CN111203546A - Hexadecamer ferritin-gold nanocluster and preparation method thereof - Google Patents
Hexadecamer ferritin-gold nanocluster and preparation method thereof Download PDFInfo
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- B22F9/00—Making metallic powder or suspensions thereof
- B22F9/16—Making metallic powder or suspensions thereof using chemical processes
- B22F9/18—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds
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- B22F1/00—Metallic powder; Treatment of metallic powder, e.g. to facilitate working or to improve properties
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- 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"
- G01N2021/6432—Quenching
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- G01N2223/00—Investigating materials by wave or particle radiation
- G01N2223/10—Different kinds of radiation or particles
- G01N2223/101—Different kinds of radiation or particles electromagnetic radiation
- G01N2223/1016—X-ray
Abstract
The invention relates to a hexadecamer ferritin-gold nanocluster and a preparation method thereof. The method comprises the following steps: a) separating and purifying the hexadecyl polymer ferritin, preparing a hexadecyl polymer ferritin solution with the concentration of 30 mg/mL-60 mg/mL, and preheating for 3-8 minutes at the temperature of 25-40 ℃; b) preparing a chloroauric acid solution with the concentration of 3-10 mM, and preheating for 3-8 minutes at 25-40 ℃; c) uniformly mixing the hexadecyl-polymer ferritin solution and the chloroauric acid solution, wherein the volume ratio of the hexadecyl-polymer ferritin solution to the chloroauric acid solution is 2: 1-1: 3, and adjusting the pH value of the mixed solution to 10-12; d) standing for reaction for 4-24 h at 25-40 ℃ to obtain the hexadecyl polymer ferritin-gold nanocluster solution. The hexadecyl-polymer ferritin-gold nanocluster prepared by the method has better fluorescence intensity and fluorescence life, and mercury ions have higher selectivity, so that the application range of the protein-gold nanocluster can be greatly expanded.
Description
Technical Field
The invention relates to a hexadecamer ferritin-gold nanocluster and a preparation method thereof, and the prepared nanocluster has good fluorescence intensity, long fluorescence life and high selectivity on mercury ions.
Background
Fluorescent metal nanoclusters are an emerging class of fluorescent materials with a size less than 2nm, which have excellent photostability and sub-nanometer-sized biocompatibility, and are easy to synthesize. By utilizing the characteristics, the fluorescent metal nanoclusters have participated in the fields of fluorescence biosensing, biological imaging and the like, and great development is achieved in the fields. The protein is a basic component of a human body and has multiple functions, and by utilizing the protection effect of the protein, the protein-gold nanocluster compound formed by the protein and the gold nanocluster has better fluorescence stability compared with the gold nanocluster, and the functional characteristics of the protein can also expand the application range of the protein-gold nanocluster. Ferritin is a specific class of iron storage and detoxification proteins, which are widely distributed among animals, plants and bacteria, with the exception of yeast. All ferritin has a shell-like structure and possesses the property of self-assembly by which ferritin can be used to entrap some drugs and poorly water soluble, heat labile nutrients. Compared with natural ferritin, the internal cavity of the hexadecamer ferritin is smaller, which is beneficial to the formation of gold nanoclusters. However, how to prepare gold nanoclusters by utilizing the hexameric ferritin has not been reported. The existing gold nanocluster preparation technology is mainly carried out by adopting a reductive protein solution, and the application of the gold nanocluster in the detection aspect is mainly influenced by the stability and the strength of the gold nanocluster.
Disclosure of Invention
Aiming at the technical problems, the invention aims to provide a hexadecamer ferritin-gold nanocluster and a preparation method thereof, wherein ferritin with a small cavity volume is used as a template, so that the stability of the nanoparticle can be improved; in addition, the ferritin can be applied to the field of biomedical imaging by utilizing the specific receptor of ferritin and the characteristic of emitting fluorescence under 650nm, thereby greatly widening the application range of the ferritin.
