that the cluster surface mainly contains hydroxyl and carbonyl. And is located at 868cm-1The peak of (a) was attributed to the quinone-like unsaturated C ═ C structure, probably due to oxidation of the tyrosine residue in lysozyme to quinone by tetrachloroauric acid, indicating successful preparation of the clusters.
Example 5
XPS characterization of red fluorescent gold nanoclusters:
XPS characterization was performed on the dried powder of the red gold nanocluster solution.
The XPS spectrum of the prepared red fluorescent gold nanocluster is shown in fig. 5, and it can be seen from the figure that the XPS spectrum shows five peaks with bonding energies at 284.8eV, 399.8eV, 531.3eV, 169.15eV and 83.6eV, which are respectively assigned to elements C1S, N1S, O1S, S2p and Au4f, indicating that the prepared gold nanocluster is mainly composed of elements C, N, O, S and Au.
Example 6
Toxicity of the red fluorescent gold nanoclusters prepared in example 1 at various concentrations on tumor cells (HeLa) and normal cells (3T3) was examined.
HeLa and 3T3 cells in logarithmic growth phase were first grown at 5.0X 103The concentration of each well is respectively inoculated in a 96-well plate, after the cells are attached to the wall, old culture solution is removed, red fluorescent gold nanoclusters with different concentrations prepared by 200 mu L of DMEM medium are added to be set as an experimental group (A, (0.1mg/mL), B, (0.2mg/mL), C, (0.4mg/mL), D, (0.8mg/mL) and E, (1.2mg/mL), 6 parallel groups are arranged, untreated HeLa and 3T3 cells are used as a control group in each group, MTT solution (20 mu L/well) is added after 24h of culture, and after 5h of incubation, the absorbance of the experimental group and the control group at 490nm is detected by using a microplate reader, so that the cell survival rate is calculated.
The effect of the red fluorescent gold nanoclusters with different concentrations on the cytotoxicity of HeLa and 3T3 cultured in vitro is shown in figure 8, and the graph shows that the clusters are low in toxicity and good in biocompatibility.
Example 7
Application experiment of red fluorescent gold nanoclusters prepared in example 1 in cell imaging:
HeLa cells grown in logarithmic phase (density: 1.0X 10)5One dish) were inoculated in a 35mm petri dish and placed in an incubator (37 ℃, 5% CO)2) Culturing until the cells are completely attached to the wall, removing the old solution, adding 1.5mL of culture solution containing 1.5mg of clusters, incubating for 24h, removing the old solution, washing with cold PBS for 2-3 times, and observing the cluster and cell labeling conditions under a fluorescence microscope.
The application experiment result of the Red fluorescent gold nanocluster in the aspect of cell imaging is shown in a figure 10 (Bright: cell Bright field photo, Red: gold nanocluster complex labeled cell fluorescence imaging (excitation wavelength is 405nm), Merge: superposition field.) when the cell labeling condition is observed under a fluorescence microscope, the cell can be seen to present Bright Red fluorescence, and the Bright Red fluorescence is mainly located in cytoplasm, so that the Red fluorescent gold nanocluster prepared by the method has good application potential in the aspect of cell imaging.
Example 8
Preparation of gold nanocluster complex (AuNCs @ Lzm @ HA, AuLH):
the support used was the red fluorescent gold nanoclusters prepared in example 1.
Hyaluronic Acid (HA) of formula C is used14H22NNaO11The molecular weight is 3000, 25g by Shandong-Xiya chemical Co.
The red fluorescent gold nanocluster is self-assembled into a gold nanocluster compound through physical adsorption and hyaluronic acid, so that a fluorescent nano system targeting tumor cells is obtained, and the preparation method specifically comprises the following steps:
weighing 1mg of red fluorescent gold nanocluster in a microwave tube, adding 1mL of deionized water, fully dissolving, adding 5mg of hyaluronic acid, placing on a stirrer, reacting overnight for 12h, transferring to a 3500Da dialysis bag, placing in a large beaker containing 500mL of deionized water, and stirring for dialysis for 48 h.
