CN104267088A - An electrochemical biosensor for detecting glutathione and a preparing method thereof - Google Patents
An electrochemical biosensor for detecting glutathione and a preparing method thereof Download PDFInfo
- Publication number
- CN104267088A CN104267088A CN201410163474.8A CN201410163474A CN104267088A CN 104267088 A CN104267088 A CN 104267088A CN 201410163474 A CN201410163474 A CN 201410163474A CN 104267088 A CN104267088 A CN 104267088A
- Authority
- CN
- China
- Prior art keywords
- electrode
- glutathione
- chain
- gold electrode
- solution
- 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.)
- Granted
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to an electrochemical biosensor for detecting glutathione and a preparing method thereof. The biosensor is a three-electrode system sensor, wherein the counter electrode is a platinum electrode, the reference electrode is a saturated calomel electrode, and the working electrode is a gold electrode. The electrochemical biosensor and the method are characterized in that: the gold electrode is modified with double-stranded DNA capable of being adopted as a copper nanocluster synthesis template. According to the electrochemical biosensor and the preparing method, high-affinity bonding of the glutathione and a copper ion is utilized, synthesis of copper nanoclusters on surfaces of the electrodes is inhibited, and indirect detection of the glutathione is achieved by performing electrochemical quantitative characterization on the copper nanoclusters. The linear range of detection of the glutathione is 1-1000 nM, and the detection limit is about 0.42 nM. The electrochemical biosensor and the method have advantages of simple operation, low cost, using convenience, high selectivity, and the like, so that the electrochemical biosensor and the method have large potential application value in the fields of biochemical research, clinical analysis, and other fields.
Description
Technical field
The present invention relates to a kind of novel electrochemical Biosensors and preparation method thereof, particularly a kind of electrochemica biological sensor detecting glutathione and preparation method thereof.
background of invention
Glutathione is the small-molecular-weight non-protein sulfhydryl compound that in cell, content is the abundantest, and it is formed through peptide bond condensation by glutamic acid, halfcystine and glycocoll.Glutathione is the correctives of sulfydryl and sulfide in body, maintain the reducing condition of protein sulfhydryl and enzyme activity, anti-oxidant, maintain in living organism internal oxidition reducing environment balance etc. and play an important role.Some research in recent years shows, the change of body glutathion inside concentration develops relevant with the generation of many diseases such as alzheimer disease, parkinsonism, diabetes, atherosclerotic; And the Sensitive Detection of glutathione can provide much important information for the clinical diagnosis of these diseases and treatment.The technology detecting glutathione now mainly comprises high performance liquid chromatography, ultraviolet-heat amount detection method, capillary electrophoresis, mass spectrometry, spectrofluorimetry etc.These methods respectively have superiority in detection sensitivity, selectivity, detection time and stability, but also there is the deficiency such as complex operation, instrument and equipment costliness simultaneously.Therefore, invent a kind of simple, sensitive glutathione new detecting method and seem very urgent.
Electrochemica biological sensor be a class using electrode as signal converter, the biology sensor measured with current potential or electric current, primarily of molecular recognition and information converting member two parts combination form.Electrochemical system realizes inputing or outputing of electric energy by electrode, thus obtains the electric signal of electrode face finish material.In electrochemical research, conventional three-electrode system, comprises working electrode, auxiliary electrode (also claiming electrode) and contrast electrode.Electric current flows through working electrode and auxiliary electrode, and the current potential measured by working electrode is for contrast electrode.Electrochemical method, as an alanysis detection method, has the advantages such as equipment is cheap, highly sensitive, simple and efficient.
Summary of the invention
An object of the present invention is to provide a kind of electrochemica biological sensor detecting glutathione, and this sensor, by the quantitative test to electrode surface copper nano-cluster, realizes the indirect detection for glutathione.
Two of object of the present invention is the preparation method providing this sensor.