In order to achieve the purpose, the invention provides the following technical scheme:
a preparation method of a hexadecamer ferritin-gold nanocluster comprises the following steps:
a) separating and purifying the hexadecyl polymer ferritin, preparing a hexadecyl polymer ferritin solution with the concentration of 30 mg/mL-60 mg/mL, and preheating for 3-8 minutes at the temperature of 25-40 ℃;
b) preparing a chloroauric acid solution with the concentration of 3-10 mM, and preheating for 3-8 minutes at 25-40 ℃;
c) uniformly mixing the hexameric ferritin solution prepared in the step a and the chloroauric acid solution prepared in the step b, wherein the volume ratio of the hexameric ferritin solution to the chloroauric acid solution is 2: 1-1: 3, and adjusting the pH value of the mixed solution to 10-12;
d) standing for reaction for 4-24 h at 25-40 ℃ to obtain the hexadecyl polymer ferritin-gold nanocluster solution.
A hexameric ferritin-gold nanocluster prepared according to the preparation method, wherein the size of nanoparticles of the hexameric ferritin-gold nanocluster is 2 ± 1 nm.
The maximum emission wavelength of the hexadecamer ferritin-gold nanocluster is 650nm and the maximum excitation wavelength of the hexadecamer ferritin-gold nanocluster is 500nm in ultraviolet visible spectrum analysis.
The hexadecamer ferritin-gold nanoclusters produce pink fluorescence in fluorescence spectroscopy.
In X-ray photoelectron spectroscopy analysis, the hexadecamer ferritin-gold nanocluster has two peaks between 80eV and 90eV, and the two peaks correspond to 0-valent gold and + 1-valent gold respectively.
6. The hexameric ferritin-gold nanocluster of claim 2, wherein: the hexadecyl-polymer ferritin-gold nanocluster is applied to mercury ion detection.
Compared with the prior art, the invention has the beneficial effects that:
the hexadecamer ferritin-gold nanocluster prepared by the method has better fluorescence intensity and fluorescence life, and mercury ions have higher selectivity, so that the application range of the protein-gold nanocluster can be greatly expanded.
Drawings
FIG. 1 is a graph showing the quenching of the fluorescence of the hexameric ferritin-gold nanoclusters prepared by the present invention by mercury ions of different concentrations.
Detailed Description
The invention is further illustrated with reference to the following figures and examples.
According to the invention, the gold nanoclusters are embedded by means of the cavity structure of the hexadecyl-polymer ferritin to form an ferritin-gold nanocluster compound, the fluorescence stability of the gold nanoclusters is improved by utilizing the protection effect of the protein, and the application range of the ferritin-gold nanoclusters is expanded.
A preparation method of a hexadecamer ferritin-gold nanocluster comprises the following steps:
a) separating and purifying the hexadecyl-polymer ferritin, preparing a hexadecyl-polymer ferritin solution with the concentration of 30 mg/mL-60 mg/mL, and preheating for 3-8 minutes at 25-40 ℃, preferably for 5 minutes;
b) preparing a chloroauric acid solution with the concentration of 3-10 mM, and preheating for 3-8 minutes, preferably for 5 minutes at 25-40 ℃;
c) uniformly mixing the hexameric ferritin solution prepared in the step a and the chloroauric acid solution prepared in the step b, wherein the volume ratio of the hexameric ferritin solution to the chloroauric acid solution is 2: 1-1: 3, and adjusting the pH value of the mixed solution to 10-12;
d) standing for reaction for 4-24 h at 25-40 ℃ to obtain the hexadecyl polymer ferritin-gold nanocluster.
The maximum emission wavelength of the hexadecamer ferritin-gold nanocluster is 650nm and the maximum excitation wavelength of the hexadecamer ferritin-gold nanocluster is 500nm in ultraviolet visible spectrum analysis.
The hexadecamer ferritin-gold nanoclusters produce pink fluorescence in fluorescence spectroscopy.
The nanoparticles of the hexameric ferritin-gold nanoclusters are about 2nm in size.
In X-ray photoelectron spectroscopy analysis, two peaks appear between 80eV and 90eV of the hexadecamer ferritin-gold nanocluster, and the peaks correspond to 0-valent gold and + 1-valent gold respectively.
The hexadecyl-polymer ferritin-gold nanocluster is applied to mercury ion detection.
Examples
Preparation of the hexadecamer ferritin-gold nanocluster:
a) the hexadecamer ferritin was isolated and purified to prepare a solution of 30mg/mL hexadecamer ferritin.
b) A3 mM chloroauric acid solution was prepared and the pH of the solution was adjusted to neutral.
c) The solution of 200. mu.L of hexameric ferritin was pipetted into a 1.5mL centrifuge tube, 200. mu.L of chloroauric acid solution was added thereto, and the mixture was stirred on a magnetic stirrer for 5min at 60r/min, then the pH was adjusted to 11 with 1M sodium hydroxide, and the mixture was stirred on a magnetic stirrer for 30min at 60 r/min.
d) And finally standing for 4 hours at 37 ℃ to obtain the hexadecyl polymer ferritin-gold nanocluster.
The hexadecyl-polymer ferritin-gold nanocluster is used as a mercury ion probe for inorganic mercury detection.
Preparing 5mM mercury chloride solution, preparing a hexameric ferritin-gold nanocluster solution (50 mu M), adding mercury ions with different concentrations into the hexameric ferritin-gold nanocluster solution, mixing uniformly, standing for 5min, and performing fluorescence spectrum scanning with 500nm as excitation wavelength. The specific result is shown in fig. 1, and it can be seen from the figure that the fluorescence intensity is gradually reduced and the fluorescence of the hexameric ferritin-gold nanocluster is gradually quenched as the concentration of mercury ions is increased. The fluorescence of the solution can be completely quenched by adding mercury ions with the final concentration of 70 mu M into the solution of the hexameric ferritin-gold nanocluster with the concentration of 10 mu M.
Claims (6)
1. A preparation method of a hexadecamer ferritin-gold nanocluster is characterized by comprising the following steps: the preparation method comprises the following steps:
a) separating and purifying the hexadecyl polymer ferritin, preparing a hexadecyl polymer ferritin solution with the concentration of 30 mg/mL-60 mg/mL, and preheating for 3-8 minutes at the temperature of 25-40 ℃;
b) preparing a chloroauric acid solution with the concentration of 3-10 mM, and preheating for 3-8 minutes at 25-40 ℃;
c) uniformly mixing the hexameric ferritin solution prepared in the step a and the chloroauric acid solution prepared in the step b, wherein the volume ratio of the hexameric ferritin solution to the chloroauric acid solution is 2: 1-1: 3, and adjusting the pH value of the mixed solution to 10-12;
d) standing for reaction for 4-24 h at 25-40 ℃ to obtain the hexadecyl polymer ferritin-gold nanocluster solution.
2. A hexameric ferritin-gold nanocluster prepared according to the preparation method of claim 1, wherein: the size of the nanoparticle of the hexameric ferritin-gold nanocluster is 2 +/-1 nm.
3. The hexameric ferritin-gold nanocluster of claim 2, wherein: the maximum emission wavelength of the hexadecamer ferritin-gold nanocluster is 650nm and the maximum excitation wavelength of the hexadecamer ferritin-gold nanocluster is 500nm in ultraviolet visible spectrum analysis.
4. The hexameric ferritin-gold nanocluster of claim 2, wherein: the hexadecamer ferritin-gold nanoclusters produce pink fluorescence in fluorescence spectroscopy.
5. The hexameric ferritin-gold nanocluster of claim 2, wherein: in X-ray photoelectron spectroscopy analysis, the hexadecamer ferritin-gold nanocluster has two peaks between 80eV and 90eV, and the two peaks correspond to 0-valent gold and + 1-valent gold respectively.
6. The hexameric ferritin-gold nanocluster of claim 2, wherein: the hexadecyl-polymer ferritin-gold nanocluster is applied to mercury ion detection.
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2020
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