Example 9
The gold nanocluster composite prepared in example 8 is subjected to morphology characterization:
the transmission electron micrograph of the prepared gold nanocluster composite is shown in fig. 2, and it can be seen from the micrograph that the cluster composite is spherical.
Example 10
In order to confirm the successful self-assembly of gold nanoclusters and hyaluronic acid, the ultraviolet absorption spectra of the gold nanocluster and gold nanocluster complex were measured, respectively.
The ultraviolet-visible absorption spectrogram of the gold nanocluster and gold nanocluster composite nano system is shown in fig. 6, and it can be seen from the chart that compared with the gold nanoclusters, the ultraviolet absorption peak of the gold nanocluster composite is subjected to red shift, which proves that hyaluronic acid has been successfully loaded on the cluster surface.
Example 11
To further confirm the successful self-assembly of gold nanoclusters and hyaluronic acid, the fluorescence spectra of the gold nanocluster and gold nanocluster complexes were determined, respectively.
The fluorescence spectrogram of the gold nanocluster and gold nanocluster composite nano system is shown in fig. 7, and it can be seen from the graph that the fluorescence intensity of the gold nanocluster composite is slightly reduced compared with that of the gold nanocluster, which proves that hyaluronic acid has been successfully loaded on the cluster surface.
Example 12
Toxicity of the gold nanocluster complexes prepared in example 8 at different concentrations on tumor cells (HeLa) and normal cells (3T3) was examined.
HeLa and 3T3 cells in logarithmic growth phase were first grown at 5.0X 103The concentration of each well is respectively inoculated in a 96-well plate, after the cells are attached to the wall, old culture solution is removed, gold nanocluster compound nano systems with different concentrations prepared by adding 200 mu L of DMEM medium are set as an experimental group (A, (0.1mg/mL), B, (0.2mg/mL), C, (0.4mg/mL), D, (0.8mg/mL) and E, (1.2mg/mL), 6 parallel groups are set, untreated HeLa and 3T3 cells are used as control groups in each group, MTT solution (20 mu L/well) is added after 24h of culture, and after 5h of incubation, the absorbance of 490nm of the experimental group and the control groups is detected by using a microplate reader, so that the cell survival rate is calculated.
The cytotoxicity of the gold nanocluster complex is shown in fig. 9, and it can be seen from the graph that the cell survival rate still reaches more than 80% even if the concentration of the complex reaches 1.2mg/mL, indicating that the synthesized gold nanocluster complex has excellent biocompatibility.
Example 13
Application experiment of gold nanocluster complex prepared in example 8 in targeted cell imaging:
HeLa cells grown in logarithmic phase (density: 1.0X 10)5One dish) were inoculated in a 35mm petri dish and placed in an incubator (37 ℃, 5% CO)2) Culturing until the cells are attached, discarding old solution, washing the cells with PBS buffer solution, and treating the cells with different materials. Specifically, one group of cells was incubated with the gold nanocluster complex (1mg/ml) for 2 hours, and the other group of cells was pretreated with hyaluronic acid (8mg/ml) for 30 minutes and then incubated with the gold nanocluster complex (1mg/ml) for 2 hours. A further group of cells was incubated with gold nanoclusters (1mg/ml) for 2 hours. Normal (3T3) cells were used as a control group and incubated with gold nanocluster complexes (1mg/ml) for 2 hours. Next, all the culture solution in the cell culture dish was aspirated, the cells were washed with PBS, and the cells were fixed with paraformaldehyde. Finally, the cells were observed by fluorescence microscopy and pictures were taken.