For achieving the above object, the present invention adopts following mechanism: design a DNA single chain P1, its 3' end contains sulfydryl, can pass through the effect self assembly of gold-mercapto covalent bond in gold electrode surfaces; Design in addition one with the DNA single chain P2 of P1 strand base complete complementary, both hybridize formation double-strand, become the template that copper nano-cluster synthesizes at electrode surface.Under reductive agent existent condition, bivalent cupric ion in solution is reduced into monovalence cuprous ion, there is disproportionation reaction and be transformed into bivalent cupric ion and zerovalent copper atom in the latter, and enrichment occurs in the major groove position of the double-stranded DNA that zerovalent copper atom can be fixed at electrode surface and final formation copper nano-cluster.Utilize hydrochloric acid by the copper nano-cluster oxidation dissolution of synthesis, and use electrochemical techniques to detect, the quantitatively characterizing to electrode surface copper nano-cluster can be realized.On the other hand, glutathione can copper ion specifically in binding soln, thus suppresses the reduction process of copper ion, the quantity minimizing of the copper nano-cluster finally causing electrode surface synthesize.The electrochemical signals obtained when utilizing electrochemical techniques to carry out quantitatively characterizing to copper nano-cluster is also corresponding to diminish, by analyze electrochemical signals change number, we just can calculate the concentration of glutathione.
According to above-mentioned mechanism, the present invention adopts following technical scheme:
A kind of electrochemica biological sensor detecting glutathione, for three-electrode system sensor, wherein platinum electrode to electrode, contrast electrode is saturated calomel electrode, working electrode is gold electrode, it is characterized in that described gold electrode being modified with the DNA double chain that can synthesize template as copper nano-cluster, and this double-strand complementary structure.
Above-mentioned DNA double chain is formed by the hybridization of P1, P2 chain, and wherein the sequence of P1 chain is: 5'-TACTCATACGCTCATACGTTCATCACGACTAAAAA-C
6the sequence of-SH-3', P2 chain is: 5'-AGTCGTGATGAACGTATGAGCGTATGAGTA-3'.
The principle of design of above-mentioned sequence is that P1-P2 hybridization chain can keep stable duplex structure at electrode surface in experimentation, and double-chain length is no less than 30 bases (ratio of T-A base-pair is greater than 50%), to ensure the efficiency that copper nano-cluster synthesizes.Once the sequence of DNA is devised, its preparation or chemosynthesis complete transferring to nucleic acid Synesis Company of specialty.
According to a preparation method for the biology sensor of above-mentioned detection glutathione, it is characterized in that the working electrode preparing this sensor, concrete steps are:
A) gold electrode processed is placed in 0.5 M H
2sO
4in, in 0 ~ 1.6 V voltage range, carry out cyclic voltammetry scan, sweep speed and be set to 100 mV/s, until reach stable; Dry up stand-by;
B) step a gained gold electrode being immersed in DNA fixes in buffer solution, left at room temperature is after 15 ~ 18 hours, then to be immersed in 1 mM sulfydryl hexanol aqueous solution lucifuge reaction 0.5 ~ 2.0 hour, with ultrapure water, dry up, namely obtain the gold electrode that first chain is modified; Described DNA fix in buffer solution containing concentration be 1 μM first chain, Tris-HCl, the EDTA of 1 mM of 10 mM, the NaCl of the TCEP of 10 mM and 0.1 M, the pH value of solution is 7.4;
C) be immersed in DNA hybridization buffer liquid by the gold electrode through first chain modification of step b gained, 4 C leave standstill after 1 ~ 2 hour and use ultrapure water, dry up, and namely obtain the gold electrode that complete double-stranded DNA is modified; Described DNA hybridization buffer solution is the phosphate buffer of 10 mM pH 7.4, and wherein containing concentration is second chain of 1 μM and the NaCl of 1 M.
