CN104535568A - Biological colorimetric sensor and preparation method and application thereof - Google Patents
Biological colorimetric sensor and preparation method and application thereof Download PDFInfo
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- CN104535568A CN104535568A CN201510020927.6A CN201510020927A CN104535568A CN 104535568 A CN104535568 A CN 104535568A CN 201510020927 A CN201510020927 A CN 201510020927A CN 104535568 A CN104535568 A CN 104535568A
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- thrombin
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Abstract
The invention relates to a biological colorimetric sensor and a preparation method and application thereof. The method includes the following steps that (1) the DNA sequence (P3, Thrombin-binding aptamer and P4) is dissolved in a buffer solution, and the P3, Thrombin-binding aptamer and P4 are cultivated before use; (2) applied matter is allocated to be at the corresponding concentration; (3) the processed P3, Thrombin-binding aptamer and P4 are placed in the solution to be cultivated so that the biological colorimetric sensor can be formed, a manganese dioxide nanosheet has mimic enzyme activity, ssDNA has an inhibiting effect on the mimic enzyme activity of the manganese dioxide nanosheet, and the method is applied to detecting various biomolecules (HBV gene, Thrombin and Tolemerase).
Description
Technical field
The present invention relates to the biological colorimetric sensor technical field on two-dimension nano materials manganese dioxide nano-plates/DNA basis, be specifically related to build colorimetric sensor that active based on two-dimension nano materials analogue enztme in DNA build for the inhibiting effect of two-dimension nano materials manganese dioxide nano-plates analogue enztme activity for the detection to various biomolecule.
Background technology
21 century is century of life science, and be an extremely important aspect in life science to the research of DNA, the research of DNA biosensor becomes focus.Current various mode is for detecting biomolecule, and the DNA based sensor that appears as of two-dimension nano materials is filled with powerful vitality, is product of the time by the combination of two-dimension nano materials and DNA based sensor.Two dimension manganese dioxide nano-plates due to its good water-soluble, excellent bio-compatibility, be easy to the performances such as modification and begun one's study.Therefore, apply two-dimentional manganese dioxide nano-plates to construct efficient molecular recognition interface there is potential advantage.
Summary of the invention
The object of the present invention is to provide a kind of biological colorimetric sensor, its preparation method and purposes.Be specifically related to the biological colorimetric sensor on manganese dioxide nano-plates/DNA basis, manganese dioxide nano-plates is utilized to have the inhibiting effect of the active and ssDNA of analogue enztme for manganese dioxide nano-plates analogue enztme activity, be applied to various biomolecule (HBV gene, Thrombin, Tolemerase) detection.Utilize ssDNA can suppress the DNA of the characteristic of manganese dioxide nano-plates analogue enztme activity and dsDNA and other secondary structure can away from and recover manganese dioxide nano-plates simulation enzyme viability and construct biological colorimetric sensor, apply two-dimentional manganese dioxide nano-plates and construct the detection that the biological colorimetric sensor of manganese dioxide nano-plates/DNA that efficient molecular recognition interface builds is applied to various target biological molecules.Concrete technical scheme is as follows:
A preparation method for biological colorimetric sensor, comprises the steps:
(1) by DNA sequence dna (P
3, Thrombin-binding aptamer, P
4) be dissolved in buffer solution, to P before use
3, Thrombin-binding aptamer, P
4cultivate;
(2) corresponding concentration is configured to material used;
(3) by process P
3, Thrombin-binding aptamer and P
4put and cultivate in the solution, form biological colorimetric sensor.
Further, in step (1), buffer solution is the PBS buffer solution of pH7.4.
Further, in step (1), DNA sequence dna is dissolved in after in buffer solution and saves backup down at low temperatures; By P before use
3, Thrombin-binding aptamer, P
4incubated at room temperature a period of time is naturally cooled to after 10 minutes in 80-90 DEG C of heating.
Further, in step (2), material used comprises manganese dioxide nano-plates, thrombin, TMB.
Further, in step (3), by process P
3, Thrombin-binding aptamer and P
4what be placed on that step (2) obtains is dissolved with in the solution of finite concentration manganese dioxide nano-plates.