Application of the gold nanocluster complex in targeted cell imaging is shown in FIG. 11 (Bright: cell Bright field photograph, Red: gold nanocluster complex labeled cell fluorescence imaging (excitation wavelength of 405nm), Merge: superposition field.) cell labeling is observed under a fluorescence microscope, Bright Red fluorescence appears in tumor cells (HeLa) only treated by the gold nanocluster complex, and the Red fluorescence is weaker in tumor cells pretreated by hyaluronic acid and then treated by the gold nanocluster complex. The red fluorescence in tumor cells treated with gold nanoclusters was also weak. The gold nanocluster compound can target tumor cells. And the red fluorescence in normal cells (3T3) treated by the gold nanocluster compound is very weak, further indicating that the gold nanocluster compound has the tumor targeting property.
Example 14
Targeting of the gold nanocluster complex prepared in example 8 was examined by flow cytometry:
the targeting property of the gold nanocluster compound is quantitatively analyzed by a flow cytometer. HeLa cells, MCF-7 cells and HepG2 cells overexpressed with hyaluronic acid receptors were used as tumor cell models. In contrast, 3T3 cells and DC cells with low expression of hyaluronic acid receptors were selected as normal cell models. Tumor cells were added at 1X 10 per well5The density of individual cells was seeded into 35mm dishes. After 18 hours of cell growth, the cells were first washed with PBS and then treated with different materials. Specifically, one group of cells was incubated with gold nanocluster complex (1mg/mL) for 2 hours, another group of cells was incubated with gold nanoclusters (1mg/mL) for 2 hours, and the other group of cells was pretreated with hyaluronic acid (8mg/mL) for 30 minutes and then incubated with gold nanocluster complex (1mg/mL) for 2 hours. The experimental treatment of normal (3T3) cells was performed by incubating the cells with gold nanocluster complexes (1mg/mL) for 2 hours. After treatment, sucking out all culture solution in the cell culture dish, washing cells with PBS, then, re-suspending the cells in PBS buffer solution through trypsin hydrolysis without EDTA, and finally, detecting the fluorescence intensity of an emission peak at 650nm through flow cytometry to compare uptake of the cells to the nanoparticles in each experimental group so as to judge the targeting property of the gold nanocluster compound.
The targeting results of the gold nanocluster complex prepared in example 8 by flow cytometry are shown in fig. 12, and the fluorescence intensity of the cells such as HeLa, MCF-7 and HepG2 incubated by the gold nanocluster complex is very strong (fig. 12A), which indicates that a large amount of nanoparticles are absorbed by three tumor cells. However, HeLa, MCF-7 and HepG2 cells were pre-treated with free hyaluronic acid for 30 minutes before incubating them with the gold nanocluster complex. The fluorescence intensity was found to be significantly lower in these three cells than in cells without pretreatment, indicating that free hyaluronic acid inhibits nanoparticle uptake by tumor cells. The calculated inhibition rates are 80.56%, 85.45% and 65.04%, respectively, and it can be seen that the targeting effect of the nanoparticles on MCF-7 is optimal, and HeLa cells are followed by HepG2 cells which are the worst, due to the fact that hyaluronic acid receptors (CD44) expressed on the surfaces of different tumor cells are different, HA on the surface of the gold nanocluster complex is specifically combined with CD44 receptors overexpressed on the surfaces of several tumor cells, and the gold nanocluster complex can target the tumor cells. In addition, for the group of normal cells 3T3 and DC cells (low expression CD44) treated by the gold nanocluster complex (fig. 12A), the intracellular fluorescence intensity is very low, which indicates that the two normal cells take up the nanoparticles very little, which indirectly proves that the nanoparticles have tumor targeting. Fig. 12B is a bar graph of the fluorescence intensity in each treated cell, clearly showing that the uptake of gold nanocluster complexes by the above three tumor cells is significantly greater than that of the above normal cells. Thus, the results described above well demonstrate that the prepared gold nanocluster complex can target tumor cells.
Claims (8)
1. A preparation method of red fluorescent gold nanoclusters is characterized by comprising the following steps:
(1) adding the lysozyme solution and the tetrachloroauric acid solution into a microwave tube, uniformly mixing and stirring;
(2) transferring the mixed solution obtained in the step (1) into a microwave reactor for microwave reaction;
(3) and (3) dialyzing the product obtained in the step (2) to finally obtain a red fluorescent gold nanocluster solution.