The concrete steps of the disposal route of above-mentioned gold electrode are: drip 20 μ L Piranha solution in pending gold electrode surfaces, namely the volume ratio of Nong Liu Suan ﹕ hydrogen peroxide is 3 ﹕ 1, reacts 2 minutes, and clean with ultrapure water, nitrogen dries up; After gold electrode is polished 2 minutes on 5000 order sand paper, be respectively containing granularity 1 μm, 0.3 μm, 0.05 μm aluminium oxide mortar silk on be polished to minute surface successively, then in ethanol, ultrapure water ultrasonic 2 minutes successively, removing impurity.
A detection method for glutathione, adopts the biology sensor of above-mentioned glutathione, it is characterized in that the concrete steps of the method are:
A. appropriate ascorbic acid powder is taken, 10 mM are diluted to ultrapure water, get 40 μ L to mix with 320 μ L reaction buffers, then in this system, add the mixed solution to be measured that 40 μ L contain 10 μMs of copper ions and variable concentrations glutathione, room temperature reaction 20 minutes; Be the MgCl of the MOPS of 20 mM, the NaCl of 300 mM and 2 mM containing concentration in described reaction buffer
2, the pH value of solution is 7.5.
B. the working electrode gold electrode in biology sensor is put at once the mixed solution of step a gained, react 30 minutes under room temperature.With ultrapure water and after drying up with nitrogen, working electrode is immersed in the copper nano-cluster of lysis electrodes surface synthesis in 200 μ L 0.1 M hydrochloric acid solutions.React after 2 hours, above-mentioned solution is mixed with 4.8 mL 0.5 M NaAc-HAc damping fluids, and carries out Electrochemical Detection in this, as electrolyte solution.Electrochemical Detection concrete steps are: first under-1.2 V voltages, deposit 8 minutes, then use differential pulse voltammetry (DPV) to scan and obtain corresponding electrochemical data; Wherein DPV experiment design parameter is: initial potential 0.1 V, stops current potential 0.35 V, pulse-response amplitude 50 mV, recurrence interval 200 m.
The present invention constructs a kind of electrochemica biological sensor detecting glutathione, utilize glutathione and the strong feature of copper ion affinity, the inhibiting effect of being synthesized copper nano-cluster by glutathione is sensitive with Electrochemical Detection, easily Dominant Facies be combined, by the quantitatively characterizing of copper nano-cluster synthesized electrode surface, quick, sensitive detection is carried out to glutathione, has been with a wide range of applications.
Accompanying drawing explanation
When Fig. 1 is the copper ion that there is variable concentrations in the solution, the DPV quantitatively characterizing result of the copper nano-cluster of electrode surface synthesis.Curve be respectively from a to f 0 M, 1 nM, 10 nM, 100 nM, 1 μM and 10 μMs.
The DPV collection of illustrative plates of Fig. 2 for obtaining in detection 1 μM of glutathione and control experiment.A () control group, not containing glutathione in system; B () experimental group, containing 1 μM of glutathione in system.
Fig. 3 for detect variable concentrations glutathione (be respectively from a to g 0 nM, 1 nM, 5 nM, 10 nM, 100 nM, 1 μM and 10 μMs) time the DPV collection of illustrative plates that obtains.
Fig. 4 is the relation between DPV peak point current and glutathione concentrations, insert figure be glutathione concentrations within the scope of 1-1000 nM, the linear relationship between DPV peak point current and glutathione concentrations logarithm value.
Fig. 5 be detection 1 μM of glutathione and 10 μMs of interference amino acid time the DPV collection of illustrative plates that obtains.
