CN104267087B - Electrochemical biosensor as well as preparation method and application thereof - Google Patents
Electrochemical biosensor as well as preparation method and application thereof Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title abstract description 6
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims abstract description 48
- 239000000243 solution Substances 0.000 claims abstract description 36
- 150000002343 gold Chemical class 0.000 claims abstract description 30
- 239000011724 folic acid Substances 0.000 claims abstract description 28
- 229960000304 folic acid Drugs 0.000 claims abstract description 28
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 claims abstract description 26
- 235000019152 folic acid Nutrition 0.000 claims abstract description 26
- 102000006815 folate receptor Human genes 0.000 claims abstract description 24
- 108020005243 folate receptor Proteins 0.000 claims abstract description 24
- 229910052753 mercury Inorganic materials 0.000 claims abstract description 20
- -1 mercury ions Chemical class 0.000 claims abstract description 20
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 15
- 229910052737 gold Inorganic materials 0.000 claims abstract description 15
- 239000010931 gold Substances 0.000 claims abstract description 15
- 238000004506 ultrasonic cleaning Methods 0.000 claims abstract description 15
- 229920000642 polymer Polymers 0.000 claims abstract description 14
- 239000012528 membrane Substances 0.000 claims abstract description 10
- 229920000344 molecularly imprinted polymer Polymers 0.000 claims abstract description 9
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract description 8
- 238000005498 polishing Methods 0.000 claims abstract description 7
- 230000009870 specific binding Effects 0.000 claims abstract description 7
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims abstract description 6
- 239000007853 buffer solution Substances 0.000 claims abstract description 4
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- 239000003814 drug Substances 0.000 claims abstract description 4
- 238000001514 detection method Methods 0.000 claims description 17
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 108010052305 exodeoxyribonuclease III Proteins 0.000 claims description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 230000004044 response Effects 0.000 claims description 6
- 239000000523 sample Substances 0.000 claims description 6
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 5
- 230000000694 effects Effects 0.000 claims description 5
- BQPIGGFYSBELGY-UHFFFAOYSA-N mercury(2+) Chemical compound [Hg+2] BQPIGGFYSBELGY-UHFFFAOYSA-N 0.000 claims description 4
- RWQNBRDOKXIBIV-UHFFFAOYSA-N thymine Chemical compound CC1=CNC(=O)NC1=O RWQNBRDOKXIBIV-UHFFFAOYSA-N 0.000 claims description 4
- 108020004414 DNA Proteins 0.000 claims description 3
- 239000003792 electrolyte Substances 0.000 claims description 3
- 239000000276 potassium ferrocyanide Substances 0.000 claims description 3
- 229920001059 synthetic polymer Polymers 0.000 claims description 3
- XOGGUFAVLNCTRS-UHFFFAOYSA-N tetrapotassium;iron(2+);hexacyanide Chemical compound [K+].[K+].[K+].[K+].[Fe+2].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] XOGGUFAVLNCTRS-UHFFFAOYSA-N 0.000 claims description 3
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 3
- 239000012498 ultrapure water Substances 0.000 claims description 3
- 238000006243 chemical reaction Methods 0.000 claims description 2
- 125000003929 folic acid group Chemical group 0.000 claims description 2
- 229940113082 thymine Drugs 0.000 claims description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims 2
- 229910052742 iron Inorganic materials 0.000 claims 1
- NNFCIKHAZHQZJG-UHFFFAOYSA-N potassium cyanide Chemical compound [K+].N#[C-] NNFCIKHAZHQZJG-UHFFFAOYSA-N 0.000 claims 1
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- 229920005597 polymer membrane Polymers 0.000 abstract 1
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- 238000000034 method Methods 0.000 description 7
- 238000003776 cleavage reaction Methods 0.000 description 6
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- 238000001318 differential pulse voltammogram Methods 0.000 description 4
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- 230000008859 change Effects 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 2
- 238000011160 research Methods 0.000 description 2
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- 238000011953 bioanalysis Methods 0.000 description 1
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Abstract
Description
技术领域technical field
本发明属于生物传感器技术领域，具体涉及用模板分子与聚合物构成的聚合物膜组装电化学传感器，并用此传感器对叶酸受体及汞离子进行分析检测。The invention belongs to the technical field of biosensors, and in particular relates to an electrochemical sensor assembled with a polymer film composed of template molecules and polymers, and uses the sensor to analyze and detect folic acid receptors and mercury ions.
