WO2020147753A1 - 丝胶蛋白包裹的金属纳米簇的制备及荧光探针 - Google Patents

丝胶蛋白包裹的金属纳米簇的制备及荧光探针 Download PDF

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WO2020147753A1
WO2020147753A1 PCT/CN2020/072259 CN2020072259W WO2020147753A1 WO 2020147753 A1 WO2020147753 A1 WO 2020147753A1 CN 2020072259 W CN2020072259 W CN 2020072259W WO 2020147753 A1 WO2020147753 A1 WO 2020147753A1
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sericin
metal
aqueous solution
nanoclusters
coated
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PCT/CN2020/072259
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English (en)
French (fr)
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张克勤
仇卉卉
李青松
莫涯
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南通纺织丝绸产业技术研究院
苏州大学
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Priority claimed from CN201910036615.2A external-priority patent/CN109794618A/zh
Priority claimed from CN201910036022.6A external-priority patent/CN109794616A/zh
Priority claimed from CN201910036611.4A external-priority patent/CN109794617A/zh
Application filed by 南通纺织丝绸产业技术研究院, 苏州大学 filed Critical 南通纺织丝绸产业技术研究院
Publication of WO2020147753A1 publication Critical patent/WO2020147753A1/zh

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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
    • B22F1/10—Metallic powder containing lubricating or binding agents; Metallic powder containing organic material
    • B22F1/102—Metallic powder coated with organic material
    • 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
    • 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
    • B22F9/24—Making metallic powder or suspensions thereof using chemical processes with reduction of metal compounds starting from liquid metal compounds, e.g. solutions
    • 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
    • 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
    • C09K11/02—Use of particular materials as binders, particle coatings or suspension media therefor
    • 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

Definitions

  • the invention relates to the field of nanotechnology, in particular to the preparation of a sericin-coated metal nanocluster and a fluorescent probe.
  • Traditional fluorescent probes are mostly organic dyes, quantum dots, up-conversion nanoparticles and fluorescent proteins, but they all have certain defects and deficiencies.
  • the excitation light wavelength range of organic fluorescent dyes is relatively narrow, and different fluorescent dyes usually need excitation light of multiple wavelengths to excite, which brings a lot of inconvenience to the actual research work; in addition, the light stability of organic dyes is not very good, and it is easy to produce light.
  • the bleaching phenomenon is a phenomenon in which the luminescent substance is decomposed by light excitation and the fluorescence intensity is reduced.
  • quantum dots usually have a large particle size, and most of them have a certain degree of toxicity; and non-specifically bound quantum dots will also limit the surface detection sensitivity and cause false signals.
  • fluorescent metal nanoclusters Compared with traditional fluorescent probes, fluorescent metal nanoclusters have the advantages of dimming, ultra-fine sub-nanometer size, high selectivity, and high biocompatibility. They can be used as alternative materials for optical molecular indicators and are simple to synthesize , The harm to the environment is also small. Therefore, in recent years, metal nanoclusters have attracted the attention of many researchers as a new type of fluorescent probe.
  • Metal Nanoclusters are relatively stable nanostructures composed of a few to hundreds of metal atoms, and their size is close to the Fermi wavelength of electrons, compared with large-diameter metal nanoparticles Has more excellent optical, chemical and physical properties.
  • fluorescent gold nanoclusters AuNCs
  • the particle size is generally about a few nanometers, and it has excellent biocompatibility and superior photoinduced properties.
  • Luminescence and excellent light stability are widely used in various scientific fields. Among them, as an important substitute for traditional phosphors, based on the detection mechanism of "off” and “on”, it can be divided into quenched or enhanced fluorescent gold nanoclusters. At present, gold nanoclusters as fluorescent probes have been successfully applied to detect heavy metal ions.
  • the template method As a commonly used method for the synthesis of gold nanoclusters, the template method has unique advantages. It uses various biomolecules, such as proteins, DNA, polymers, etc., as synthetic ligands or templates, which are wrapped on the surface of the nanoclusters and also have protective agents. Function to stabilize the structure and properties of metal nanoclusters. Therefore, it is very important to choose a suitable template to synthesize gold nanoclusters.
  • Biological macromolecules such as proteins have abundant chelating and functional groups such as amino and carboxyl groups, especially sulfhydryl groups. These groups have special affinity with metal atoms and can be used as excellent templates for the synthesis of metal nanoclusters.
  • the current gold nanocluster fluorescent probes synthesized with protein as a template usually require relatively large protein quality, and the prepared gold nanoclusters are not highly targeted, and many proteins are relatively expensive. Therefore, looking for a convenient and easy to obtain , Low-cost protein templates are of great significance.
  • Heavy metal ions such as mercury ions, platinum ions, and arsenic ions have a fatal effect on the life system, and can damage the human brain, nervous system, endocrine system and even kidneys at very low concentrations. In addition, pollution of such heavy metal ions is quite common in the environment. Therefore, the detection of heavy metal ions such as mercury ions and platinum ions is of great significance.
  • electrochemical methods cold atomic absorption spectrometry, inductively coupled plasma mass spectrometry (ICP-MS) and atomic emission spectrometry.
  • ICP-MS inductively coupled plasma mass spectrometry
  • atomic emission spectrometry atomic emission spectrometry
  • fluorescence spectroscopy As an emerging detection method, fluorescence spectroscopy has the advantages of simple and easy operation, extremely high sensitivity and excellent selectivity, and has become a very popular detection method.
  • the current successful mercury ion or platinum ion fluorescent probes also involve complex synthetic routes and related toxicity issues. Therefore, designing a gentle and environmentally friendly mercury ion or platinum ion fluorescent probe has very important practical significance.
  • fluorescent copper nanoclusters have attracted the attention of many researchers as an ideal fluorescent probe.
  • fluorescent analysis is sensitive, simple to operate, selective, accurate, and convenient. Efficient.
  • fluorescent metal nanoclusters have tunable light, ultra-fine sub-nano size, good light stability, high selectivity and high biocompatibility And other advantages, and the synthesis and modification process is simple, and less harmful to the environment.
  • the research of fluorescent copper nanoclusters in catalysis, sensor analysis and detection, cell labeling and cell imaging has been preliminary researched and applied.
  • the purpose of the present invention is to provide a preparation of sericin-coated metal nanoclusters and fluorescent probes.
  • the present invention adopts a one-step method to prepare gold nanoclusters, silver nanoclusters and copper with excellent fluorescence reaction.