specific implementation method
Embodiment one: the gold electrode preparation that double-stranded DNA is modified
Drip 20 μ L Piranha solution (Nong Liu Suan ﹕ hydrogen peroxide=3 ﹕ 1) reaction 2 minutes in pending gold electrode surfaces, clean with ultrapure water, nitrogen dries up.After being polished 5 minutes on 5000 order sand paper by gold electrode, on the silk containing aluminium oxide (granularity is respectively 1 μm, 0.3 μm, 0.05 μm) mortar, be polished to minute surface successively respectively, then in ethanol, ultrapure water ultrasonic 2 minutes successively, removing impurity.The gold electrode handled well is placed on 0.5 M H
2sO
4middle cyclic voltammetry scan.Arrange scanning voltage scope 0 ~ 1.6 V, sweep velocity 100 mV/s, setting 30 circle is swept to cyclic voltammetry curve and stablizes, and dries up, obtain the naked gold electrode that surface treatment is clean, can be used for the modification of sulfydryl DNA with nitrogen.The DNA that this gold electrode immerses containing 1 μM of P1 chain is fixed buffer solution (10 mM Tris-HCl, 1 mM EDTA, 10 mM TCEP and 0.1 M NaCl, pH 7.4) in left at room temperature 15 ~ 18 hours, namely form the good P1 chain self assembled monolayer without non-specific adsorption.Use ultrapure water gold electrode, and dry up with nitrogen, again gold electrode is immersed the hybridization buffer (phosphate buffer of 10 mM pH 7.4 containing 1 μM of P2 chain, NaCl wherein containing 1 M) in, 4 C leave standstill after 1 ~ 2 hour with ultrapure water, and dry up with nitrogen, namely obtain the gold electrode that complete double-stranded DNA is modified.
Embodiment two: the synthesis of copper nano-cluster and electrochemical quantitative characterize
Get 40 μ L 10 mM ascorbic acid solutions and the 320 μ L reaction buffers (NaCl of 20 mM MOPS, 300 mM and the MgCl of 2 mM
2, pH 7.5) and mixing, then in this system, add the copper ion solution of 40 μ L variable concentrations, leave standstill 20 minutes.The gold electrode that double-stranded DNA is modified is put into above-mentioned mixed solution, reacts 30 minutes under room temperature.After reaction terminates, to dry up with nitrogen with ultrapure water electrode, to remove the unnecessary copper ion being adsorbed on electrode surface.Subsequently, electrode is immersed in the copper nano-cluster of lysis electrodes surface synthesis in 200 μ L 0.1 M hydrochloric acid solutions.After 2 hours, above-mentioned solution is mixed with 4.8 mL 0.5 M NaAc-HAc damping fluids, and carries out Electrochemical Detection in this, as electrolyte solution.
Associated nucleic acid sequences is as follows:
P1 chain-ordering is: 5'-TACTCATACGCTCATACGTTCATCACGACTAAAAA-C
6-SH-3'.
P2 chain-ordering is: 5'-AGTCGTGATGAACGTATGAGCGTATGAGTA-3'.
Electrochemical Detection concrete steps: first deposit 8 minutes under-1.2 V voltages, then DPV scanning obtains corresponding electrochemical data; DPV tests design parameter: initial potential 0.1 V, stops current potential 0.35 V, pulse-response amplitude 50 mV, recurrence interval 200 ms.
As shown in Fig. 1 curve a, when there is not copper ion in reaction solution, do not have obvious electrochemical response peak in the DPV collection of illustrative plates finally obtained, this is because now electrode surface does not form copper nano-cluster.And when there is copper ion in reaction solution, DPV collection of illustrative plates obtains an obvious feature redox peak near 0.22 V, and this peak current absolute value increases with copper ion concentration and increases (curve b to f).Above result shows by series of steps such as dissolving with hydrochloric acid, electro-deposition and DPV scannings, can realize the quantitatively characterizing of the copper nano-cluster to electrode surface synthesis.
Embodiment three: the detection of variable concentrations glutathione
Get 40 μ L 10 mM ascorbic acid solutions and the 320 μ L reaction buffers (NaCl of 20 mM MOPS, 300 mM and the MgCl of 2 mM
2, pH 7.5) and mixing, then in this system, add the mixed solution to be measured that 40 μ L contain 10 μMs of copper ions and variable concentrations glutathione (0 nM, 1 nM, 5 nM, 10 nM, 100 nM, 1 μM and 10 μMs), room temperature reaction 20 minutes.Subsequently, the gold electrode that double-stranded DNA is modified is put into above-mentioned mixed solution, react under room temperature after 30 minutes and carry out Electrochemical Detection, concrete detecting step is identical with embodiment two.