背景技术Background technique
在传感器领域中，分子印迹技术由于其具有很高的选择识别属性、简单的合成及很高的稳定性，而引起研究者们广泛的探究兴趣。今年来电化学生物传感器因为制造成本低、仪器简单、操作简单和相应快速等优点而成为最有前景的分析方法之一。因此结合了这两者固有优势的基于分子印迹聚合物膜的电化学生物传感器，在生物分析、药物分析、环境监测和评估以及农药残留检测等领域具有巨大的潜力，也引起及其强烈的关注和研究。然而目前，大部分已报道的基于分子印迹聚合物膜的电化学生物传感器仅仅用于检测一种分析物——模板分子。很少有关研究体系可检测多种目标分析物。本申请尝试打破这个基于分子印迹聚合物膜的电化学生物传感器的瓶颈。In the field of sensors, molecular imprinting technology has aroused extensive interest of researchers due to its high selective recognition properties, simple synthesis and high stability. Electrochemical biosensors have become one of the most promising analytical methods this year because of their low manufacturing cost, simple instrumentation, simple operation, and rapid response. Therefore, electrochemical biosensors based on molecularly imprinted polymer membranes, which combine the inherent advantages of the two, have great potential in the fields of bioanalysis, drug analysis, environmental monitoring and evaluation, and pesticide residue detection, and have also attracted strong attention. and research. However, most of the reported electrochemical biosensors based on molecularly imprinted polymer membranes are only used to detect one analyte—template molecule. Few of the research systems detect multiple target analytes. This application attempts to break the bottleneck of this electrochemical biosensor based on molecularly imprinted polymer membranes.
发明内容Contents of the invention
本发明的目的在于提供一种电化学生物传感器，及其制备方法和用途，利用目标分析物与分子印迹膜中的模板分子的特异性结合作用，利用汞离子-胸腺嘧啶-汞离子碱基错配作用再通过外切酶III的切割作用释放模板分子，实现对叶酸受体及汞离子进行分析检测。具体技术方案如下：The purpose of the present invention is to provide an electrochemical biosensor, its preparation method and application, using the specific binding effect of the target analyte and the template molecule in the molecularly imprinted membrane, using the mercury ion-thymine-mercury ion base error After complexation, the template molecules are released through the cleavage of exonuclease III, so as to realize the analysis and detection of folic acid receptors and mercury ions. The specific technical scheme is as follows:
一种电化学生物传感器的制备方法，包括如下步骤：A method for preparing an electrochemical biosensor, comprising the steps of:
(1)将合成聚合物膜的药品配成特定浓度的溶液备用；(1) The medicine of synthetic polymer membrane is made into the solution of specific concentration for subsequent use;
(2)将DNA序列溶解在Tris-HCl缓冲溶液中备用；(2) Dissolving the DNA sequence in a Tris-HCl buffer solution for subsequent use;
(3)将金电极进行抛光处理，再进行超声波清洗；(3) gold electrode is carried out polishing treatment, carries out ultrasonic cleaning again;
(4)以处理干净的裸金电极为工作电极，以含有一定浓度模板分子的聚合物溶液为电解液，通过电聚合方法，合成聚合物修饰的金电极；(4) Using a clean bare gold electrode as a working electrode, using a polymer solution containing a certain concentration of template molecules as an electrolyte, and synthesizing a polymer-modified gold electrode by electropolymerization;
(5)将得到的聚合物修饰金电极浸泡清洗，将聚合物中的模板分子洗脱掉，得到分子印迹膜修饰的金电极；(5) soaking and cleaning the obtained polymer-modified gold electrode, eluting the template molecules in the polymer to obtain a molecularly imprinted membrane-modified gold electrode;
(6)得到基于分子印迹聚合物膜构建的电化学生物传感器。(6) Obtain an electrochemical biosensor based on molecularly imprinted polymer membrane.
