CN105092551B - Nitric oxide production method is detected based on the fluorescence noble-metal nanoclusters that bovine serum albumin is modified - Google Patents
Nitric oxide production method is detected based on the fluorescence noble-metal nanoclusters that bovine serum albumin is modified Download PDFInfo
- Publication number
- CN105092551B CN105092551B CN201510500384.8A CN201510500384A CN105092551B CN 105092551 B CN105092551 B CN 105092551B CN 201510500384 A CN201510500384 A CN 201510500384A CN 105092551 B CN105092551 B CN 105092551B
- Authority
- CN
- China
- Prior art keywords
- serum albumin
- bovine serum
- nitric oxide
- fluorescence
- concentration
- 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.)
- Expired - Fee Related
Links
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 title claims abstract description 150
- 108091003079 Bovine Serum Albumin Proteins 0.000 title claims abstract description 36
- 229940098773 bovine serum albumin Drugs 0.000 title claims abstract description 36
- 229910000510 noble metal Inorganic materials 0.000 title claims abstract description 33
- 238000004519 manufacturing process Methods 0.000 title claims 6
- 239000000243 solution Substances 0.000 claims abstract description 43
- 238000000034 method Methods 0.000 claims abstract description 24
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 21
- 239000012047 saturated solution Substances 0.000 claims abstract description 8
- 239000012266 salt solution Substances 0.000 claims abstract description 5
- 239000007864 aqueous solution Substances 0.000 claims abstract description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 3
- 239000003638 chemical reducing agent Substances 0.000 claims description 6
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 claims description 6
- 239000012279 sodium borohydride Substances 0.000 claims description 5
- 229910000033 sodium borohydride Inorganic materials 0.000 claims description 5
- 239000010970 precious metal Substances 0.000 claims description 3
- 239000000523 sample Substances 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims 2
- 238000012986 modification Methods 0.000 claims 2
- 230000004048 modification Effects 0.000 claims 2
- 229910004042 HAuCl4 Inorganic materials 0.000 claims 1
- KHRNKQIDXYINAU-UHFFFAOYSA-N [B+3].[O-2].[Na+].[O-2] Chemical compound [B+3].[O-2].[Na+].[O-2] KHRNKQIDXYINAU-UHFFFAOYSA-N 0.000 claims 1
- JKQOBWVOAYFWKG-UHFFFAOYSA-N molybdenum trioxide Chemical compound O=[Mo](=O)=O JKQOBWVOAYFWKG-UHFFFAOYSA-N 0.000 claims 1
- 230000003287 optical effect Effects 0.000 claims 1
- 239000012888 bovine serum Substances 0.000 abstract description 21
- 239000007850 fluorescent dye Substances 0.000 abstract description 14
- 238000001514 detection method Methods 0.000 abstract description 6
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 239000012086 standard solution Substances 0.000 abstract description 3
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 6
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 6
- 239000010931 gold Substances 0.000 description 6
- 229910052737 gold Inorganic materials 0.000 description 6
- 229910052709 silver Inorganic materials 0.000 description 6
- 239000004332 silver Substances 0.000 description 6
- 238000001727 in vivo Methods 0.000 description 4
- 101710134784 Agnoprotein Proteins 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 150000003378 silver Chemical class 0.000 description 3
- 230000008859 change Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 231100000053 low toxicity Toxicity 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 231100001083 no cytotoxicity Toxicity 0.000 description 2
- 238000010791 quenching Methods 0.000 description 2
- 230000000171 quenching effect Effects 0.000 description 2