The gold electrode that Fig. 2 curve a shows double-stranded DNA modification acts on a period of time and scan the DPV collection of illustrative plates obtained after necessarily processing in the reaction system only containing ascorbic acid and copper ion.Therefrom can find out, the template that the double chain DNA molecule that electrode surface is fixed can synthesize as copper nano-cluster, and the generation of electrode surface copper nano particles can cause occurring obvious electrochemical response in DPV collection of illustrative plates.And when there is 1 μM of glutathione in system, the DPV response finally obtained obviously weakens (Fig. 2 curve b), this is that the copper ion that can exist in solution due to glutathione is combined, and then inhibits copper nano-cluster in the formation of electrode surface.
As shown in Figure 3, along with the raising of glutathione concentrations, in DPV collection of illustrative plates, electrochemical response reduces gradually, and this illustrates the increase along with glutathione concentrations, increasing copper ion is combined with glutathione, and then the copper nano-cluster causing electrode surface to be formed is fewer and feweri.
DPV peak point current and glutathione concentrations are mapped and obtains Fig. 4.In addition, the insertion figure of Fig. 4 shows, and when glutathione concentrations changes between 1 ~ 1000 nM, logarithm value and the DPV peak point current of glutathione concentrations present good linear relationship, the foundation that quantitatively can detect as glutathione.The minimum detectability of glutathione is about 0.42 nM.
Embodiment four: bioelectrochemical sensor specificity research
In order to verify the specificity of this bioelectrochemical sensor, we with other amino acid (alanine, phenylalanine, glutamic acid etc.) in contrast, according to above-mentioned experimental technique, respectively to concentration be 10 μMs interference amino acid carry out Electrochemical Detection.As shown in Figure 5, when containing 1 μM of glutathione in solution, the DPV peak current absolute value of gained is minimum, be about 0.16 μ A, and it is all larger containing DPV peak current absolute value during other interference amino acid, with close during blank, illustrate that this bioelectrochemical sensor has very high selectivity and specificity for glutathione.
Above result shows, this biology sensor has good selectivity and specificity for glutathione, mentality of designing is simple, with low cost, convenient experimental operation, testing result are sensitive, in Biochemical Research and clinical analysis field, have very large potential using value.Not finding patent is carry out detection GSH about the sensor prepared with double-strand modified gold electrode.
<120> detects electrochemica biological sensor of glutathione and preparation method thereof
<160>?2
<210>?1
<211>?35
<212>?DNA
<213> artificial sequence
<400>?1
5'-TACTC?ATACG?CTCAT?ACGTT?CATCA?CGACT?AAAAA-C
6-SH-3'
<210>?2
<211>?30
<212>?DNA
<213> artificial sequence
<400>?2
5'-?AGTCG?TGATG?AACGT?ATGAG?CGTAT?GAGTA-3'
Claims (5)
1. one kind is detected the electrochemica biological sensor of glutathione, for three-electrode system sensor, wherein platinum electrode to electrode, contrast electrode is saturated calomel electrode, working electrode is gold electrode, it is characterized in that described gold electrode being modified with the DNA double chain that can synthesize template as copper nano-cluster, and this double-strand complementary structure.
2. the electrochemica biological sensor of detection glutathione according to claim 1, it is characterized in that described DNA double chain is hybridized by P1, P2 chain and formed, wherein the sequence of P1 chain is: 5'-TACTCATACGCTCATACGTTCATCACGACTAAAAA-C
6the sequence of-SH-3', P2 chain is: 5'-AGTCGTGATGAACGTATGAGCGTATGAGTA-3'.