进一步地，步骤(2)中，DNA序列采用一端氨基化，Tris-HCl的pH值为7.4，并在低温下下保存。Further, in step (2), the DNA sequence is aminated at one end, the pH value of Tris-HCl is 7.4, and stored at low temperature.
进一步地，步骤(3)中，包括如下步骤：Further, in step (3), the following steps are included:
(2-1)将金电极用0.3μm的铝粉进行抛光处理；(2-1) Polishing the gold electrode with 0.3 μm aluminum powder;
(2-2)将金电极用0.5μm的铝粉进行抛光处理；(2-2) Polishing the gold electrode with 0.5 μm aluminum powder;
(2-3)放入HNO3:H2O(v/v)＝1:1溶液，进行超声波清洗，超声清洗的时间为3～5min；(2-3) Put in HNO 3 :H 2 O(v/v)=1:1 solution, and perform ultrasonic cleaning, and the ultrasonic cleaning time is 3-5 minutes;
(2-4)放入乙醇溶液，进行超声波清洗，超声清洗的时间为3～5min；(2-4) put into ethanol solution, and perform ultrasonic cleaning, and the time of ultrasonic cleaning is 3 to 5 minutes;
(2-5)放入超纯水中，进行超声波清洗，超声清洗的时间为3～5min。(2-5) Put it into ultrapure water, and perform ultrasonic cleaning, and the time of ultrasonic cleaning is 3 to 5 minutes.
进一步地，步骤(4)中，模板分子为叶酸。Further, in step (4), the template molecule is folic acid.
进一步地，步骤(5)中，将聚合物修饰金电极置于含甲醇和醋酸的溶液中浸泡清洗约20～30min，用以将聚合物中的叶酸洗脱掉。Further, in step (5), the polymer-modified gold electrode is soaked and cleaned in a solution containing methanol and acetic acid for about 20-30 minutes, so as to elute the folic acid in the polymer.
一种电化学生物传感器，采用上述方法制备得到。An electrochemical biosensor is prepared by the above-mentioned method.
上述电化学生物传感器的用途，用于对叶酸受体进行检测。The above-mentioned electrochemical biosensor is used for detecting folic acid receptors.
进一步地，所述检测方法包括：将不同浓度的叶酸受体加入至含有确定浓度的叶酸溶液中培养约40～60min；待叶酸受体与叶酸结合完全，将制备得到的分子印迹膜修饰的金电极置于叶酸和叶酸受体反应后的溶液中，实现对叶酸受体的间接检测。Further, the detection method includes: adding different concentrations of folic acid receptors into a solution containing folic acid of a certain concentration and incubating for about 40-60 minutes; The electrode is placed in the solution after the folic acid and the folic acid receptor have reacted to realize the indirect detection of the folic acid receptor.
上述电化学生物传感器的用途，用于对汞离子进行检测。The above-mentioned electrochemical biosensor is used for detecting mercury ions.
进一步地，所述检测方法包括如下步骤：将不同浓度的汞离子加入至具有确定浓度的叶酸修饰的探针DNA溶液中培养30～50min，将制备得到的分子印迹膜修饰的金电极放在外切酶III切割后的溶液中，实现对汞离子的间接检测。Further, the detection method includes the following steps: adding different concentrations of mercury ions to the folic acid-modified probe DNA solution with a determined concentration and culturing for 30-50 minutes, and placing the prepared molecularly imprinted membrane-modified gold electrode on the exosome In the solution after enzyme III cleavage, the indirect detection of mercury ions is realized.