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- UQGFMSUEHSUPRD-UHFFFAOYSA-N disodium;3,7-dioxido-2,4,6,8,9-pentaoxa-1,3,5,7-tetraborabicyclo[3.3.1]nonane Chemical compound [Na+].[Na+].O1B([O-])OB2OB([O-])OB1O2 UQGFMSUEHSUPRD-UHFFFAOYSA-N 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002848 electrochemical method Methods 0.000 description 1
- 238000000804 electron spin resonance spectroscopy Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxide Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- 230000003832 immune regulation Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000007180 physiological regulation Effects 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000005062 synaptic transmission Effects 0.000 description 1
Landscapes
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
本发明涉及基于牛血清蛋白修饰的荧光贵金属纳米簇探测一氧化氮的方法，包括：(1)配置牛血清蛋白水溶液，加入贵金属盐溶液，用氢氧化钠调节至pH为11～12，在避光条件下反应8～12小时，以制备牛血清蛋白修饰的荧光贵金属纳米簇；(2)配置NO饱和溶液，稀释成不同浓度的NO标准溶液，用格里斯试剂测各一氧化氮溶液的浓度；(3)将一定浓度的牛血清蛋白修饰的荧光贵金属纳米簇与不同浓度的一氧化氮溶液反应，测定其在不同NO浓度下的荧光强度，构建NO浓度与荧光强度之间的线性关系，得到基于牛血清蛋白修饰的荧光贵金属纳米簇NO荧光探针。与现有技术相比，本发明方便简单，经济，制备的荧光探针选择性高，灵敏性好，对NO的高效探测有巨大的应用价值。The invention relates to a method for detecting nitric oxide based on fluorescent noble metal nanoclusters modified by bovine serum albumin, comprising: (1) preparing an aqueous solution of bovine serum albumin, adding noble metal salt solution, adjusting the pH to 11-12 with sodium hydroxide, and React under light conditions for 8 to 12 hours to prepare bovine serum albumin-modified fluorescent noble metal nanoclusters; (2) configure NO saturated solution, dilute it into NO standard solutions of different concentrations, and use Griess reagent to measure the concentration of each nitric oxide solution (3) react fluorescent noble metal nanoclusters modified with a certain concentration of bovine serum albumin with nitric oxide solutions of different concentrations, measure its fluorescence intensity at different NO concentrations, and construct a linear relationship between NO concentration and fluorescence intensity, A bovine serum albumin-based fluorescent noble metal nanocluster NO fluorescent probe was obtained. Compared with the prior art, the invention is convenient, simple and economical, and the prepared fluorescent probe has high selectivity and good sensitivity, and has great application value for efficient detection of NO.
Description
技术领域technical field
本发明属于生物材料，荧光探针及纳米医药技术领域，具体涉及一种基于牛血清蛋白修饰的荧光金属纳米簇的一氧化氮荧光探针的制法。The invention belongs to the technical field of biological materials, fluorescent probes and nanomedicine, and in particular relates to a method for preparing a nitric oxide fluorescent probe based on bovine serum albumin-modified fluorescent metal nanoclusters.
背景技术Background technique
一氧化氮(nitric oxide,NO)作为信使分子，在人体中有免疫调节、神经传递、血压生理调控等极为重要的生理作用。一氧化氮是一种气体自由基，具有寿命短，活泼性高，易于和其他物质发生反应的特点。因此，在生物体内灵敏地、有选择性地探测一氧化氮就成为一种具有挑战性的工作，也是近几年的研究热点之一。现有的一氧化氮的探测方法主要有化学发光法、分光光度法、电化学方法、电子顺磁共振波谱法等方法，但是这些方法具有耗时长，线性范围窄，精确度不高等缺点，相对于这些方法，荧光探针法具有灵敏度高、选择性好、操作简单等优点。As a messenger molecule, nitric oxide (NO) has extremely important physiological functions such as immune regulation, neurotransmission, and blood pressure physiological regulation in the human body. Nitric oxide is a gaseous free radical, which has the characteristics of short life, high activity and easy reaction with other substances. Therefore, the sensitive and selective detection of nitric oxide in vivo has become a challenging task, and it is also one of the research hotspots in recent years. Existing nitric oxide detection methods mainly include chemiluminescence, spectrophotometry, electrochemical methods, electron paramagnetic resonance spectroscopy and other methods, but these methods have the disadvantages of long time-consuming, narrow linear range, and low accuracy. Compared with these methods, the fluorescent probe method has the advantages of high sensitivity, good selectivity, and simple operation.