3. a preparation method for the biology sensor of detection glutathione according to claim 1, is characterized in that the working electrode preparing this sensor, and concrete steps are:
A) gold electrode processed is placed in 0.5 M H
2sO
4in, in 0 ~ 1.6 V voltage range, carry out cyclic voltammetry scan, sweep speed and be set to 100 mV/s, until reach stable; Dry up stand-by;
B) step a gained gold electrode being immersed in DNA fixes in buffer solution, left at room temperature is after 15 ~ 18 hours, then to be immersed in 1 mM sulfydryl hexanol aqueous solution lucifuge reaction 0.5 ~ 2.0 hour, with ultrapure water, dry up, namely obtain the gold electrode that first chain is modified; Described DNA fix in buffer solution containing concentration be 1 μM first chain, Tris-HCl, the EDTA of 1 mM of 10 mM, the NaCl of the TCEP of 10 mM and 0.1 M, the pH value of solution is 7.4;
C) be immersed in DNA hybridization buffer liquid by the gold electrode through first chain modification of step b gained, 4 C leave standstill after 1 ~ 2 hour and use ultrapure water, dry up, and namely obtain the gold electrode that complete double-stranded DNA is modified; Described DNA hybridization buffer solution is the phosphate buffer of 10 mM pH 7.4, and wherein containing concentration is second chain of 1 μM and the NaCl of 1 M.
4. method according to claim 3, it is characterized in that the concrete steps of the disposal route of described gold electrode are: drip 20 μ L Piranha solution in pending gold electrode surfaces, namely the volume ratio of Nong Liu Suan ﹕ hydrogen peroxide is 3 ﹕ 1, react 2 minutes, clean with ultrapure water, nitrogen dries up; After gold electrode is polished 2 minutes on 5000 order sand paper, be respectively containing granularity 1 μm, 0.3 μm, 0.05 μm aluminium oxide mortar silk on be polished to minute surface successively, then in ethanol, ultrapure water ultrasonic 2 minutes successively, removing impurity.
5. a detection method for glutathione, adopts the biology sensor of glutathione according to claim 1, it is characterized in that the concrete steps of the method are:
A) appropriate ascorbic acid powder is taken, 10 mM are diluted to ultrapure water, get 40 μ L to mix with 320 μ L reaction buffers, then in this system, add the mixed solution to be measured that 40 μ L contain 10 μMs of copper ions and variable concentrations glutathione, room temperature reaction 20 minutes; Be the MgCl of the MOPS of 20 mM, the NaCl of 300 mM and 2 mM containing concentration in described reaction buffer
2, the pH value of solution is 7.5.
B) the working electrode gold electrode in biology sensor is put at once the mixed solution of step a gained, react 30 minutes under room temperature.With ultrapure water and after drying up with nitrogen, working electrode is immersed in the copper nano-cluster of lysis electrodes surface synthesis in 200 μ L 0.1 M hydrochloric acid solutions.React after 2 hours, above-mentioned solution is mixed with 4.8 mL 0.5 M NaAc-HAc damping fluids, and carries out Electrochemical Detection in this, as electrolyte solution.Electrochemical Detection concrete steps are: first under-1.2 V voltages, deposit 8 minutes, then obtain corresponding electrochemical data with differential pulse voltammetry scanning; Wherein DPV experiment design parameter is: initial potential 0.1 V, stops current potential 0.35 V, pulse-response amplitude 50 mV, recurrence interval 200 ms.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410163474.8A CN104267088B (en) | 2014-04-22 | 2014-04-22 | Detect electrochemica biological sensor of glutathione and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410163474.8A CN104267088B (en) | 2014-04-22 | 2014-04-22 | Detect electrochemica biological sensor of glutathione and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104267088A true CN104267088A (en) | 2015-01-07 |
| CN104267088B CN104267088B (en) | 2017-10-24 |