Further, in step (3), at room temperature 5min is cultivated.
Further, in step (3), the Van der Waals adsorption on the base manganese dioxide nano-plates surface between DNA is utilized to form the biological colorimetric sensor on manganese dioxide nano-plates/DNA basis.
A kind of biological colorimetric sensor, adopts the preparation method of above-mentioned biological colorimetric sensor to obtain.
Further, it is manganese dioxide nano-plates/DNA underlying biological colorimetric sensor.
The purposes of above-mentioned biological colorimetric sensor, further, for the detection to biomolecule (HBVgene, Thrombin, Tolemerase).
Compared with currently available technology, the invention provides the biological colorimetric sensor based on manganese dioxide nano-plates/DNA basis, and be applied to various biomolecule (HBV gene, Thrombin, Tolemerase) detection, the present invention uses manganese dioxide nano-plates to have the inhibiting effect of the active and ssDNA of analogue enztme for manganese dioxide nano-plates analogue enztme activity, prepare the biological colorimetric sensor based on manganese dioxide nano-plates/DNA basis, utilize DNA for the adjustable characteristic of manganese dioxide nano-plates analogue enztme, sensor achieves various biomolecule (HBV gene, Thrombin, Tolemerase) sensitivity, specific detection.
Specifically, the preparation method of this biological colorimetric sensor, use unmarked DNA, simple to operate, cost is very low, avoid any chemical labeling and modify result showing this sensor to various biomolecule (HBVgene, Thrombin, Tolemerase) testing result satisfactory, and have simple to operate, highly sensitive, the feature that detectability is low.
Accompanying drawing explanation
Fig. 1 is the optium concentration suppressing manganese dioxide nano-plates analogue enztme activity
Fig. 2 A is the ultraviolet spectrogram under various material existence condition
Fig. 2 B is based on the corresponding fluorescence spectrum of manganese dioxide nano-plates/DNA sensor markless detection HBV gene correlated results figure, variable concentrations HBVgene
Fig. 2 C is based on manganese dioxide nano-plates/DNA sensor markless detection HBV gene correlated results figure, fluorescence intensity and different HBV gene concentration linear relationship
Fig. 2 D investigates based on the selectivity of manganese dioxide nano-plates/DNA sensor
Fig. 3 is the optium concentration suppressing the active aptamers of manganese dioxide nano-plates analogue enztme
Fig. 4 A is the ultraviolet spectrogram under various material existence condition
Fig. 4 B is based on the corresponding fluorescence spectrum of manganese dioxide nano-plates/DNA sensor markless detection Thrombin correlated results figure, variable concentrations Thrombin
Fig. 4 C is based on manganese dioxide nano-plates/DNA sensor markless detection Thrombin correlated results figure, fluorescence intensity and different Thrombin concentration linear relationship
Fig. 4 D investigates based on the selectivity of manganese dioxide nano-plates/DNA sensor
Fig. 5 is the optium concentration suppressing manganese dioxide nano-plates analogue enztme activity
Fig. 6 A is the ultraviolet spectrogram under various material existence condition
Fig. 6 B is based on the corresponding fluorescence spectrum of manganese dioxide nano-plates/DNA sensor markless detection Tolemerase correlated results figure, variable concentrations Tolemerase
Fig. 6 C is based on manganese dioxide nano-plates/DNA sensor markless detection Tolemerase correlated results figure, fluorescence intensity and different Tolemerase concentration linear relationship
Fig. 6 D investigates based on the selectivity of manganese dioxide nano-plates/DNA sensor
Fig. 7 is the biological colorimetric sensor based on manganese dioxide nano-plates/DNA basis, and is applied to biological HBV gene detection schematic diagram
Fig. 8 is the biological colorimetric sensor based on manganese dioxide nano-plates/DNA basis, and is applied to biological Thrombin detection schematic diagram
Fig. 9 is the biological colorimetric sensor based on manganese dioxide nano-plates/DNA basis, and is applied to biological Tolemerase detection schematic diagram
Embodiment
Describe the present invention with reference to the accompanying drawings below, it is a kind of preferred embodiment in numerous embodiments of the present invention.
The biological colorimetric sensor preparation method on the manganese dioxide nano-plates/DNA basis of this preferred embodiment and the step of application as follows:
(1) DNA sequence dna (P will bought
3, Thrombin-binding aptamer, P
4) be dissolved in PBS (pH 7.4) buffer solution, and save backup down at low temperatures, by P before use
3, Thrombin-binding aptamer, P
4incubated at room temperature a period of time is naturally cooled to after 10 minutes in 80-90 DEG C of heating;
(2) to material used as manganese dioxide nano-plates, thrombin, TMB are configured to corresponding concentration;
(3) by process P
3, Thrombin-binding aptamer and P
4be placed in the solution being dissolved with finite concentration manganese dioxide nano-plates, under 3 room temperatures, cultivate 5min, utilize the Van der Waals adsorption on the base manganese dioxide nano-plates surface between DNA to form the biological colorimetric sensor on manganese dioxide nano-plates/DNA basis.
The concentration of HBV gene is different, the amount forming dsDNA is different, the amount being adsorbed on ssDNA on manganese dioxide nano-plates is different, along with the increase of HBV gene concentration, the amount forming dsDNA is more, and the ssDNA be adsorbed on manganese dioxide nano-plates is fewer, manganese dioxide nano-plates analogue enztme activity is stronger, oxidized more of TMB, the uv absorption of generation is stronger, and colorimetric color is darker.Therefore this sensor quantitatively can detect the HBV gene of variable concentrations.
The concentration of Thrombin is different, the amount of the aptamers that Thrombin identifies is different, the amount being adsorbed on aptamers on manganese dioxide nano-plates is different, along with the increase of Thrombin concentration, the aptamers be adsorbed on manganese dioxide nano-plates is fewer, and manganese dioxide nano-plates analogue enztme activity is stronger, oxidized more of TMB, the uv absorption produced is stronger, and colorimetric color is darker.Therefore this sensor quantitatively can detect the Thrombin of variable concentrations.
The concentration of Tolemerase is different, Yan Sheng the probe P of desorption
4amount different, the amount being adsorbed on ssDNA on manganese dioxide nano-plates is different, along with the increase of Tolemerase concentration, the probe P of desorption
4amount more, be adsorbed on the probe P on manganese dioxide nano-plates
4fewer, manganese dioxide nano-plates analogue enztme activity is stronger, and oxidized more of TMB, the uv absorption of generation is stronger, and colorimetric color is darker.Therefore this sensor quantitatively can detect the Tolemerase of variable concentrations.
Above by reference to the accompanying drawings to invention has been exemplary description; obvious specific implementation of the present invention is not subject to the restrictions described above; as long as have employed the various improvement that method of the present invention is conceived and technical scheme is carried out; or directly apply to other occasion, all within protection scope of the present invention without improving.
Claims (10)
1. a preparation method for biological colorimetric sensor, is characterized in that, comprises the steps:
(1) by DNA sequence dna (P
3, Thrombin-binding aptamer, P
4) be dissolved in buffer solution, to P before use
3, Thrombin-binding aptamer, P
4cultivate;
(2) corresponding concentration is configured to material used;
(3) by process P
3, Thrombin-binding aptamer and P
4put and cultivate in the solution, form biological colorimetric sensor.
2. the preparation method of biological colorimetric sensor as claimed in claim 1, is characterized in that, in step (1), buffer solution is the PBS buffer solution of pH7.4.
3. the preparation method of biological colorimetric sensor as claimed in claim 1 or 2, is characterized in that, in step (1), DNA sequence dna is dissolved in after in buffer solution and saves backup down at low temperatures; By P before use
3, Thrombin-binding aptamer, P
4incubated at room temperature a period of time is naturally cooled to after 10 minutes in 80-90 DEG C of heating.
4. the preparation method of the biological colorimetric sensor according to any one of claim 1-3, is characterized in that, in step (2), material used comprises manganese dioxide nano-plates, thrombin, TMB.
5. the preparation method of the biological colorimetric sensor according to any one of claim 1-4, is characterized in that, in step (3), by process P
3, Thrombin-binding aptamer and P
4what be placed on that step (2) obtains is dissolved with in the solution of finite concentration manganese dioxide nano-plates.
6. the preparation method of the biological colorimetric sensor according to any one of claim 1-5, is characterized in that, in step (3), at room temperature cultivates 5min.
7. the preparation method of the biological colorimetric sensor according to any one of claim 1-6, it is characterized in that, in step (3), the Van der Waals adsorption on the base manganese dioxide nano-plates surface between DNA is utilized to form the biological colorimetric sensor on manganese dioxide nano-plates/DNA basis.
8. a biological colorimetric sensor, is characterized in that, adopts the preparation method of the biological colorimetric sensor as described in claim 1-7 to obtain.
9. biological colorimetric sensor as claimed in claim 8, is characterized in that, it is manganese dioxide nano-plates/DNA underlying biological colorimetric sensor.
10. the purposes of biological colorimetric sensor as claimed in claim 8 or 9, is characterized in that, for the detection to biomolecule (HBVgene, Thrombin, Tolemerase).
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105548167A (en) * | 2015-12-29 | 2016-05-04 | 安徽师范大学 | Manganese dioxide sheet mimic enzyme sensor and preparation method thereof as well as T4PNK detection method |
| CN106434852A (en) * | 2016-09-20 | 2017-02-22 | 江南大学 | Method for realizing intracellular telomerase activity detection based on chiral self-assembled nano sensor |
| CN106483110A (en) * | 2016-09-21 | 2017-03-08 | 安徽师范大学 | A kind of biological sensor, its preparation method and purposes |
| CN114689531A (en) * | 2022-03-01 | 2022-07-01 | 武汉轻工大学 | Preparation method and application of manganese nanoflower-aptamer probe |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103336003A (en) * | 2013-06-27 | 2013-10-02 | 江南大学 | Method for oxidizing TMB with MnO2/CuO composite material as mimetic oxidase |
| CN103926245A (en) * | 2014-04-23 | 2014-07-16 | 安徽师范大学 | Deoxyribonucleic acid (DNA)-modified gold nanoparticle colorimetric sensor and preparation method and use thereof |
-
2015
- 2015-01-15 CN CN201510020927.6A patent/CN104535568A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103336003A (en) * | 2013-06-27 | 2013-10-02 | 江南大学 | Method for oxidizing TMB with MnO2/CuO composite material as mimetic oxidase |
| CN103926245A (en) * | 2014-04-23 | 2014-07-16 | 安徽师范大学 | Deoxyribonucleic acid (DNA)-modified gold nanoparticle colorimetric sensor and preparation method and use thereof |
Non-Patent Citations (12)
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105548167A (en) * | 2015-12-29 | 2016-05-04 | 安徽师范大学 | Manganese dioxide sheet mimic enzyme sensor and preparation method thereof as well as T4PNK detection method |
| CN105548167B (en) * | 2015-12-29 | 2019-05-14 | 安徽师范大学 | Manganese dioxide thin slice simulates the detection method of enzyme sensor and preparation method and T4PNK |
| CN106434852A (en) * | 2016-09-20 | 2017-02-22 | 江南大学 | Method for realizing intracellular telomerase activity detection based on chiral self-assembled nano sensor |
| CN106434852B (en) * | 2016-09-20 | 2019-11-12 | 江南大学 | A method of telomerase activation detection intracellular is realized based on chiral self-assembled nanometer sensor |
| CN106483110A (en) * | 2016-09-21 | 2017-03-08 | 安徽师范大学 | A kind of biological sensor, its preparation method and purposes |
| CN106483110B (en) * | 2016-09-21 | 2020-09-08 | 安徽师范大学 | Fluorescent biosensor, preparation method and application thereof |
| CN114689531A (en) * | 2022-03-01 | 2022-07-01 | 武汉轻工大学 | Preparation method and application of manganese nanoflower-aptamer probe |
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Application publication date: 20150422 |