现有的一氧化氮荧光探针主要包括含有金属离子的荧光探针和邻苯二胺类有机荧光探针，尽管这些分子有很好的选择性和灵敏性，但是具有一定的细胞毒性，因此，研究一种具有高灵敏性、选择性、高生物相容性及低细胞毒性等优点的一氧化氮荧光探针成为一项重要课题。Existing nitric oxide fluorescent probes mainly include fluorescent probes containing metal ions and o-phenylenediamine organic fluorescent probes. Although these molecules have good selectivity and sensitivity, they have certain cytotoxicity, so , it has become an important topic to study a nitric oxide fluorescent probe with advantages of high sensitivity, selectivity, high biocompatibility and low cytotoxicity.
荧光贵金属纳米簇具有很好的生物相容性和低细胞毒性等特点，在生物医学等方面有很大的用途。牛血清蛋白有极好的生物相容性且没有细胞毒性，所以牛血清蛋白修饰的荧光贵金属纳米簇在生物标记和成像及医学等方面有很大的应用价值。基于NO可以使牛血清蛋白的结构发生改变，从而使牛血清蛋白修饰的荧光贵金属纳米簇荧光淬灭的机理，就可以设计一种高灵敏性和选择性的高效荧光探针，对生物体内NO的探测有巨大的应用价值。Fluorescent noble metal nanoclusters have the characteristics of good biocompatibility and low cytotoxicity, and have great applications in biomedicine and other fields. Bovine serum albumin has excellent biocompatibility and no cytotoxicity, so fluorescent noble metal nanoclusters modified by bovine serum albumin have great application value in biomarkers, imaging and medicine. Based on the mechanism that NO can change the structure of bovine serum albumin, thereby quenching the fluorescence of bovine serum albumin-modified fluorescent noble metal nanoclusters, a highly sensitive and selective high-efficiency fluorescent probe can be designed to detect NO in vivo. The detection has great application value.
目前为止，未见牛血清蛋白稳定的荧光贵金属纳米簇来探测一氧化氮的相关中国专利的报道。因此，基于牛血清蛋白修饰的贵金属纳米簇的一氧化氮荧光探针，发展生物体内NO的荧光探测，这些研究预示了这种特殊的NO探针是一项有意义的工作，也是当前相关研究中亟待解决的关键技术问题。So far, there is no relevant Chinese patent report on bovine serum albumin-stabilized fluorescent noble metal nanoclusters to detect nitric oxide. Therefore, based on the bovine serum albumin-modified noble metal nanocluster fluorescent probe for nitric oxide, the development of fluorescent detection of NO in vivo indicates that this special NO probe is a meaningful work and is also the current relevant research. Key technical issues that need to be resolved urgently.
发明内容Contents of the invention
本发明的目的就是为了克服上述现有技术存在的缺陷而提供一种简单、经济、高生物相容性、低毒性，在生物医药领域拥有良好应用前景的基于牛血清蛋白修饰的荧光贵金属纳米簇探测一氧化氮的方法。The purpose of the present invention is to provide a simple, economical, high biocompatibility, low toxicity, bovine serum albumin-based fluorescent noble metal nanocluster with good application prospects in the field of biomedicine in order to overcome the defects of the above-mentioned prior art Methods for detecting nitric oxide.