与目前现有技术相比，本发明制备的电化学生物传感器，结合了电化学生物传感器和分子印迹技术两者固有的优势，且合成方法简单、耗能低、成本低。利用目标分析物与模板分子——叶酸的特异性结合作用，或通过外切酶III的切割作用释放模板分子，达到分别检测叶酸受体和汞离子的目的。本发明使用电沉积法组装基于分子印迹聚合物膜的电化学生物传感器，利用目标分析物与分子印迹膜中的模板分子的特异性结合作用，利用汞离子-胸腺嘧啶-汞离子碱基错配作用再通过外切酶III的切割作用释放模板分子，从而实现对目标分析物检测的间接检测。此传感器实现了对目标分析物灵敏性、特异性、稳定性的检测。Compared with the current prior art, the electrochemical biosensor prepared by the present invention combines the inherent advantages of both the electrochemical biosensor and the molecular imprinting technology, and has a simple synthesis method, low energy consumption and low cost. The purpose of detecting folate receptor and mercury ion is achieved by using the specific binding effect of the target analyte on the template molecule - folic acid, or releasing the template molecule through the cleavage of exonuclease III. The present invention uses the electrodeposition method to assemble the electrochemical biosensor based on the molecularly imprinted polymer film, utilizes the specific binding effect between the target analyte and the template molecule in the molecularly imprinted film, and utilizes the base mismatch of mercury ion-thymine-mercury ion The template molecule is released through the cleavage of exonuclease III, so as to realize the indirect detection of the target analyte. The sensor realizes the detection of the sensitivity, specificity and stability of the target analyte.
附图说明Description of drawings
图1(a)为叶酸印迹膜修饰的金电极的扫描电子显微镜图片；Fig. 1 (a) is the scanning electron microscope picture of the gold electrode modified by folic acid imprinted membrane;
图1(b)为聚合物修饰的金电极的扫描电子显微镜图片；Figure 1(b) is a scanning electron microscope image of a polymer-modified gold electrode;
图2(a)为制备聚合物修饰金电极的电聚合循环伏安图。Figure 2(a) is the electropolymerization cyclic voltammogram for preparing polymer-modified gold electrodes.
图2(b)为每一步电极修饰过程电极的阻抗图。Figure 2(b) is the impedance diagram of the electrode in each step of the electrode modification process.
图中：In the picture:
a为裸的金电极的阻抗图。a is the impedance map of the bare gold electrode.
b为聚合物修饰金电极的阻抗图。b is the impedance map of the polymer-modified gold electrode.
c为分子印迹膜修饰的金电极的阻抗图。c is the impedance map of the molecularly imprinted membrane-modified gold electrode.
d为分子印迹膜修饰的金电极在一定浓度模板分子溶液中培养后的阻抗图。d is the impedance diagram of the molecularly imprinted membrane-modified gold electrode incubated in a certain concentration of template molecule solution.
图2(c)为不同电极的循环伏安图。Figure 2(c) is the cyclic voltammogram of different electrodes.
图中in the picture
a为聚合物修饰电极的循环伏安图。a is the cyclic voltammogram of the polymer modified electrode.
b为分子印迹膜修饰的金电极的循环伏安图。b is the cyclic voltammogram of the molecularly imprinted membrane-modified gold electrode.
c为分子印迹膜修饰的金电极在一定浓度模板分子溶液中培养后的循环伏安图。c is the cyclic voltammogram of the molecularly imprinted membrane-modified gold electrode incubated in a certain concentration of template molecule solution.
图2(d)不同电极的差示脉冲伏安图。Fig. 2(d) Differential pulse voltammograms of different electrodes.
图中in the picture
a为聚合物修饰电极的差示脉冲伏安图。a is the differential pulse voltammogram of the polymer modified electrode.
b为分子印迹膜修饰的金电极的差示脉冲伏安图。b is the differential pulse voltammogram of the molecularly imprinted membrane-modified gold electrode.
c为分子印迹膜修饰的金电极在一定浓度模板分子溶液中培养后的差示脉冲伏安图。c is the differential pulse voltammogram of the molecularly imprinted membrane-modified gold electrode incubated in a certain concentration of template molecule solution.