2. The method of claim 1, wherein the molar ratio of lysozyme to tetrachloroauric acid in step (1) is 1: 3-8.
3. The method of claim 1, wherein the microwave reaction in step (2) is performed at a temperature of 25-70 ℃, a pH of 7-13, and a time of 5-50 min.
4. The method of claim 1, wherein the dialysis in step (3) is performed for 48 hours by using a dialysis bag with a molecular weight cut-off of 8000-14000 Da.
5. A fluorescent gold nanocluster compound is characterized by being prepared by the following method: the red fluorescent gold nanoclusters prepared by the method of claim 1 and hyaluronic acid are self-assembled through physical adsorption.
6. The fluorescent gold nanocluster complex according to claim 5, wherein the mass ratio of the gold nanoclusters to the hyaluronic acid is 1: 3-7.
7. Use of red fluorescing gold nanoclusters prepared according to the method of any one of claims 1 to 4 for the preparation of a reagent for cell imaging.
8. Use of the fluorescent gold nanocluster complex of claim 5 or 6 for preparing a reagent for targeted recognition of tumor cells.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010905469.5A CN112143490A (en) | 2020-09-01 | 2020-09-01 | Preparation method and application of red fluorescent gold nanocluster and targeting compound thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010905469.5A CN112143490A (en) | 2020-09-01 | 2020-09-01 | Preparation method and application of red fluorescent gold nanocluster and targeting compound thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112143490A true CN112143490A (en) | 2020-12-29 |
Family
ID=73890673
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010905469.5A Pending CN112143490A (en) | 2020-09-01 | 2020-09-01 | Preparation method and application of red fluorescent gold nanocluster and targeting compound thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112143490A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113018262A (en) * | 2021-03-12 | 2021-06-25 | 山东大学 | Poly (polidocanol) foam hardening agent stabilizer and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010016803A1 (en) * | 2008-08-05 | 2010-02-11 | Agency For Science, Technology And Research | Methods, compositions, and articles comprising stabilized gold nanoclusters |
| CN103464780A (en) * | 2013-09-06 | 2013-12-25 | 湖南科技大学 | Preparation method of fluorescence gold nano clusters with stable chicken ovalbumin |
| CN105527267A (en) * | 2016-01-30 | 2016-04-27 | 山西大学 | Red fluorescence gold nanocluster as well as preparation method and application thereof |
| KR20160123415A (en) * | 2015-04-15 | 2016-10-26 | 성균관대학교산학협력단 | Method of producing a tumor specific siRNA delivery system containing gold nano particles |
| CN109091679A (en) * | 2018-09-03 | 2018-12-28 | 国家纳米科学中心 | Gold nano-material, preparation method and application |
| CN109705841A (en) * | 2018-12-24 | 2019-05-03 | 山西大学 | A kind of transferrins is the gold nano cluster and its preparation method and application of template |
-
2020
- 2020-09-01 CN CN202010905469.5A patent/CN112143490A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010016803A1 (en) * | 2008-08-05 | 2010-02-11 | Agency For Science, Technology And Research | Methods, compositions, and articles comprising stabilized gold nanoclusters |
| CN103464780A (en) * | 2013-09-06 | 2013-12-25 | 湖南科技大学 | Preparation method of fluorescence gold nano clusters with stable chicken ovalbumin |
| KR20160123415A (en) * | 2015-04-15 | 2016-10-26 | 성균관대학교산학협력단 | Method of producing a tumor specific siRNA delivery system containing gold nano particles |
| CN105527267A (en) * | 2016-01-30 | 2016-04-27 | 山西大学 | Red fluorescence gold nanocluster as well as preparation method and application thereof |
| CN109091679A (en) * | 2018-09-03 | 2018-12-28 | 国家纳米科学中心 | Gold nano-material, preparation method and application |
| CN109705841A (en) * | 2018-12-24 | 2019-05-03 | 山西大学 | A kind of transferrins is the gold nano cluster and its preparation method and application of template |