本发明的目的可以通过以下技术方案来实现：一种基于牛血清蛋白修饰的荧光贵金属纳米簇探测一氧化氮的方法，其特征在于，该方法具体包括以下步骤：The purpose of the present invention can be achieved through the following technical solutions: a method for detecting nitric oxide based on fluorescent noble metal nanoclusters modified by bovine serum albumin, characterized in that the method specifically includes the following steps:
(1)配置牛血清蛋白水溶液，加入贵金属盐溶液，用氢氧化钠调节至pH为11～12，在避光条件下反应8～12小时，以制备牛血清蛋白修饰的荧光贵金属纳米簇；(1) Prepare an aqueous bovine serum albumin solution, add a noble metal salt solution, adjust the pH to 11-12 with sodium hydroxide, and react for 8-12 hours under dark conditions to prepare bovine serum albumin-modified fluorescent noble metal nanoclusters;
(2)配置NO饱和溶液，稀释成不同浓度的NO标准溶液，用格里斯试剂测各一氧化氮溶液的浓度；(2) Configure NO saturated solution, dilute it into NO standard solutions of different concentrations, and measure the concentration of each nitric oxide solution with Griess reagent;
(3)将一定浓度的牛血清蛋白修饰的荧光贵金属纳米簇与不同浓度的一氧化氮溶液反应，测定其在不同NO浓度下的荧光强度，构建NO浓度与荧光强度之间的线性关系，进而得到基于牛血清蛋白修饰的荧光贵金属纳米簇NO荧光探针。(3) React a certain concentration of bovine serum albumin-modified fluorescent noble metal nanoclusters with different concentrations of nitric oxide solutions, measure its fluorescence intensity at different NO concentrations, and construct a linear relationship between NO concentration and fluorescence intensity, and then A bovine serum albumin-based fluorescent noble metal nanocluster NO fluorescent probe was obtained.
步骤(1)中所述的贵金属盐溶液为HAuCl4.4H2O或AgNO3，其浓度为5～10Mm；牛血清蛋白水溶液的浓度为10～50mg/mL。The noble metal salt solution described in step (1) is HAuCl 4 .4H 2 O or AgNO 3 , the concentration of which is 5-10 Mm; the concentration of bovine serum albumin aqueous solution is 10-50 mg/mL.
步骤(1)调节pH值为11～12后加入硼氢化钠作为还原剂，硼氧化钠的浓度为0.5～10mM。Step (1) After adjusting the pH value to 11-12, sodium borohydride is added as a reducing agent, and the concentration of sodium boron oxide is 0.5-10 mM.
步骤(2)中所述的NO标准溶液的浓度为0.01～2.1mM。The concentration of the NO standard solution described in step (2) is 0.01-2.1 mM.
步骤(3)中所述的牛血清蛋白修饰的荧光贵金属纳米簇与一氧化氮溶液的反应液中贵金属纳米簇的浓度为0.5～2mg/mL。The concentration of the noble metal nanoclusters in the reaction solution of the bovine serum albumin-modified fluorescent noble metal nanoclusters and the nitric oxide solution in step (3) is 0.5-2 mg/mL.
本发明采用有极好的生物相容性且没有细胞毒性的牛血清蛋白为原料修饰的荧光贵金属纳米簇，在生物标记和成像及医学等方面有很大的应用价值。基于NO可以使牛血清蛋白的结构发生改变，从而使牛血清蛋白修饰的荧光贵金属纳米簇荧光淬灭的机理，设计了一种高灵敏性和选择性的高效荧光探针，对生物体内NO进行探测。The invention adopts bovine serum albumin with excellent biocompatibility and no cytotoxicity as the raw material to modify the fluorescent precious metal nanocluster, and has great application value in the fields of biomarking, imaging, medicine and the like. Based on the mechanism that NO can change the structure of bovine serum albumin, thereby quenching the fluorescence of bovine serum albumin-modified fluorescent noble metal nanoclusters, a highly sensitive and selective high-efficiency fluorescent probe was designed to detect NO in vivo. probing.
与现有技术相比，本发明用简单，经济的方法制备出牛血清蛋白修饰的荧光贵金属纳米簇，测定该体系的荧光强度与NO浓度之间的关系，从而获得基于该体系的一氧化氮荧光探针。与现有的探测一氧化氮的技术相比，本发明方法简单，经济，制备的一氧化氮荧光探针生物相容性好，低毒性，荧光强度高，对于NO的高效探测有巨大的应用价值。Compared with the prior art, the present invention prepares bovine serum albumin-modified fluorescent noble metal nanoclusters with a simple and economical method, and measures the relationship between the fluorescence intensity of the system and the NO concentration, thereby obtaining nitric oxide based on the system. fluorescent probe. Compared with the existing technology for detecting nitric oxide, the method of the present invention is simple and economical, and the prepared fluorescent nitric oxide probe has good biocompatibility, low toxicity and high fluorescence intensity, and has great application for efficient detection of NO value.