图3(a)为基于分子印迹聚合物膜构建电化学生物传感器对不同浓度目标叶酸受体检测的循环伏安图。Fig. 3(a) is the cyclic voltammogram of the detection of different concentrations of target folate receptors by the electrochemical biosensor based on the molecularly imprinted polymer film.
图3(b)为此传感器对不同浓度叶酸受体响应的标准曲线。Figure 3(b) is the standard curve of the sensor's response to different concentrations of folate receptors.
图4(a)为基于分子印迹聚合物膜构建电化学生物传感器对不同浓度目标汞离子检测的循环伏安图。Figure 4(a) is the cyclic voltammogram of the detection of different concentrations of target mercury ions by the electrochemical biosensor based on the molecularly imprinted polymer film.
图4(b)为此传感器对不同浓度汞离子响应的标准曲线。Figure 4(b) is the standard curve of the sensor's response to different concentrations of mercury ions.
图5为基于叶酸印迹聚合物膜的电化学传感器的制备及其对叶酸受体及汞离子的检测的简单原理图。Fig. 5 is a simple schematic diagram of the preparation of the electrochemical sensor based on the folic acid imprinted polymer film and its detection of folic acid receptors and mercury ions.
具体实施方式detailed description
下面根据附图对本发明进行详细描述，其为本发明多种实施方式中的一种优选实施例。The present invention will be described in detail below according to the accompanying drawings, which is a preferred embodiment among various implementations of the present invention.
基于叶酸印迹聚合物膜的电化学传感器的制备及其对叶酸受体及汞离子的检测的步骤如下：The preparation of the electrochemical sensor based on the folic acid imprinted polymer film and the steps of its detection of the folic acid receptor and mercury ions are as follows:
a、将购买的合成聚合物膜的药品配成特定浓度的溶液备用，将购买的DNA序列(一端氨基化)溶解在Tris-HCl(pH7.4)缓冲溶液中，并在低温下下保存备用。a. Prepare the purchased synthetic polymer film into a solution of specific concentration for later use, dissolve the purchased DNA sequence (aminated at one end) in Tris-HCl (pH7.4) buffer solution, and store it at low temperature for later use .
b、将金电极先依次用0.3和0.5μm的铝粉进行抛光处理，再依次放入HNO3:H2O(v/v)＝1:1溶液、乙醇溶液、超纯水中，进行超声波清洗，超声清洗的时间分别为3～5min。b. Polish the gold electrode with 0.3 and 0.5 μm aluminum powder in turn, then put it into HNO 3 :H 2 O(v/v)=1:1 solution, ethanol solution, and ultrapure water in turn, and conduct ultrasonic waves The time for cleaning and ultrasonic cleaning is 3-5 minutes respectively.
c、以处理干净的裸的金电极为工作电极，以含有一定浓度模板分子——叶酸的聚合物溶液为电解液，通过电聚合方法，合成聚合物修饰的金电极。c. Using a clean bare gold electrode as a working electrode, using a polymer solution containing a certain concentration of template molecules—folic acid as an electrolyte, and synthesizing a polymer-modified gold electrode by electropolymerization.
d、将上述得到的聚合物修饰金电极放在含有甲醇和醋酸的溶液中浸泡清洗约20～30min，用以将聚合在聚合物中的模板分子——叶酸洗脱掉，从而得到叶酸印迹聚合物膜修饰的金电极。基于叶酸印迹聚合物膜的电化学传感器制备成功。d. Soak and wash the polymer-modified gold electrode obtained above in a solution containing methanol and acetic acid for about 20-30 minutes, so as to elute the template molecule polymerized in the polymer—folic acid, so as to obtain folic acid imprinted polymerization film-modified gold electrodes. An electrochemical sensor based on folic acid imprinted polymer film was successfully prepared.