Non-Patent Citations (3)
| Title |
|---|
| YANG LIU ET AL.,: "Microwave-assisted one-step rapid synthesis of near-infrared gold nanoclusters for NIRF/CT dual-modal bioimaging", 《JOURNAL OF MATERIALS CHEMISTRY B》 * |
| 孙少平等编: "《高分子材料在纳米给药系统中的应用》", 31 August 2017, 黑龙江大学出版社 * |
| 张普: "银、金纳米团簇的合成及其在生化分析中的应用研究", 《中国博士学位论文全文数据库 工程科技Ⅰ辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113018262A (en) * | 2021-03-12 | 2021-06-25 | 山东大学 | Poly (polidocanol) foam hardening agent stabilizer and application thereof |
| CN113018262B (en) * | 2021-03-12 | 2021-12-07 | 山东大学 | Poly (polidocanol) foam hardening agent stabilizer and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109986090B (en) | Double-ligand gold nanoparticle aqueous solution and preparation method and application thereof | |
| CN112680220B (en) | Preparation method and application of fluorescent sulfur quantum dots | |
| CN111303868B (en) | Near-infrared luminescent polypeptide self-assembled gold nano material and preparation method and application thereof | |
| Mishra et al. | Fabrication of gold nanoparticles on biotin-di-tryptophan scaffold for plausible biomedical applications | |
| CN112773899A (en) | Drug delivery carrier based on biological metal organic framework material and preparation method and application thereof | |
| CN112143490A (en) | Preparation method and application of red fluorescent gold nanocluster and targeting compound thereof | |
| Paul et al. | Hydrothermal synthesis of gelatin quantum dots for high-performance biological imaging applications | |
| Wang et al. | Ultra long-term cellular tracing by a fluorescent AIE bioconjugate with good water solubility over a wide pH range | |
| CN113953522A (en) | Electropositive gold nanocluster and preparation method and application thereof | |
| CN108516964B (en) | Nano material with aggregation-induced emission effect and application thereof | |
| Chandra et al. | Fluorescent glyco-gold nanocluster induced EGFR mediated targeting of cancer cells | |
| CN104147608B (en) | Lithium amide soapstone nano particles modified by polyethylene glycol-folic acid as well as preparation and application of lithium amide soapstone nano particles | |
| CN107234238B (en) | A kind of core-shell structure Au@Co (OH)2The preparation method of nanosphere | |
| Wang et al. | Fabrication of hybridized nanoparticles with aggregation-induced emission characteristics and application for cell imaging | |
| Chen et al. | Cytidine mediated AuAg nanoclusters as bright fluorescent probe for tumor imaging in vivo | |
| CN110898228B (en) | Tumor self-targeting photoisomerization nano-carrier and preparation method thereof | |
| CN113789172B (en) | Composite chiral nano assembly and preparation method and application thereof | |
| CN112899231A (en) | Visual tumor cell detection reagent, kit, preparation method and application thereof | |
| CN111687428B (en) | Amphiphilic polymer mediated gold nanoparticle controllable assembly and preparation and application thereof | |
| CN111420072B (en) | Preparation method of MRI-SERS dual-mode contrast agent | |
| CN111848685B (en) | Preparation method of amphiphilic PN = PS type phosphorus-containing tree crown macromolecule nano micelle and application of drug carrier of amphiphilic PN = PS type phosphorus-containing tree crown macromolecule nano micelle | |
| CN109047791B (en) | Gold nanoparticles and synthesis method thereof | |
| CN109400887B (en) | Preparation method of double-fluorescence labeling nano material | |
| CN110205115B (en) | Switch-type CQDs @ Ag core-shell nano fluorescent probe and preparation method and application thereof | |
| CN108841385B (en) | Preparation method and application of fluorescent platinum nanocluster for optical imaging of blood system suspension cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201229 |
|
| RJ01 | Rejection of invention patent application after publication |