具体实施方式detailed description
下面对本发明的实施例作详细说明，本实施例在以本发明技术方案为前提下进行实施，给出了详细的实施方式和具体的操作过程，但本发明的保护范围不限于下述的实施例。The following is a detailed description of the embodiments of the present invention. This embodiment is implemented on the premise of the technical solution of the present invention, and detailed implementation methods and specific operating procedures are provided, but the protection scope of the present invention is not limited to the following implementation example.
实施例1Example 1
基于牛血清蛋白修饰的荧光贵金属纳米簇探测一氧化氮的方法，详细的步骤过程如下：配置50mg/mL的牛血清蛋白溶液，加入10mM的HAuCl4.4H2O溶液，两分钟后加入氢氧化钠溶液调节pH约为12，避光条件下反应12小时，以制备牛血清蛋白修饰的荧光金纳米簇。配置浓度为2.1mM的一氧化氮饱和溶液，分别稀释成浓度为0.08mM,0.18mM,0.26mM,0.32mM,0.45mM,0.85mM的一氧化氮溶液。用浓度都为1mg/mL的牛血清蛋白修饰的金纳米簇溶液与上述不同浓度的一氧化氮溶液反应，然后测其在不同NO浓度下的荧光强度，得到NO浓度与荧光强度之间的线性关系，从而得到基于牛血清蛋白修饰的荧光金纳米簇探测一氧化氮的方法。The method for detecting nitric oxide based on bovine serum albumin-modified fluorescent noble metal nanoclusters, the detailed steps are as follows: prepare 50mg/mL bovine serum albumin solution, add 10mM HAuCl 4 .4H 2 O solution, and add hydroxide after two minutes The sodium solution adjusted the pH to about 12, and reacted for 12 hours under the condition of avoiding light, so as to prepare the bovine serum albumin-modified fluorescent gold nanoclusters. Prepare a saturated solution of nitric oxide with a concentration of 2.1mM, and dilute it into nitric oxide solutions with a concentration of 0.08mM, 0.18mM, 0.26mM, 0.32mM, 0.45mM, and 0.85mM respectively. A gold nanocluster solution modified with bovine serum albumin at a concentration of 1 mg/mL reacted with the nitric oxide solution of different concentrations above, and then measured its fluorescence intensity at different NO concentrations, and obtained the linearity between the NO concentration and the fluorescence intensity relationship, thus obtaining a method for detecting nitric oxide based on bovine serum albumin-modified fluorescent gold nanoclusters.
实施例2Example 2
配置25mg/mL的牛血清蛋白溶液，加入5mM的HAuCl4.4H2O溶液，两分钟后加入氢氧化钠溶液调节PH约为12，避光条件下反应12小时，以制备牛血清蛋白修饰的荧光金纳米簇。配置浓度为2.1mM的一氧化氮饱和溶液，分别稀释成浓度0.09mM,0.21mM,0.25mM,0.31mM,0.43mM,0.81mM的一氧化氮溶液。用浓度都为0.5mg/mL的牛血清蛋白修饰的金纳米簇溶液与上述不同浓度的一氧化氮溶液反应，然后测其在不同NO浓度下得荧光强度，得到NO浓度与荧光强度之间的线性关系，从而得到基于牛血清蛋白修饰的荧光金纳米簇探测一氧化氮的方法。Prepare 25mg/mL bovine serum albumin solution, add 5mM HAuCl 4 .4H 2 O solution, add sodium hydroxide solution after two minutes to adjust the pH to about 12, and react for 12 hours under dark conditions to prepare bovine serum albumin-modified Fluorescent gold nanoclusters. Prepare a saturated solution of nitric oxide with a concentration of 2.1mM, and dilute it into nitric oxide solutions with a concentration of 0.09mM, 0.21mM, 0.25mM, 0.31mM, 0.43mM, and 0.81mM respectively. A gold nanocluster solution modified with bovine serum albumin at a concentration of 0.5 mg/mL was reacted with the above-mentioned nitric oxide solutions of different concentrations, and then the fluorescence intensity obtained under different NO concentrations was measured, and the relationship between the NO concentration and the fluorescence intensity was obtained. Linear relationship, thus obtaining a method for detecting nitric oxide based on bovine serum albumin-modified fluorescent gold nanoclusters.