e、将不同浓度的叶酸受体加入到含有确定浓度的叶酸溶液中培养约40～60min,待叶酸受体与叶酸结合完全，将制备得到的分子印迹膜修饰的金电极置于叶酸和叶酸受体反应后的溶液中，实现对叶酸受体的间接检测。由于溶液中叶酸的浓度是确定的，那么叶酸受体的浓度越大，能够与溶液中的叶酸结合的就越多，使得溶液中剩余的叶酸的量随着加入叶酸受体的量的变化而变化，最终再结合到分子印迹膜修饰的金电极上的模板分子——叶酸的量就会变化，那么分子印迹膜修饰的金电极上对铁氰化钾/亚铁氰化钾探针的电化学响应就会相应变化。因此该传感器可对不同浓度目标叶酸受体进行定量检测。e. Add different concentrations of folic acid receptors into a folic acid solution containing a certain concentration and incubate for about 40-60 minutes. After the folic acid receptors are completely combined with folic acid, place the prepared molecularly imprinted membrane-modified gold electrode on folic acid and folic acid receptors. In the solution after in vivo reaction, the indirect detection of folic acid receptor is realized. Since the concentration of folic acid in the solution is determined, the greater the concentration of folate receptors, the more folic acid in the solution can be combined, so that the amount of folic acid remaining in the solution varies with the amount of folate receptors added. changes, and the amount of folic acid, the template molecule that is finally bound to the molecularly imprinted membrane-modified gold electrode, will change, so the electric potential of the molecularly imprinted membrane-modified gold electrode to the potassium ferricyanide/potassium ferrocyanide probe The chemical response changes accordingly. Therefore, the sensor can quantitatively detect different concentrations of target folate receptors.
f、将不同浓度的汞离子加入到具有确定浓度的叶酸修饰的探针DNA(叶酸-DNA)溶液中培养30～50min，利用DNA序列中的富胸腺嘧啶序列与汞离子的特异性结合作用，使得叶酸-DNA形成双链结构而能被外切酶III切割，释放叶酸分子，将制备得到的分子印迹膜修饰的金电极放在外切酶III切割后的溶液中，实现对汞离子的间接检测。由于溶液中叶酸-DNA的浓度是确定的，那么分子印迹膜修饰的金电极上对铁氰化钾/亚铁氰化钾探针的电化学响应就会随汞离子浓度变化而相应变化。因此该传感器可对不同浓度目标汞离子进行定量检测。f, adding different concentrations of mercury ions into the folic acid-modified probe DNA (folate-DNA) solution with a definite concentration and culturing for 30 to 50 min, utilizing the specific binding of the thymine-rich sequence in the DNA sequence and mercury ions, The folic acid-DNA forms a double-stranded structure and can be cut by exonuclease III to release folic acid molecules, and the prepared molecularly imprinted membrane-modified gold electrode is placed in the solution after exonuclease III cleavage to realize the indirect detection of mercury ions . Since the concentration of folic acid-DNA in the solution is determined, the electrochemical response of the molecularly imprinted membrane-modified gold electrode to the potassium ferricyanide/potassium ferrocyanide probe will change with the concentration of mercury ions. Therefore, the sensor can quantitatively detect target mercury ions with different concentrations.
本发明制备的电化学生物传感器，结合了电化学生物传感器和分子印迹技术两者固有的优势，且合成方法简单、耗能低、成本低。利用目标分析物与模板分子——叶酸的特异性结合作用，或通过外切酶III的切割作用释放模板分子，达到分别检测叶酸受体和汞离子的目的。The electrochemical biosensor prepared by the invention combines the inherent advantages of both the electrochemical biosensor and the molecular imprinting technology, and has a simple synthesis method, low energy consumption and low cost. The purpose of detecting folate receptor and mercury ion is achieved by using the specific binding effect of the target analyte on the template molecule - folic acid, or releasing the template molecule through the cleavage of exonuclease III.
上面结合附图对本发明进行了示例性描述，显然本发明具体实现并不受上述方式的限制，只要采用了本发明的方法构思和技术方案进行的各种改进，或未经改进直接应用于其它场合的，均在本发明的保护范围之内。The present invention has been exemplarily described above in conjunction with the accompanying drawings. Obviously, the specific implementation of the present invention is not limited by the above methods, as long as the various improvements of the method concept and technical solutions of the present invention are adopted, or directly applied to other Occasions, all within the protection scope of the present invention.
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