  • Nanoclusters have good selectivity and sensitivity to lead ions (Pb 2+ ), mercury ions (Hg 2+ ), platinum ions (Pt 2+ ), and arsenic ions (As 3+ ), respectively.
  • the first object of the present invention is to provide a sericin-coated metal nano-cluster, including a plurality of nanoparticles, the nano-particles include a sericin shell and metal nanoparticles wrapped in the sericin shell,
  • the particle size of metal nanoclusters is 2-5nm.
  • the molecular weight of sericin is 100kDa-150kDa.
  • the metal nanoparticles are gold nanoparticles
  • the metal nanoclusters wrapped by sericin are gold nanoclusters wrapped by sericin
  • the molar ratio of sericin to metal nanoparticles is 1:(5-15).
  • the gold nanoclusters encapsulated by sericin exhibit cyan fluorescence under 365nm ultraviolet light, and the maximum excitation wavelength and maximum emission wavelength are 380nm and 480nm, respectively.
  • the metal nanoparticles are silver nanoparticles
  • the metal nanoclusters wrapped by sericin are silver nanoclusters wrapped by sericin
  • the silver nanoclusters wrapped by sericin emit green fluorescence under the irradiation of 365nm ultraviolet light, at 380nm Excited under the conditions of excitation wavelength, it shows fluorescence emission at 500nm.
  • the metal nanoparticles are copper nanoparticles
  • the metal nanoclusters wrapped by sericin are copper nanoclusters wrapped by sericin
  • the copper nanoclusters wrapped by sericin emit blue-violet fluorescence under the irradiation of 365nm ultraviolet light. Excitation conditions at an excitation wavelength of 350nm show fluorescence emission at 425nm.
  • the mass ratio of copper to sericin is 1:(31-62).
  • the present invention also provides a fluorescent probe for detecting Pb 2+ , which includes the gold nanoclusters coated with sericin of the present invention.
  • concentration of Pb 2+ is 0.01-10 ppm.
  • the aqueous solution of the above-mentioned sericin-coated gold nanoclusters can be used to detect Pb 2+ , and the concentration of the aqueous solution of the sericin-coated gold nanoclusters is 0.02mg/mL-2mg/mL. Preferably, the concentration of the aqueous solution of the sericin-coated gold nanoclusters is 1 mg/mL-2 mg/mL.
  • the present invention also provides a fluorescent probe for detecting Hg 2+ or Pt 2+ , which comprises the silver nanocluster coated with sericin of the present invention.
  • Mercury ions or platinum ions can cause the aggregation of silver nanoclusters, thereby quenching their fluorescence, so silver nanoclusters can be used as effective chemical sensors for detecting mercury ions or platinum ions.
  • the concentration of Hg 2+ or Pt 2+ is 0.01-10 ppm.
  • the aqueous solution of silver nanoclusters coated with sericin can be used to detect Hg 2+ or Pt 2+ , and the concentration of the aqueous solution of silver nanoclusters coated with sericin is 0.02 mg/mL-1.5 mg/mL. Preferably, the concentration of the aqueous solution of the sericin-coated silver nanoclusters is 0.2 mg/mL-1 mg/mL.
  • the present invention also provides a fluorescent probe for detecting As 3+ , which comprises the copper nanoclusters coated with sericin of the present invention.
  • the concentration of As 3+ is 0.01-10 ppm.
  • the aqueous solution of copper nanoclusters encapsulated by sericin can be used to detect As 3+ , and the concentration of the aqueous solution of copper nanoclusters encapsulated by sericin is 1 mg/mL-4 mg/mL. Preferably, the concentration of the aqueous solution of the sericin-coated copper nanoclusters is 1 mg/mL-2 mg/mL.
  • the present invention also provides a preparation method of sericin-coated metal nanoclusters, including the following steps:
  • the gold nanoparticle precursor is tetrachloroalloy acid (HAuCl 4 ) and/or potassium chloroaurate, and the concentration of the aqueous solution of the gold nanoparticle precursor is 8-12 mM.
  • the molar ratio of sericin to Au 3+ in the metal nanoparticle precursor is 1: (5-15).
  • the aqueous solution of the metal salt is an aqueous solution of silver salt
  • the silver salt may be silver nitrate (AgNO 3 ), silver trifluoroacetate (C 2 AgF 3 O 2 ), etc.
  • the concentration of the aqueous solution of silver salt is 8-12 mM.
  • the mass ratio of silver salt and sericin is 1:(28-59).
  • the aqueous solution of the metal salt is an aqueous solution of copper salt
  • the copper salt is copper sulfate (CuSO 4 ), copper chloride (CuCl 2 ), etc.
  • the concentration of the aqueous solution of copper salt is 8-12 mM.
  • the mass ratio of copper salt and sericin is 1: (31-62).
  • the concentration of the sericin aqueous solution is 40-60 mg/mL.
  • alkali is sodium hydroxide (NaOH), ammonia (NH 3 ⁇ H 2 O), and the like.
  • reaction time is 14-16h.
  • Sericin In the silk processing process, the removed sericin is often treated as waste liquid, and there is little recycling of sericin. Sericin accounts for 20% to 30% of silk. Sericin is composed of a variety of proteins from the outside to the inside, and there are many amino acids with long side chains on the sericin chain, such as arginine, lysine, Glutamic acid, methionine, tryptophan, tyrosine, etc., most of the amino acids have strong polar side groups, such as hydroxyl, carboxyl and amino groups.
  • Nano silver clusters are small in size, high in surface activation energy and easy to agglomerate.
  • Sericin contains functional groups such as amino, sulfhydryl, and carboxyl groups, which can chelate precious metal silver to make it more stable.
  • Sericin is used as a protection (template) to prepare silver nanoclusters, which can make the silver nanoclusters have better stability and can effectively prevent their agglomeration.
  • the present invention has at least the following advantages:
  • the present invention recycles and utilizes sericin that is usually treated as waste liquid in silk processing engineering, and applies it to the preparation of metal nanoclusters, and successfully realizes the recycling of resources, and continues the green synthesis route, and synthesizes sericin in one step.
  • the gold nanocluster with the protein as the template has excellent fluorescence response, and initially realizes the stability of the fluorescence emission color and intensity controllability of the copper nanocluster.
  • the invention uses sericin as a reducing agent and protective agent for the synthesis of fluorescent metal nanoclusters.
  • the synthesized fluorescent metal nanoclusters have rare emission wavelengths in the cyan region, have uniform particle size, good fluorescence effect and low toxicity.
  • the experimental method of the present invention is simple, convenient to operate, low in cost, non-polluting to the environment, and solves the problem of waste of resources of sericin processed as waste liquid; it also solves the problem of insufficient particle size of traditional fluorescent probes and insufficient light stability Good, insufficient sensitivity and complicated process.
  • the gold nanoclusters of the present invention can be used as fluorescent probes for detecting Pb 2+ through the fluorescence quenching mechanism, and have good selectivity and high sensitivity to Pb 2+ .
  • the silver nanocluster of the present invention can be used as a fluorescent probe for detecting Hg 2+ or Pt 2+ , and has good selectivity and high sensitivity to Hg 2+ or Pt 2+ .
  • the copper nanocluster of the present invention can be used as a fluorescent probe for detecting As 3+ through the fluorescence quenching mechanism, and has good selectivity and higher sensitivity to As 3+ .
  • the metal nano-cluster fluorescent probe of the present invention has important significance and broadness in the detection of metal ions, bioimaging, and treatment. Application prospects.
  • Figure 1 is the TEM characterization result of the sericin@gold nanocluster powder prepared in Example 1 of the present invention
  • Example 2 is a test result of the maximum emission wavelength of the sericin@gold nanocluster solution and pure silk fibroin solution prepared in Example 1 of the present invention
  • Fig. 3 is a photo of different concentrations of sericin@ ⁇ Nanocluster solution prepared in Example 1 of the present invention irradiated under a 365nm ultraviolet lamp;
  • FIG. 4 is a test result of fluorescence intensity of sericin@ ⁇ Nanocluster solutions of different concentrations prepared in Example 1 of the present invention
  • Fig. 5 is a test result of the application of sericin@ ⁇ Nanocluster to heavy metal ion detection in Example 2 of the present invention.
  • Fig. 6 is the test results of the maximum emission wavelength of the sericin@silver nanocluster solution and the pure silk fibroin solution prepared in Example 5 of the present invention
  • Fig. 7 is a photo of different concentrations of sericin@silver nano cluster solutions prepared in Example 5 of the present invention irradiated under a 365nm ultraviolet lamp;
  • FIG. 8 is a test result of fluorescence intensity of sericin@silver nano cluster solutions of different concentrations prepared in Example 5 of the present invention.
  • FIG. 9 is a test result of application of sericin@silver nanocluster to heavy metal ion detection in Example 6 of the present invention.
  • Example 10 is a test result of the maximum emission wavelength of the sericin@ ⁇ Nanocluster solution and the pure silk fibroin solution prepared in Example 9 of the present invention.
  • Example 11 is a photo of different concentrations of sericin@ ⁇ Nanocluster solution prepared in Example 9 of the present invention irradiated under a fluorescent lamp and a 365nm ultraviolet lamp;
  • FIG. 12 is a test result of the fluorescence intensity of sericin@ ⁇ Nanocluster solutions of different concentrations prepared in Example 9 of the present invention.
  • FIG. 13 is a test result of application of sericin@Cu nanocluster to heavy metal ion detection in Example 10 of the present invention.
  • the method for synthesizing sericin-coated gold nanoclusters includes the following steps:
  • the RF-5301 fluorescence spectrophotometer was used to determine the maximum excitation wavelength and maximum emission wavelength of the sericin@ ⁇ Nanocluster solution at a concentration of 0.5 mg/mL. Scan the range from 300nm to 800nm, and the results showed that the maximum excitation wavelength and maximum emission wavelength were They are 380nm and 480nm respectively ( Figure 2). Compared with the fluorescence spectrum of pure sericin, it can be seen that the fluorescence effect obtained is produced by the prepared sericin@ ⁇ Nanocluster.
  • the RF-5301 fluorescence spectrophotometer was used to measure the relationship between solution concentration and fluorescence intensity, and the results are shown in Figure 4. The results show that when the solution concentration is 1mg/mL-2mg/mL, the fluorescence intensity is stronger.
  • step (1) prepare a 40mg/mL sericin aqueous solution, 8mM potassium chloroaurate aqueous solution and 1M ammonia aqueous solution.
  • step (2) the reaction is carried out at pH 11 and 50°C for 14 hours.
  • step (1) 60mg/mL sericin (Sericin) aqueous solution and 12mM tetrachloroalloy (HAuCl 4 ) aqueous solution are prepared And 1M ammonia solution.
  • step (2) the reaction was carried out at pH 13, 70°C for 14 hours.
  • the method for synthesizing sericin-coated silver nanoclusters includes the following steps:
  • sericin@silver nanocluster solution After placing the sericin@silver nanocluster solution in a refrigerator at -20°C for 12 hours, it was transferred to a freeze dryer at -80°C and freeze-dried for 48 hours to obtain sericin@silver nano cluster powder.
  • the RF-5301 fluorescence spectrophotometer was used to determine the maximum excitation wavelength and maximum emission wavelength of the sericin@ ⁇ Nanocluster solution with a concentration of 1 mg/mL. Scan the range from 300nm to 800nm. The results showed that the maximum excitation wavelength and maximum emission wavelength were respectively It is 380nm and 500nm ( Figure 6). Compared with the fluorescence spectrum of pure sericin, the fluorescence effect obtained is produced by the prepared sericin@ ⁇ Nano Cluster.
  • the RF-5301 fluorescence spectrophotometer was used to measure the relationship between solution concentration and fluorescence intensity, and the results are shown in Figure 8. The results show that when the solution concentration is 0.2mg/mL-1mg/mL, the fluorescence intensity is stronger.
  • the sericin@silver nano clusters were prepared according to the method of Example 1, the difference is:
  • step (1) 40 mg/mL sericin aqueous solution, 8 mM silver trifluoroacetate aqueous solution and 1 M ammonia aqueous solution are prepared.
  • step (2) the reaction is carried out at pH 11 and 50°C for 14 hours.
  • the sericin@silver nano clusters were prepared according to the method of Example 1, except that: in step (1), 60 mg/mL sericin (Sericin) aqueous solution, 12 mM silver trifluoroacetate aqueous solution and 1M ammonia were prepared. Aqueous solution.
  • step (2) the reaction was carried out at pH 13, 70°C for 14 hours.
  • the method for synthesizing a sericin-coated copper nanocluster includes the following steps:
  • the RF-5301 fluorescence spectrophotometer was used to determine the maximum excitation wavelength and maximum emission wavelength of the 4mg/mL sericin@ ⁇ Nanocluster solution, scanning in the range of 300nm to 800nm, the results showed that the maximum excitation wavelength and maximum emission wavelength were respectively It is 350nm and 425nm (Figure 10). Compared with the fluorescence spectrum of pure sericin, it can be seen that the fluorescence effect is produced by the prepared sericin@ ⁇ Nano Cluster.
  • the RF-5301 fluorescence spectrophotometer was used to measure the relationship between solution concentration and fluorescence intensity, and the results are shown in Figure 12. The results show that when the solution concentration is 1mg/mL-2mg/mL, the fluorescence intensity is stronger.
  • step (1) 40mg/mL sericin aqueous solution, 8mM copper chloride aqueous solution and 1M ammonia aqueous solution are prepared.
  • step (2) the reaction is carried out at pH 11 and 50°C for 14 hours.
  • the sericin@Cu nanoclusters were prepared according to the method of Example 1, except that: in step (1), 60 mg/mL sericin aqueous solution, 12 mM copper sulfate aqueous solution and 1 M ammonia aqueous solution were prepared.
  • step (2) the reaction was carried out at pH 13, 70°C for 14 hours.

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Abstract

一种丝胶蛋白包裹的金属纳米簇的制备方法，包括以下步骤：将金属纳米粒子前驱体的水溶液和丝胶蛋白水溶液混匀，然后向其中滴加碱的水溶液调节溶液的pH值至11-13，避光反应，反应完全后得到丝胶蛋白包裹的金属纳米簇的溶液。以及采用该方法制备的金属纳米簇。该金属纳米簇对不同的金属离子具有良好的选择性和灵敏度。

Description

丝胶蛋白包裹的金属纳米簇的制备及荧光探针 技术领域
本发明涉及纳米技术领域，尤其涉及一种丝胶蛋白包裹的金属纳米簇的制备及荧光探针。
背景技术
传统的荧光探针大多为有机染料、量子点、上转换纳米粒子和荧光蛋白等，但都存在一定的缺陷和不足。例如有机荧光染料的激发光波长范围较窄，不同荧光染料通常需要多种波长的激发光来激发，给实际的研究工作带来了很多不便；另外有机染料光稳定性不太好，容易产生光致漂白现象，即由光激发引起发光物质分解而使荧光强度降低的现象。而量子点通常粒径较大，且大部分有一定的毒性；并且非特异性结合的量子点还会限制表面检测灵敏度，导致错误的信号。这些缺陷都限制了它们作为荧光探针的应用。
与传统的荧光探针相比，荧光金属纳米团簇具有可调光性、超细亚纳米尺寸、高选择性和高生物相容性等优点，可作为光学分子指标的替代材料，并且合成简单、对环境的危害也较小。所以近年来，金属纳米簇作为一种新型的荧光探针吸引了很多研究者的关注。
金属纳米簇(Metal Nanoclusters)是一类由几个到数百个金属原子构成的相对稳定的纳米结构，其尺寸接近于电子的费米(Fermi)波长，与大粒径的金属纳米颗粒相比具有更为优异的光学、化学和物理特性。其中，荧光金纳米簇(AuNCs)是金属纳米簇中稳定性最好的纳米材料之一，相关研究开展得最早，粒径一般在几纳米左右，具备优良的生物相容性、优越的光致发光以及优异的光稳定性，被广泛应用在各个科学领域。其中作为传统荧光体的重要替代品，其基于“关闭”和“打开”的检测机制，可以分为淬灭或增强荧光金纳米簇。目前，金纳米簇作为荧光探针已经成功应用于检测重金属离子。
作为金纳米簇的常用合成方法，模板法独具优势，其利用各种生物分子，如蛋白质、DNA、聚合物等作为合成配体也就是模板，包裹在纳米簇表面，同时兼具保护剂的作用以稳定金属纳米簇的结构和性质。因此，选择合适的模板合成金纳米簇非常重要。
生物大分子如蛋白质，具有丰富的螯合和功能基团，如氨基和羧基，尤其是巯基。这些基团与金属原子之间有特殊的亲和力，可以作为合成金属纳米簇的优良模板。但是，目前以蛋白质为模版合成的金纳米簇荧光探针通常需要的蛋白质量比较大，且制备得到的金纳米簇靶向性不强，而很多蛋白质价格比较昂贵，因此寻找一种方便易得、成本低廉的蛋白质模版具有十分重要的意义。
重金属离子例如汞离子、铂离子和砷离子等对生命系统具有致命的影响，在非常低的浓度下也会损害人体的大脑，神经系统，内分泌系统甚至肾脏。并且这类重金属离子在环境中污染相当普遍，因此，汞离子和铂离子等重金属离子的检测具有十分重要的意义。目前，较为成熟的检测汞离子的方法和手段已经有：电化学方法，冷原子吸收光谱法，电感耦合等离子体质谱(ICP-MS)法和原子发射光谱分析法等。但是，这些检测方法具有检测操作繁琐，检测设备价格高昂等不足。而荧光光谱法作为一种新兴的检测方法，具有简单易行的操作方法，极高的灵敏度和优异的选择性等优点，成为一种十分受欢迎的检测手段。不过，目前成功的汞离子或铂离子荧光探针还涉及合成路线复杂以及相关毒性问题。因此，设计一种温和环保的汞离子或铂离子荧光探针具有十分重要的现实意义。
近年来，荧光铜纳米簇作为一种理想的荧光探针吸引了很多研究者的关注，在环境监测和食品安全等领域，荧光分析法分析灵敏、操作简单、选择性好、准确度高且便捷高效。与传统的荧光探针如有机染料、量子点和荧光蛋白等相比，荧光金属纳米团簇具有可调光性、超细亚纳米尺寸、光稳定性好、高选择性和高生物相容性等优点，并且合成与修饰过程简单、对环境的危害也较小。目前，荧光铜纳米簇在催化、传感分析检测、细胞标记和细胞成像等方面的研究已经得到了初步研究和应用。
相比于荧光金纳米簇和银纳米作为新型发光材料的广泛研究和应用，铜纳米簇的合成仍处于初步阶段，其荧光性能也还较少得到实际应用。其限制原因主要在于超小尺寸的铜纳米簇的合成具有一定的难度，且铜离子在空气中容易氧化、聚集。此外，便捷可控、批量生产的方法也一直在探索中。而且，实现铜纳米簇荧光发射颜色及强度可调控性也需要继续深入探究。
发明内容
为解决上述技术问题，本发明的目的是提供一种丝胶蛋白包裹的金属纳米簇的制备及荧光探针，本发明采用一步法制备了具有优异荧光反应的金纳米簇、银纳米簇和铜纳米簇，分别对铅离子(Pb 2+)、汞离子(Hg 2+)或铂离子(Pt 2+)、砷离子(As 3+)具有良好的选择性和灵敏度。
本发明的第一个目的是提供一种丝胶蛋白包裹的金属纳米簇，包括多个纳米粒子，纳米粒子包括丝胶蛋白壳以及包裹在丝胶蛋白壳中的金属纳米粒子，丝胶蛋白包裹的金属纳米簇的粒径为2-5nm。
进一步地，丝胶蛋白的分子量为100kDa-150kDa。
进一步地，金属纳米粒子为金纳米粒子，丝胶蛋白包裹的金属纳米簇为丝胶蛋白包裹的金纳米簇，丝胶蛋白与金属纳米粒子的摩尔比为1:(5-15)。
进一步地，丝胶蛋白包裹的金纳米簇在365nm紫外光的照射下呈现青绿色荧光，其最大激发波长和最大发射波长分别为380nm和480nm。
进一步地，金属纳米粒子为银纳米粒子，丝胶蛋白包裹的金属纳米簇为丝胶蛋白包裹的银纳米簇，丝胶蛋白包裹的银纳米簇在365nm紫外光的照射下发出绿色荧光，在380nm的激发波长条件激发，在500nm处显示荧光发射。
进一步地，金属纳米粒子为铜纳米粒子，丝胶蛋白包裹的金属纳米簇为丝胶蛋白包裹的铜纳米簇，丝胶蛋白包裹的铜纳米簇在365nm紫外光的照射下发出蓝紫色荧光，在350nm的激发波长条件激发，在425nm处显示荧光发射。丝胶蛋白包裹的铜纳米簇中，铜元素和丝胶蛋白的质量比为1:(31-62)。
本发明还提供了一种用于检测Pb 2+的荧光探针，包括本发明上述丝胶蛋白包裹的金纳米簇。
进一步地，Pb 2+的浓度为0.01-10ppm。
进一步地，可将上述丝胶蛋白包裹的金纳米簇的水溶液用于检测Pb 2+，丝胶蛋白包裹的金纳米簇的水溶液的浓度为0.02mg/mL-2mg/mL。优选地，丝胶蛋白包裹的金纳米簇的水溶液的浓度为1mg/mL-2mg/mL。
本发明还提供了一种用于检测Hg 2+或Pt 2+的荧光探针，包括本发明上述丝胶蛋白包裹的银纳米簇。汞离子或铂离子能引起银纳米簇的聚集，从而淬灭其荧光，所以银纳米簇可以作为检测汞离子或铂离子的有效化学传感器。
进一步地，Hg 2+或Pt 2+的浓度为0.01-10ppm。
进一步地，可将上述丝胶蛋白包裹的银纳米簇的水溶液用于检测Hg 2+或Pt 2+，丝胶蛋白包裹的银纳米簇的水溶液的浓度为0.02mg/mL–1.5mg/mL。优选地，丝胶蛋白包裹的银纳米簇的水溶液的浓度为0.2mg/mL-1mg/mL。
本发明还提供了一种用于检测As 3+的荧光探针，包括本发明上述丝胶蛋白包裹的铜纳米簇。
进一步地，As 3+的浓度为0.01-10ppm。
进一步地，可将上述丝胶蛋白包裹的铜纳米簇的水溶液用于检测As 3+，丝胶蛋白包裹的铜纳米簇的水溶液的浓度为1mg/mL-4mg/mL。优选地，丝胶蛋白包裹的铜纳米簇的水溶液的浓度为1mg/mL-2mg/mL。
本发明还提供了一种丝胶蛋白包裹的金属纳米簇的制备方法，包括以下步骤：
将金属纳米粒子前驱体或金属盐的水溶液和丝胶蛋白水溶液混匀，然后向其中滴加碱的水溶液调节溶液的pH值至11-13，并在37℃-70℃下避光反应，反应完全后得到丝胶蛋白包裹的金属纳米簇的溶液。
进一步地，金纳米粒子前驱体为四氯合金酸(HAuCl 4)和/或氯金酸钾，金纳米粒子前驱体的水溶液的浓度为8-12mM。
进一步地，丝胶蛋白与金属纳米粒子前驱体中Au 3+的摩尔比为1:(5-15)。
进一步地，金属盐的水溶液为银盐的水溶液，银盐可为硝酸银(AgNO 3)、三氟乙酸银(C 2AgF 3O 2)等，银盐的水溶液的浓度为8-12mM。
进一步地，银盐和丝胶蛋白的质量比为1:(28-59)。
进一步地，金属盐的水溶液为铜盐的水溶液，铜盐为硫酸铜(CuSO 4)、氯化铜(CuCl 2)等，铜盐的水溶液的浓度为8-12mM。