实施例3Example 3
配置50mg/mL的牛血清蛋白溶液，加入10mM的AgNO3溶液，加入氢氧化钠溶液调节PH约为12，再加入10mM的硼氢化钠溶液做还原剂，避光条件下反应12小时，以制备牛血清蛋白修饰的荧光银纳米簇。配置浓度为2.1mM的一氧化氮饱和溶液，分别稀释成浓度为0.13mM,0.38mM,0.56mM,0.86mM,0.94mM,1.20mM的一氧化氮溶液。用浓度都为1mg/mL的牛血清蛋白修饰的银纳米簇与上述不同浓度的一氧化氮溶液反应，然后测其在不同NO浓度下的荧光强度，得到NO浓度与荧光强度之间的线性关系，从而得到基于牛血清蛋白修饰的银纳米簇探测一氧化氮的方法。Prepare 50mg/mL bovine serum albumin solution, add 10mM AgNO 3 solution, add sodium hydroxide solution to adjust the pH to about 12, then add 10mM sodium borohydride solution as reducing agent, and react for 12 hours under dark conditions to prepare Bovine serum albumin-modified fluorescent silver nanoclusters. Prepare a saturated solution of nitric oxide with a concentration of 2.1mM, and dilute it into nitric oxide solutions with a concentration of 0.13mM, 0.38mM, 0.56mM, 0.86mM, 0.94mM, and 1.20mM respectively. The silver nanoclusters modified with bovine serum albumin at a concentration of 1 mg/mL reacted with the above-mentioned nitric oxide solutions of different concentrations, and then measured the fluorescence intensity at different NO concentrations, and obtained the linear relationship between the NO concentration and the fluorescence intensity , thus obtaining a method for detecting nitric oxide based on bovine serum albumin-modified silver nanoclusters.
实施例4Example 4
配置25mg/mL的牛血清蛋白溶液，加入5mM的AgNO3溶液，加入氢氧化钠溶液调节PH约为12，再加入5mM的硼氢化钠溶液做还原剂，避光条件下反应12小时，以制备牛血清蛋白修饰的荧光银纳米簇。配置浓度为2.1mM的一氧化氮饱和溶液，分别稀释成浓度为0.11mM,0.41mM,0.55mM,0.83mM,0.91mM,1.08mM的一氧化氮溶液。用浓度都为1mg/mL的牛血清蛋白修饰的荧光银纳米簇与上述不同浓度的一氧化氮溶液反应，然后测其在不同NO浓度下的荧光强度，得到NO浓度与荧光强度之间的线性关系，从而得到基于牛血清蛋白修饰的银纳米簇探测一氧化氮的方法。Prepare 25mg/mL bovine serum albumin solution, add 5mM AgNO3 solution, add sodium hydroxide solution to adjust the pH to about 12 , then add 5mM sodium borohydride solution as reducing agent, and react for 12 hours under dark conditions to prepare Bovine serum albumin-modified fluorescent silver nanoclusters. Prepare a saturated solution of nitric oxide with a concentration of 2.1mM, and dilute it into nitric oxide solutions with a concentration of 0.11mM, 0.41mM, 0.55mM, 0.83mM, 0.91mM, and 1.08mM respectively. Fluorescent silver nanoclusters modified with bovine serum albumin at a concentration of 1 mg/mL reacted with the above-mentioned nitric oxide solutions of different concentrations, and then measured its fluorescence intensity at different NO concentrations, and obtained the linearity between the NO concentration and the fluorescence intensity relationship, thus obtaining a method for detecting nitric oxide based on bovine serum albumin-modified silver nanoclusters.