Family
ID=52158629
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410163474.8A Expired - Fee Related CN104267088B (en) | 2014-04-22 | 2014-04-22 | Detect electrochemica biological sensor of glutathione and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104267088B (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105628659A (en) * | 2015-12-21 | 2016-06-01 | 大连理工大学 | Method for detecting lead ions and zinc ions |
| CN106093158A (en) * | 2016-05-07 | 2016-11-09 | 上海大学 | Glutathion detection biosensor, its preparation method and detection method thereof |
| CN106282323A (en) * | 2015-05-29 | 2017-01-04 | 南京理工大学 | High-sensitivity DNA fluorescence analysis method for generating copper nanoparticles based on polythymine as template |
| CN106525947A (en) * | 2016-09-21 | 2017-03-22 | 广西师范学院 | Method for detecting glutathione concentration of solution |
| CN109253993A (en) * | 2018-10-25 | 2019-01-22 | 中国石油大学(华东) | Namo fluorescence probe and its preparation method and application |
| CN109338014A (en) * | 2018-10-19 | 2019-02-15 | 深圳市老年医学研究所 | DNA circulation induces open type DNA fluorescence nano robot construction method |
| CN109374698A (en) * | 2018-11-06 | 2019-02-22 | 杭州电子科技大学 | A method of based on the 2.5D electrochemistry fingerprint identification fleece-flower root or RADIX POLYGONI MULTIFLORI PREPARATA |
| CN112342272A (en) * | 2020-11-06 | 2021-02-09 | 济南大学 | Biosensor for detecting glutathione based on DNA nano machine |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101825597A (en) * | 2010-04-20 | 2010-09-08 | 上海大学 | DNA aptamer modified bioelectrochemical sensor and preparation method thereof |
-
2014
- 2014-04-22 CN CN201410163474.8A patent/CN104267088B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101825597A (en) * | 2010-04-20 | 2010-09-08 | 上海大学 | DNA aptamer modified bioelectrochemical sensor and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| JIE LIU,ET AL.: "A simple and sensitive sensor for rapid detection of sulfide anions using DNA-templated copper nanoparticles as fluorescent probes", 《ANALYST》 * |
| XIN-PING WANG,ET AL.: "A novel fluorescence probe of dsDNA-templated copper nanoclusters for quantitative detection of microRNAs", 《RSC ADV.》 * |
| YA-RU LIU,ET AL.: "Label-free dsDNA-Cu NPs-based fluorescent probe for highly sensitive detection of L-histidine", 《TALANTA》 * |
| YIHUI HU,ET AL.: "Double-strand DNA-templated synthesis of copper nanoclusters as novel fluorescence probe for label-free detection of biothiols", 《ANAL. METHODS》 * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106282323B (en) * | 2015-05-29 | 2019-08-09 | 南京理工大学 | High-sensitivity DNA fluorescence analysis method for generating copper nanoparticles based on polythymine as template |
| CN106282323A (en) * | 2015-05-29 | 2017-01-04 | 南京理工大学 | High-sensitivity DNA fluorescence analysis method for generating copper nanoparticles based on polythymine as template |
| CN105628659A (en) * | 2015-12-21 | 2016-06-01 | 大连理工大学 | Method for detecting lead ions and zinc ions |
| CN106093158A (en) * | 2016-05-07 | 2016-11-09 | 上海大学 | Glutathion detection biosensor, its preparation method and detection method thereof |
| CN106093158B (en) * | 2016-05-07 | 2018-10-23 | 上海大学 | Glutathione detection biosensor, preparation method and its detection method |
| CN106525947A (en) * | 2016-09-21 | 2017-03-22 | 广西师范学院 | Method for detecting glutathione concentration of solution |
| CN106525947B (en) * | 2016-09-21 | 2019-01-11 | 广西师范学院 | The method for detecting solution Glutathione peptide concentration |
| CN109338014A (en) * | 2018-10-19 | 2019-02-15 | 深圳市老年医学研究所 | DNA circulation induces open type DNA fluorescence nano robot construction method |
| CN109338014B (en) * | 2018-10-19 | 2020-06-02 | 深圳市老年医学研究所 | DNA circulation induction open type DNA fluorescence nano robot construction method |
| CN109253993A (en) * | 2018-10-25 | 2019-01-22 | 中国石油大学(华东) | Namo fluorescence probe and its preparation method and application |