进一步地，铜盐和丝胶蛋白的质量比为1:(31-62)。
进一步地，丝胶蛋白水溶液的浓度为40-60mg/mL。
进一步地，碱为氢氧化钠(NaOH)、氨水(NH 3·H 2O)等。
进一步地，反应时间为14-16h。
在蚕丝加工过程中，脱去的丝胶蛋白往往当作废液处理，对于丝胶蛋白的回收利用很少。丝胶蛋白占蚕丝量的20％～30％，丝胶蛋白由外到内由多种蛋白质组成，且丝胶蛋白链上有许多侧链较长的氨基酸，如精氨酸、赖氨酸、谷氨酸、甲硫氨酸、色氨酸、酪氨酸等，其中大多数氨基酸具有强极性的侧基，如羟基、羧基和氨基等。
纳米银簇尺寸小、表面活化能高且易团聚，而丝胶蛋白含有氨基、巯基、羧基等官能团，可螯合贵金属银使其更加稳定。丝胶蛋白作为保护(模板)用于制备银纳米簇，可以使银纳米簇具有更好的稳定性，可有效防止其发生团聚。
借由上述方案，本发明至少具有以下优点：
本发明回收利用了通常蚕丝加工工程中作为废液处理的丝胶蛋白，将其应用到金属纳米簇的制备中，成功实现了资源的回收利用，并且延续绿色合成路线，一步法合成以丝胶蛋白为模版的金纳米簇，其具有优异的荧光反应，且初步实现了铜纳米簇荧光发射颜色的稳定及强度可调控性。
本发明利用丝胶蛋白为合成荧光金属纳米簇的还原剂和保护剂，合成的荧光金属纳米簇具有少见的青光区的发射波长，且粒径均一、荧光效果好、毒性低。
本发明的实验方法简单，操作方便，成本低廉，对环境无污染，解决了作为废液处理的丝胶蛋白的资源浪费的问题；也解决了传统荧光探针粒径不够小，光稳定性不够好，灵敏度不足，工艺复杂的问题。
此外，通过荧光淬灭机理本发明的金纳米簇可作为检测Pb 2+的荧光探针，对Pb 2+具有良好的选择性和较高的灵敏度。通过荧光淬灭机理本发明的银纳米簇可作为检测Hg 2+或Pt 2+的荧光探针，对Hg 2+或Pt 2+具有良好的选择性和较高的灵敏度。通过荧光淬灭机理本发明的铜纳米簇可作为检测As 3+的荧光探针，对As 3+具有良好的选择性和较高的灵敏度。
结合丝胶蛋白优异的生物相容性和荧光纳米金属簇在检测和治疗方面的潜力，本发明的金属纳米簇荧光探针在金属离子的检测，生物成像，治疗领域具有重要的意义和广阔的应用前景。
上述说明仅是本发明技术方案的概述，为了能够更清楚了解本发明的技术手段，并可依照说明书的内容予以实施，以下以本发明的较佳实施例并配合附图详细说明如后。
附图说明
图1是本发明实施例1制备的丝胶@金纳米簇粉末的TEM表征结果；
图2是本发明实施例1制备的丝胶@金纳米簇溶液与纯丝素蛋白溶液的最大发射波长测试结果；
图3是本发明实施例1制备的不同浓度的丝胶@金纳米簇溶液在365nm的紫外灯下照射发光的照片；
图4是本发明实施例1制备的不同浓度的丝胶@金纳米簇溶液的荧光强度测试结果；
图5是本发明实施例2中丝胶@金纳米簇应用于重金属离子检测的测试结果；
图6是本发明实施例5制备的丝胶@银纳米簇溶液与纯丝素蛋白溶液的最大发射波长测试结果；
图7是本发明实施例5制备的不同浓度的丝胶@银纳米簇溶液在365nm的紫外灯下照射发光的照片；
图8是本发明实施例5制备的不同浓度的丝胶@银纳米簇溶液的荧光强度测试结果；
图9是本发明实施例6中丝胶@银纳米簇应用于重金属离子检测的测试结果；
图10是本发明实施例9制备的丝胶@铜纳米簇溶液与纯丝素蛋白溶液的最大发射波长测试结果；
图11是本发明实施例9制备的不同浓度的丝胶@铜纳米簇溶液在日光灯和365nm的紫外灯下照射发光的照片；
图12是本发明实施例9制备的不同浓度的丝胶@铜纳米簇溶液的荧光强度测试结果；
图13是本发明实施例10中丝胶@铜纳米簇应用于重金属离子检测的测试结果。
具体实施方式
下面结合附图和实施例，对本发明的具体实施方式作进一步详细描述。以下实施例用于说明本发明，但不用来限制本发明的范围。
实施例1
本发明的一种丝胶蛋白包裹的金纳米簇(以下简称丝胶@金纳米簇)的合成方法，包括以下步骤：
(1)配制50mg/mL的丝胶蛋白(Sericin)水溶液(100kDa-150kDa)，10mM的四氯合金酸(HAuCl 4)水溶液和1M的氢氧化钠(NaOH)水溶液。
(2)将所配的丝胶蛋白水溶液、HAuCl 4水溶液和NaOH水溶液置于37℃的恒温箱中20分钟。将预热后的HAuCl 4水溶液加入丝胶蛋白水溶液中，以1:4的体积比充分混合均匀后，逐滴向混合溶液中加入NaOH水溶液，以调节溶液pH值为12，待三种溶液充分混合均匀后，在37℃的避光恒温箱中充分反应16小时，得到丝胶@金纳米簇的溶液。
将丝胶@金纳米簇溶液置于-20℃冰箱冷冻12小时后，移置-80℃的冷冻干燥机中冻干48小时以获得丝胶@金纳米簇粉末，置于4℃避光环境下保存。对以上获得的丝胶@金纳米簇粉末作TEM表征，结果如图1所示。
取以上制备的丝胶@金纳米簇粉末溶于水，分别配置浓度为0.02mg/mL、0.2mg/mL、0.5mg/mL、1mg/mL、1.5mg/mL和2mg/mL的丝胶@金纳米簇溶液。
应用RF-5301荧光分光光度计测定浓度为0.5mg/mL的丝胶@金纳米簇溶液的最大激发波长和最大发射波长，在300nm到800nm区间进行扫描，结果表明其最大激发波长和最大发射波长分别为380nm和480nm(图2)。与纯丝胶蛋白的荧光图谱对比可得所得荧光效果是由所制丝胶@金纳米簇产生。
取以上不同浓度的丝胶@金纳米簇的溶液，置于在365nm的紫外灯下照射，结果如图3所示，结果表明本发明的丝胶@金纳米簇的溶液在365nm的紫外灯下呈现强烈的青绿色荧光。图3中，从左到右，溶液浓度依次为0.02mg/mL、0.2mg/mL、0.5mg/mL、1mg/mL、1.5mg/mL和2mg/mL。
应用RF-5301荧光分光光度计测定溶液浓度与荧光强度的关系，结果如图4所示。结果表明，溶液浓度为1mg/mL-2mg/mL时，荧光强度较强。
实施例2
称量14等份实施例1制得的丝胶@金纳米簇粉末0.0075g，其中13份分别溶解于5mL的含有其中一种重金属阳离子(Co 2+,Li +,K +,Sr 2+,Ni 2+,Cd 2+,Na +,Hg 2+,Zn 2+,Cu 2+,Ba 2+,As 3+或Pb 2+)水溶液中，另一份溶于5mL超纯水中作为空白对比样，置于37℃恒温振荡箱中，转速200rpm，恒温振荡30mins，以充分有效反应。取丝胶@金纳米簇与重金属阳离子的反应溶液和对比样，使用RF-5301荧光分光光度计进行荧光强度的测定，结果如图5所示，结果表明Pb 2+对丝胶@金纳米簇具有显著的荧光淬灭作用，该丝胶@金纳米簇对Pb 2+具有单一的选择性和较高的灵敏度。此外，测得在0.01ppm-10ppm的Pb 2+浓度范围内，Pb 2+对丝胶@金纳米簇的荧光呈线性淬灭作用。
实施例3
按照实施例1的方法制备丝胶@金纳米簇，不同之处在于：
在步骤(1)中，配置40mg/mL的丝胶蛋白水溶液，8mM的氯金酸钾水溶液和1M的氨水溶液。
在步骤(2)中，在pH值为11、50℃条件下反应14h。
实施例4
按照实施例1的方法制备丝胶@金纳米簇，不同之处在于：在步骤(1)中，配置60mg/mL的丝胶蛋白(Sericin)水溶液，12mM的四氯合金酸(HAuCl 4)水溶液和1M的氨水溶液。
在步骤(2)中，在pH值为13、70℃条件下反应14h。
实施例5
本发明的一种丝胶蛋白包裹的银纳米簇(以下简称丝胶@银纳米簇)的合成方法，包括以下步骤：
(1)配制50mg/mL的丝胶蛋白(Sericin)水溶液(100kDa-150kDa)，10mM的硝酸银(AgNO 3)水溶液和1M的氢氧化钠(NaOH)水溶液。
(2)将所配的丝胶蛋白水溶液、CuSO 4水溶液和NaOH水溶液置于37℃的恒温箱中20分钟。将预热后的AgNO 3水溶液加入丝胶蛋白水溶液中，以1:4的体积比充分混合均匀后，逐滴向混合溶液中加入NaOH水溶液，以调节溶液pH值为12，待三种溶液充分混合均匀后，在37℃的避光恒温箱中充分反应16小时，得到丝胶@银纳米簇的溶液。
将丝胶@银纳米簇溶液置于-20℃冰箱冷冻12小时后，移置-80℃的冷冻干燥机中冻干48小时以获得丝胶@银纳米簇粉末。
取以上制备的丝胶@银纳米簇粉末溶于水，分别配置浓度为0.02mg/mL、0.1mg/mL、 0.2mg/mL、0.5mg/mL、1mg/mL和1.5mg/mL的丝胶@银纳米簇溶液。
应用RF-5301荧光分光光度计测定浓度为1mg/mL的丝胶@银纳米簇溶液的最大激发波长和最大发射波长，在300nm到800nm区间进行扫描，结果表明其最大激发波长和最大发射波长分别为380nm和500nm(图6)，与纯丝胶蛋白的荧光图谱对比可得所得荧光效果是由所制丝胶@银纳米簇产生。
取以上制备的不同浓度的丝胶@银纳米簇的溶液，置于365nm的紫外灯下照射，结果如图7所示，结果表明本发明的丝胶@银纳米簇的溶液在365nm的紫外灯下呈现强烈的绿色荧光，图7中，从左至右，溶液的浓度依次为0.02mg/mL、0.1mg/mL、0.2mg/mL、0.5mg/mL、1mg/mL和1.5mg/mL。
应用RF-5301荧光分光光度计测定溶液浓度与荧光强度的关系，结果如图8所示。结果表明，溶液浓度为0.2mg/mL-1mg/mL时，荧光强度较强。
实施例6
称量15等份实施例1制得的丝胶@银纳米簇粉末0.005g，其中14份分别溶解于5mL的含有一种重金属阳离子(Co 2+,Li +,K +,Sr 2+,Ni 2+,Cd 2+,Na +,Hg 2+,Zn 2+,Cu 2+,Ba 2+,As 3+,Pt 2+或Pb 2+)水溶液中，另一份溶于5mL超纯水中作为空白对比样，置于37℃恒温振荡箱中，转速200rpm，恒温振荡30mins，以充分有效反应。取丝胶@银纳米簇与重金属阳离子的反应溶液和对比样，使用RF-5301荧光分光光度计进行荧光强度的测定，结果如图9所示，结果表明Pt 2+和Hg 2+对丝胶@银纳米簇均具有显著的荧光淬灭作用，该丝胶@银纳米簇对Pt 2+和Hg 2+具有选择性和较高的灵敏度。
实施例7
按照实施例1的方法制备丝胶@银纳米簇，不同之处在于：
在步骤(1)中，配置40mg/mL的丝胶蛋白水溶液，8mM的三氟乙酸银水溶液和1M的氨水溶液。
在步骤(2)中，在pH值为11、50℃条件下反应14h。
实施例8
按照实施例1的方法制备丝胶@银纳米簇，不同之处在于：在步骤(1)中，配置60mg/mL的丝胶蛋白(Sericin)水溶液，12mM的三氟乙酸银水溶液和1M的氨水溶液。
在步骤(2)中，在pH值为13、70℃条件下反应14h。
实施例9
本发明的一种丝胶蛋白包裹的铜纳米簇(以下简称丝胶@铜纳米簇)的合成方法，包括 以下步骤：
(1)配制50mg/mL的丝胶蛋白(Sericin)水溶液(100kDa-150kDa)，10mM的硫酸铜(CuSO 4)水溶液和1M的氢氧化钠(NaOH)水溶液。
(2)将所配的丝胶蛋白水溶液、CuSO 4水溶液和NaOH水溶液置于37℃的恒温箱中20分钟。将预热后的CuSO 4水溶液加入丝胶蛋白水溶液中，以1:4的体积比充分混合均匀后，逐滴向混合溶液中加入NaOH水溶液，以调节溶液pH值为12，待三种溶液充分混合均匀后，在37℃的避光恒温箱中充分反应16小时，得到丝胶@铜纳米簇的溶液。
将丝胶@铜纳米簇溶液置于-20℃冰箱冷冻12小时后，移置-80℃的冷冻干燥机中冻干48小时以获得丝胶@铜纳米簇粉末，置于4℃避光环境下保存。
取以上制备的丝胶@铜纳米簇粉末溶于水，分别配置浓度为1mg/mL、1.5mg/mL、2mg/mL和4mg/mL的丝胶@铜纳米簇溶液。
应用RF-5301荧光分光光度计测定浓度为4mg/mL的丝胶@铜纳米簇溶液的最大激发波长和最大发射波长，在300nm到800nm区间进行扫描，结果表明其最大激发波长和最大发射波长分别为350nm和425nm(图10)，与纯丝胶蛋白的荧光图谱对比可得所得荧光效果是由所制丝胶@铜纳米簇产生。
取浓度为1mg/mL的丝胶@铜纳米簇的溶液，分别置于日光灯和365nm的紫外灯下照射，结果如图11所示，结果表明本发明的丝胶@铜纳米簇的溶液在日光灯下呈黄绿色(图11a)，在365nm的紫外灯下呈现强烈的蓝紫色荧光(图11b)。
应用RF-5301荧光分光光度计测定溶液浓度与荧光强度的关系，结果如图12所示。结果表明，溶液浓度为1mg/mL-2mg/mL时，荧光强度较强。
实施例10
称量14等份实施例1制得的丝胶@铜纳米簇粉末0.01g，其中13份分别溶解于5mL的含有其中一种重金属阳离子(Co 2+,Li +,K +,Sr 2+,Ni 2+,Cd 2+,Na +,Hg 2+,Zn 2+,Cu 2+,Ba 2+,As 3+或Pb 2+)水溶液中，另一份溶于5mL超纯水中作为空白对比样，置于37℃恒温振荡箱中，转速200rpm，恒温振荡30mins，以充分有效反应。取丝胶@铜纳米簇与重金属阳离子的反应溶液和对比样，使用RF-5301荧光分光光度计进行荧光强度的测定，结果如图13所示，结果表明As 3+对丝胶@铜纳米簇具有显著的荧光淬灭作用，该丝胶@铜纳米簇对As 3+具有单一的选择性和较高的灵敏度。
实施例11
按照实施例1的方法制备丝胶@铜纳米簇，不同之处在于：
在步骤(1)中，配置40mg/mL的丝胶蛋白水溶液，8mM的氯化铜水溶液和1M的氨水溶液。
在步骤(2)中，在pH值为11、50℃条件下反应14h。
实施例12
按照实施例1的方法制备丝胶@铜纳米簇，不同之处在于：在步骤(1)中，配置60mg/mL的丝胶蛋白水溶液，12mM的硫酸铜水溶液和1M的氨水溶液。
在步骤(2)中，在pH值为13、70℃条件下反应14h。
以上仅是本发明的优选实施方式，并不用于限制本发明，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明技术原理的前提下，还可以做出若干改进和变型，这些改进和变型也应视为本发明的保护范围。

Claims (20)

  1. 一种丝胶蛋白包裹的金属纳米簇，其特征在于：包括多个纳米粒子，所述纳米粒子包括丝胶蛋白壳以及包裹在所述丝胶蛋白壳中的金属纳米粒子，所述丝胶蛋白包裹的金属纳米簇的粒径为2-5nm。
  2. 根据权利要求1所述的丝胶蛋白包裹的金属纳米簇，其特征在于：所述金属纳米粒子为金纳米粒子，丝胶蛋白与金属纳米粒子的摩尔比为1:(5-15)。
  3. 根据权利要求1所述的丝胶蛋白包裹的金属纳米簇，其特征在于：所述金属纳米粒子为银纳米粒子，所述丝胶蛋白包裹的金属纳米簇为丝胶蛋白包裹的银纳米簇，所述丝胶蛋白包裹的银纳米簇在365nm紫外光的照射下发出绿色荧光，在380nm的激发波长条件激发，在500nm处显示荧光发射。
  4. 根据权利要求1所述的丝胶蛋白包裹的金属纳米簇，其特征在于：所述金属纳米粒子为铜纳米粒子，所述丝胶蛋白包裹的金属纳米簇为丝胶蛋白包裹的铜纳米簇，所述丝胶蛋白包裹的铜纳米簇在365nm紫外光的照射下发出蓝紫色荧光，在350nm的激发波长条件激发，在425nm处显示荧光发射。
  5. 一种用于检测Pb 2+的荧光探针，其特征在于：包括权利要求2所述的丝胶蛋白包裹的金属纳米簇。
  6. 根据权利要求5所述的用于检测Pb 2+的荧光探针，其特征在于：所述Pb 2+的浓度为0.01-10ppm。
  7. 一种用于检测Hg 2+或Pt 2+的荧光探针，其特征在于：包括权利要求3中的丝胶蛋白包裹的金属纳米簇。
  8. 根据权利要求7所述的用于检测Hg 2+或Pt 2+的荧光探针，其特征在于：所述Hg 2+或Pt 2+的浓度为0.01-10ppm。
  9. 一种用于检测As 3+的荧光探针，其特征在于：包括权利要求4中的丝胶蛋白包裹的金属纳米簇。
  10. 根据权利要求9所述的用于检测As 3+的荧光探针，其特征在于：所述As 3+的浓度为0.01-10ppm。
  11. 一种权利要求1中的丝胶蛋白包裹的金属纳米簇的制备方法，其特征在于，包括以下步骤：
    将金属纳米粒子前驱体或金属盐的水溶液和丝胶蛋白水溶液混匀，然后向其中滴加碱的水溶液调节溶液的pH值至11-13，并在37℃-70℃下避光反应，反应完全后得到所述丝胶蛋白包裹的金属纳米簇的溶液。
  12. 根据权利要求11所述的制备方法，其特征在于：所述金属纳米粒子前驱体为四氯合金酸和/或氯金酸钾，所述金属纳米粒子前驱体的水溶液的浓度为8-12mM。
  13. 根据权利要求12所述的制备方法，其特征在于：所述丝胶蛋白与金属纳米粒子前驱体中Au 3+的摩尔比为1:(5-15)。
  14. 根据权利要求11所述的制备方法，其特征在于：所述金属盐的水溶液为银盐的水溶液，所述银盐为硝酸银和/或三氟乙酸银，所述银盐的水溶液的浓度为8-12mM。
  15. 根据权利要求14所述的制备方法，其特征在于：所述银盐和丝胶蛋白的质量比为1:(28-59)。
  16. 根据权利要求11所述的制备方法，其特征在于：所述金属盐的水溶液为铜盐的水溶液，所述铜盐为硫酸铜和/或氯化铜，所述铜盐的水溶液的浓度为8-12mM。
  17. 根据权利要求16所述的制备方法，其特征在于：所述铜盐和丝胶蛋白的质量比为1:(31-62)。
  18. 根据权利要求11所述的制备方法，其特征在于：所述丝胶蛋白水溶液的浓度为40-60mg/mL。
  19. 根据权利要求11所述的制备方法，其特征在于：所述碱为氢氧化钠和/或氨水。
  20. 根据权利要求11所述的制备方法，其特征在于：反应时间为14-16h。
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