实施例5Example 5
配置10mg/mL的牛血清蛋白溶液，加入8mM的AgNO3溶液，加入氢氧化钠溶液调节pH为11～12，再加入8mM的硼氢化钠溶液做还原剂，避光条件下反应8小时，以制备牛血清蛋白修饰的荧光银纳米簇。配置浓度为2.1mM的一氧化氮饱和溶液，分别稀释成浓度为0.01mM,0.41mM,0.55mM,0.83mM,0.91mM,1.08mM的一氧化氮溶液。用浓度都为2mg/mL的牛血清蛋白修饰的荧光银纳米簇与上述不同浓度的一氧化氮溶液反应，然后测其在不同NO浓度下的荧光强度，得到NO浓度与荧光强度之间的线性关系，从而得到基于牛血清蛋白修饰的银纳米簇探测一氧化氮的方法。Prepare 10mg/mL bovine serum albumin solution, add 8mM AgNO 3 solution, add sodium hydroxide solution to adjust the pH to 11-12, then add 8mM sodium borohydride solution as reducing agent, and react for 8 hours under dark conditions. Preparation of bovine serum albumin-modified fluorescent silver nanoclusters. Prepare a saturated solution of nitric oxide with a concentration of 2.1mM, and dilute it into nitric oxide solutions with a concentration of 0.01mM, 0.41mM, 0.55mM, 0.83mM, 0.91mM, and 1.08mM respectively. Fluorescent silver nanoclusters modified with bovine serum albumin at a concentration of 2 mg/mL reacted with the above-mentioned nitric oxide solutions of different concentrations, and then measured their fluorescence intensities at different NO concentrations to obtain the linearity between the NO concentration and the fluorescence intensity relationship, thus obtaining a method for detecting nitric oxide based on bovine serum albumin-modified silver nanoclusters.
Claims (5)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510500384.8A CN105092551B (en) | 2015-08-14 | 2015-08-14 | Nitric oxide production method is detected based on the fluorescence noble-metal nanoclusters that bovine serum albumin is modified |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510500384.8A CN105092551B (en) | 2015-08-14 | 2015-08-14 | Nitric oxide production method is detected based on the fluorescence noble-metal nanoclusters that bovine serum albumin is modified |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105092551A CN105092551A (en) | 2015-11-25 |
| CN105092551B true CN105092551B (en) | 2017-10-10 |
Family
ID=54573466
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510500384.8A Expired - Fee Related CN105092551B (en) | 2015-08-14 | 2015-08-14 | Nitric oxide production method is detected based on the fluorescence noble-metal nanoclusters that bovine serum albumin is modified |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105092551B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105772740B (en) * | 2016-03-17 | 2018-03-27 | 湖北大学 | The preparation method and applications of gold nanoclusters |
| CN105798322B (en) * | 2016-03-17 | 2018-04-24 | 湖北大学 | The preparation method and applications of silver nanoclusters |
| CN108896750B (en) * | 2018-05-11 | 2021-05-25 | 江苏大学 | Preparation method and application of BSA-Au/Ag NCs/OPD/HRP proportional type fluorescence sensor |
| CN110161002A (en) * | 2019-04-28 | 2019-08-23 | 浙江工业大学 | A kind of ratio fluorescence imaging system and its construction method, application |
| CN111849467B (en) * | 2020-08-11 | 2021-08-24 | 苏州大学 | Infrared II region fluorescent gold nanoclusters and their preparation and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6636652B1 (en) * | 1999-08-02 | 2003-10-21 | The Regents Of The University Of Michigan | Optical sensors for the detection of nitric oxide |
| CN102585417A (en) * | 2012-01-04 | 2012-07-18 | 武汉理工大学 | Quantum dot/polymer composite nitric oxide fluorescent probe and preparation method thereof |
| CN104614355A (en) * | 2015-02-06 | 2015-05-13 | 盐城工学院 | A method for detecting Cu2+ concentration based on bovine serum albumin-functionalized gold nanocluster light-scattering probe |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011022677A1 (en) * | 2009-08-21 | 2011-02-24 | Massachusetts Institute Of Technology | Optical nanosensors comprising photoluminescent nanostructures |
| CN106415246A (en) * | 2013-09-16 | 2017-02-15 | 麻省理工学院 | Near infrared fluorescent single walled carbon nanotubes as tissue localizable biosensors |
-
2015