| CN109374698A (en) * | 2018-11-06 | 2019-02-22 | 杭州电子科技大学 | A method of based on the 2.5D electrochemistry fingerprint identification fleece-flower root or RADIX POLYGONI MULTIFLORI PREPARATA |
| CN112342272A (en) * | 2020-11-06 | 2021-02-09 | 济南大学 | Biosensor for detecting glutathione based on DNA nano machine |
| CN112342272B (en) * | 2020-11-06 | 2022-09-16 | 济南大学 | Biosensor for detecting glutathione based on DNA nano machine |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104267088B (en) | 2017-10-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104267088A (en) | An electrochemical biosensor for detecting glutathione and a preparing method thereof | |
| Yuan et al. | Simultaneously electrochemical detection of microRNAs based on multifunctional magnetic nanoparticles probe coupling with hybridization chain reaction | |
| CN106525940B (en) | Electrochemical method based on tetra- serobilas of G--ferroheme compound and polymerization chain type iodine detection single-stranded target DNA concentration | |
| CN110618185B (en) | Ratiometric electrochemical detection method of ochratoxin A | |
| CN104651491B (en) | DNA tetrahedral nano-structure signal probe and application thereof | |
| CN105821132B (en) | A method of the specific Single stranded DNA concentration of Electrochemical Detection based on exonuclease and nucleic acid probe | |
| CN103063715B (en) | Method for detecting surviving gene based on graphene-gold composite material electrochemical DNA (Deoxyribose Nucleic Acid) biosensor | |
| CN109797200B (en) | Ratio type telomerase activity quantitative detection method | |
| CN103940890B (en) | Preparation method and application of DNA-AuNPs (gold nanoparticles) nanometer network structure | |
| CN104198551B (en) | Make Platinum Nanoparticles and the glass-carbon electrode of multi-walled carbon nano-tubes modification and the method using this electrode detection estradiol | |
| CN102721728A (en) | Method for simultaneously determining Pb<2+> and Hg<2+> based on electrochemical DNA biosensor | |
| CN104020204A (en) | Electrochemical sensor for detecting lead as well as preparation method and application thereof | |
| CN110487867A (en) | The quantitative detecting method of T4 polynueleotide kinase | |
| CN109706225A (en) | Electrochemical detection method of the palladium nano-particles based on rolling circle amplification mediation to microRNA | |
| CN108398478B (en) | Construction method and application of multifunctional electrochemical sensor based on adenine/Au (III) compound | |
| CN110106232A (en) | Based on target catalysis without the unmarked double tail hybrid organisms sensors of enzyme and preparation method | |
| CN109613095A (en) | Terminal enzyme (DNA) electrochemica biological sensor preparation method and application based on i-motif change of configuration | |
| CN105567808B (en) | The copper nano particles synthetic method and its application in Electrochemical Detection that rolling circle amplification product is template | |
| CN117368284A (en) | Electrochemical sensor for detecting exosome miRNA, and preparation method and application thereof | |
| CN108051492A (en) | A kind of method based on controllable self assembly forceps constructions Electrochemical Detection single-stranded target DNA | |
| CN208860792U (en) | A kind of sensor detecting l-cysteine | |
| CN105044194A (en) | Method used for detecting acrylamide concentration of solutions | |
| CN107228892B (en) | Electrochemistry mercury ion sensor of temperature-controllable and preparation method thereof | |
| CN108426932B (en) | A kind of electrochemica biological sensor and preparation method based on three chain catenated DNAs | |
| CN105259231A (en) | Electrochemical aptamer electrode for terramycin detection and preparation method thereof |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20171024 Termination date: 20200422 |