- 2015-08-14 CN CN201510500384.8A patent/CN105092551B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6636652B1 (en) * | 1999-08-02 | 2003-10-21 | The Regents Of The University Of Michigan | Optical sensors for the detection of nitric oxide |
| CN102585417A (en) * | 2012-01-04 | 2012-07-18 | 武汉理工大学 | Quantum dot/polymer composite nitric oxide fluorescent probe and preparation method thereof |
| CN104614355A (en) * | 2015-02-06 | 2015-05-13 | 盐城工学院 | A method for detecting Cu2+ concentration based on bovine serum albumin-functionalized gold nanocluster light-scattering probe |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105092551A (en) | 2015-11-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105092551B (en) | Nitric oxide production method is detected based on the fluorescence noble-metal nanoclusters that bovine serum albumin is modified | |
| Jo et al. | Homogeneous immunosensor based on luminescence resonance energy transfer for glycated hemoglobin detection using upconversion nanoparticles | |
| Li et al. | An ultrasensitive sandwich-type electrochemical immunosensor based on signal amplification strategy of gold nanoparticles functionalized magnetic multi-walled carbon nanotubes loaded with lead ions | |
| Hemmateenejad et al. | BSA-modified gold nanoclusters for sensing of folic acid | |
| Zhou et al. | Determination of dopamine based on its enhancement of gold-silver nanocluster fluorescence | |
| CN103217406B (en) | Based on halfcystine and the Cu of Au/Ag core/shell quantum dot 2+the method for making of fluorescence probe | |
| CN111665356A (en) | SERS-based virus detection method and device | |
| CN104931478B (en) | The method of the surface enhanced Raman scattering effect super sensitivity detection alpha-fetoprotein based on Nano silver grain tripolymer | |
| Omer et al. | Highly efficient gold nano-flower optical biosensor doped in a sol–gel/PEG matrix for the determination of a calcitonin biomarker in different serum samples | |
| Zhang et al. | A gold-silver bimetallic nanocluster-based fluorescent probe for cysteine detection in milk and apple | |
| Li et al. | Sensitive detection of glucose based on gold nanoparticles assisted silver mirror reaction | |
| Yang et al. | Application of silver nanoparticles decorated with β-cyclodextrin in determination of 6-mercaptopurine by surface-enhanced Raman spectroscopy | |
| CN108659815A (en) | Golden copper nanocluster fluorescence probe and preparation method thereof for copper ion detection | |
| CN106596481A (en) | A method for detecting Pb2+ using boron-nitrogen-doped fluorescent carbon dot probes | |
| Liang et al. | Rapid and sensitive colorimetric detection of dopamine based on the enhanced-oxidase mimicking activity of cerium (IV) | |
| CN105866082A (en) | Detection method for content of adenosine in biological sample | |
| Zheng et al. | Improved sensitivity of gold nanoclusters toward trypsin under synergistic adsorption of CdTe quantum dots | |
| CN104330393B (en) | Method for determining glucose by using gold nano-cluster as fluorescence probe | |
| Akrivi et al. | Gold-activated luminol chemiluminescence for the selective determination of cysteine over homocysteine and glutathione | |
| CN102519951B (en) | Color reagent made from colorimetry nanometer material and used for detecting melamine and application of color reagent | |
| CN105158225B (en) | A kind of method of two-photon fluorescence excitation detection melamine | |
| Xi et al. | A fluorescence turn-on strategy to achieve detection of captopril based on Ag nanoclusters | |
| CN103760149A (en) | Method for detecting propranlolum rapidly and flexibly based on nanogold chemiluminiscence | |
| CN104964967B (en) | A kind of method that aniline in water sample is detected using chemical illuminating reagent | |
| CN104237185B (en) | PH value measurement method based on N-acetyl-L-cysteine-gold nanocluster |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20171010 Termination date: 20200814 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |