CN104165855A - Specific polypeptide modified colorimetric sensor and making method thereof - Google Patents
Specific polypeptide modified colorimetric sensor and making method thereof Download PDFInfo
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- CN104165855A CN104165855A CN201410193866.9A CN201410193866A CN104165855A CN 104165855 A CN104165855 A CN 104165855A CN 201410193866 A CN201410193866 A CN 201410193866A CN 104165855 A CN104165855 A CN 104165855A
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Abstract
The invention relates to a specific peptide modified colorimetric sensor and a making method thereof. The colorimetric sensor is used for detecting copper ions. The sensor uses a gold-mercapto key to make the surface of gold nanoparticles to be modified by a specific peptide, the addition of copper ions to be measured changes the conformation of the specific polypeptide from alpha-helix to beta-folding, and the gold nanoparticles mutually aggregate by using beta-folding fragments formed by the surface modification specific polypeptide in order to change the absorbance of a solution and generate color difference. The copper ions are detected by the gold nanoparticles modified by the specific peptide for the first time, and the colorimetric sensor of the gold nanoparticles modified by the specific polypeptide has very high sensitivity, simplicity and specificity on the copper ions, has a wide detection range, has potential application values in the detection of the content of the copper ions in drinking water, serum, rivers, sewage and other solutions, and also has the advantages of convenient operation, high sensitivity and wide application prospect.
Description
Technical field
The present invention relates to a kind of colorimetric sensor and preparation method thereof, colorimetric sensor that particularly a kind of specific polypeptide is modified and preparation method thereof.
Background technology
Copper is essential a kind of trace heavy metal element in animal and plant body.In body, appropriate copper has important effect in the physiology courses such as energetic supersession, oxygen transportation and signal transduction.But copper will bring harm to life body intensive amount is too high, in body, too much copper may cause a series of nerve degenerative diseases, such as: remote gram this syndrome, Wilson Cotard and Alzheimer's disease etc.The USEPA safety margin of regulation copper in drinking water constituent content is 1.33ppm(approximately 20 μ M), China National Standards Commission and the Ministry of Public Health have also stipulated that copper in drinking water ion concentration is not higher than 1.0mg/L(approximately 15.625 μ M in the < < drinking water sanitary standard > > that combines issue for 2007).As can be seen here, how the copper ion in detection potable water, sewage, river and biosome rapidly and efficiently just seems very important and urgent.
Colorimetric sensor is that a class is measured and supervene the sensor of color distortion with absorbance change or crest displacement.Colorimetric sensor take predetermined substance in certain wavelength coverage to electromagnetic absorption characteristic be that it is easy and simple to handle because having, with low cost, highly sensitive according to the qualitative and quantitative testing tool of setting up, the advantage such as high specificity and fast detecting is used widely.
In the last few years, nano material technology developed and was widely used in the fields such as detection of heavy metal ion, drug delivery, medical diagnosis on disease rapidly.Nano material itself has the good characteristics such as surface effect, micro-size effect, quantum effect and macro quanta tunnel effect, makes it in Development of Novel high sensitivity, high stability, low-cost bio sensor field, become domestic and international study hotspot.Especially gold nano-material, because it has excellent optical property in visible-range, and becomes the focus material of research, the value that has a wide range of applications, and the existing business-like sensor based on gold nano-material is in market sale, as early pregnancy test paper at present.Because of it, can there is specific effect by material (as ion, little molecule, protein, nucleotide etc.) corresponding thereto in specific polypeptide, can covalent bond form (as gold-mercapto key and amido link) modify to other materials again simultaneously, and be subject to researcher's favor, make it in field of biosensors, there is great application prospect, at present researcher mainly utilizes the colorimetric detection system that its characteristic goes design construction to detect for enzyme biopsy survey and specific protein, but heavy metal ion particularly the context of detection of copper ion have not been reported.
Summary of the invention
The colorimetric sensor that provides a kind of specific polypeptide to modify is provided one of object of the present invention.In order to realize the detection to copper ion.
Two of object of the present invention is to provide the preparation method of this sensor.
For achieving the above object, the present invention adopts following mechanism: first by gold-mercapto key, specific polypeptide is modified to gold nano grain surface, under the condition existing at copper ion, impel the conformation of the specific polypeptide on gold nano grain surface to change β-pleated sheet into by α spiral, thereby cause the reduction of gold nano grain dispersiveness and directly cause between gold nano grain and assemble, showing as in the reduction of 520nm wavelength place system absorbance and the change of color.
The colorimetric sensor that specific polypeptide is modified, is characterized in that this colorimetric sensor is on gold nano grain surface, by gold-mercapto key, is modified with specific polypeptide, and described specific polypeptide and the mass ratio of gold nano grain are: 1:180-1:200; The particle diameter of described gold nano grain: 12.5-13nm; The sequence of described specific polypeptide is: Ac – Cys-Gly-Gly-Gly-Ser-Ile-Arg-Lys-Leu-Glu-Tyr-Glu-Ile-Glu-Glu-Leu-Arg-Leu-Arg-Ile-Gly.
A method of preparing the colorimetric sensor that above-mentioned specific polypeptide modifies, is characterized in that the concrete steps of the method are: gold nano grain is dispersed in and in deionized water, is mixed with the suspending liquid that concentration is 2.3-3.0nM/L; By the mass ratio of described specific polypeptide and gold nano grain, be: the ratio of 1:180-1:200, the solution of the specific polypeptide that is 0.02mg/mL by concentration and described gold nano grain suspending liquid mix rear incubated at room and spend the night; After the centrifugal 20min of centrifugal separator, remove supernatant, obtain the gold nano grain that specific polypeptide is modified.
The preparation method of above-mentioned gold nano grain is: after the chlorauric acid solution that is 0.01% by mass concentration is heated with stirring to and refluxes 2～3 minutes, adding mass concentration is 1% citric acid three sodium solution, after reactant liquor color gradually becomes claret, stop heating and continue to be stirred to room temperature; Described gold chloride and the mass ratio of trisodium citrate are 1:0.3-1:0.5.
Finally, by potpourri centrifugal (10000rpm 20min) again, with HEPES solution, redissolve, obtain the gold nano grain of 1mL modified specificity polypeptide.With ultraviolet spectrophotometer, detect respectively the UV spectrum of the gold nano grain of naked gold nano grain and modified specificity polypeptide.
UV spectrum under the copper ion of variable concentrations exists: the gold nano grain of getting the modified specificity polypeptide of a plurality of 45 μ L, the copper ion solution that adds respectively wherein 5 μ L variable concentrations, mix, hatch after 90min with liquid-transfering gun and join and in cuvette, measure UV spectrum.Can see the increase along with copper ion concentration, in 520nm place absorbance, decline also larger, and peak position side-play amount increases, and copper ion concentration during lower than 1 μ M absorbance change relatively littlely, when copper ion concentration is greater than 150 μ M, matched curve tends towards stability.Through being that the absorbance variable quantity of the gold nano grain modified of specific polypeptide in 10-150 μ M at 520nm place carries out linear fit to copper ion concentration scope, the accurate range of linearity that obtains detecting copper ion is 10-150 μ M, coefficient R
2be 0.99853, illustrate that the gold nano grain that specific polypeptide is modified can detect the copper ion of 10-150 μ M concentration range.
Advantage and disadvantage of the present invention is as described below:
The gold nano grain that the present invention modifies specific polypeptide is for the first time applied to the detection of copper ion, observe the variation of gold nano grain optical characteristics before and after modifying, the gathering situation that copper ion causes the gold nano grain that specific polypeptide is modified is passed in research in time, cause that with the copper ion of variable concentrations the absorbance of the gold nano grain that specific polypeptide is modified changes and color distortion, observe the gold nano grain of unmodified specific polypeptide and the gold nano grain of the modified specificity polypeptide aggreation under copper ion exists simultaneously and cause the difference of absorbance and color distortion, and the high selectivity of the gold nano grain colorimetric sensor of specific polypeptide modification to copper ion detection.Result shows, the gold nano grain that specific polypeptide is modified can detect copper ion effectively, at copper ion concentration, is to have good linear relationship, coefficient R within the scope of 10-150 μ M
2be 0.99853.
The present invention has built a kind of novel sensor detecting for copper ion, and the gold nano grain that specific polypeptide is modified is applied to the colorimetric detection of copper ion, has obtained satisfied result.Same principle also extends to the detection of other material, and application prospect is extensive.
The gold nano grain colorimetric sensor that specific polypeptide prepared by the inventive method is modified is applied to the detection of copper ion, the advantage such as have that convenient experimental operation, sensitivity are high, high specificity, sensing range are wide.Can successful detectable concentration scope the copper ion that is 10-150 μ M, the least concentration that can detect is 1 μ M.The colorimetric sensor that the gold nano grain that the present invention modifies specific polypeptide first detects as copper ion, detection to content of copper ion in the liquid such as potable water, serum, river, sewage has potential using value, same principle can be for the detection of other ions, and application prospect is extensive.
Accompanying drawing explanation
Fig. 1 is that the copper ion of variable concentrations and the gold nano grain of specific polypeptide modification are hatched the UV spectrogram after 90min.
Embodiment
After now specific embodiments of the invention being described in.
embodiment 1:
The gold nano grain that in the present embodiment, specific polypeptide is modified and UV spectrometric method and the step of copper ion effect are as follows:
(1) preparation of the gold nano grain of specificity modified polypeptide: first, prepare the gold nano grain that particle diameter is 13nm.The chlorauric acid solution that is 0.01% by mass concentration joins in the round-bottomed flask that reflux condensate device is housed, heating rapid stirring, when chlorauric acid solution boiling is after 2-3 minute, disposablely add fast the citric acid three sodium solution that 3.5 mL mass concentrations are 1%, continuous heating is 15 minutes again, after solution colour gradually becomes claret, stops heating and continue stirring 30min, make it be cooled to room temperature, in 4 ℃, store for future use.Then, get specific polypeptide solution that 20 μ L concentration are 1mg/mL and join in 980 μ L HEPES solution and mix, obtain the modification solution of 1mL gold nano grain.Get above-mentioned gold nano grain solution 1mL, with supercentrifuge, carry out centrifugal (12000rpm 20min), remove supernatant, then add isopyknic peptide modified liquid to redissolve, overnight incubation under room temperature.Finally, by potpourri centrifugal (12000rpm 20min) again, with HEPES solution, redissolve, obtain the gold nano grain of 1mL modified specificity polypeptide.With ultraviolet spectrophotometer, detect respectively the UV spectrum of the gold nano grain of naked gold nano grain and modified specificity polypeptide.Can find, there is skew in the peak position of gold nano grain before and after specific polypeptide is modified, and the particle diameter of the gold nano grain of specific polypeptide modification is obviously greater than naked gold nano grain.
(2) the UV spectrum under the existence of the copper ion of variable concentrations: the gold nano grain of getting the modified specificity polypeptide of a plurality of 45 μ L, the copper ion solution that adds respectively wherein 5 μ L variable concentrations, mix, hatch after 90min with liquid-transfering gun and join and in cuvette, measure UV spectrum.Can see the increase along with copper ion concentration, in 520nm place absorbance, decline also larger, and peak position side-play amount increases, and copper ion concentration during lower than 1 μ M absorbance change relatively littlely, when copper ion concentration is greater than 150 μ M, matched curve tends towards stability.Through being that the absorbance variable quantity of the gold nano grain modified of specific polypeptide in 10-150 μ M at 520nm place carries out linear fit to copper ion concentration scope, the accurate range of linearity that obtains detecting copper ion is 10-150 μ M, coefficient R
2be 0.99853, illustrate that the gold nano grain that specific polypeptide is modified can detect the copper ion of 10-150 μ M concentration range.
Colorimetric method for determining copper ion:
Employing copper ion concentration is respectively: 0 μ M, 1 μ M, 10 μ M, 20 μ M, 30 μ M, 50 μ M, 80 μ M, 100 μ M, 150 μ M, 200 μ M.
Test condition: the light that the wavelength of take is 400-800nm is light source, usings 90min as incubation time, carries out the UV spectroscopic assay under the effect of variable concentrations copper ion at ambient temperature.
Referring to accompanying drawing, the gold nano grain that the copper ion that Fig. 1 is variable concentrations and specific polypeptide are modified is hatched the UV spectrogram after 90min.
As seen from the figure: along with the increase (with the direction of arrow) of copper ion concentration, in 520nm place absorbance, decline also larger, and peak position side-play amount increases.
Claims (3)
1. the colorimetric sensor that specific polypeptide is modified, is characterized in that this colorimetric sensor is on gold nano grain surface, by gold-mercapto key, is modified with specific polypeptide, and described specific polypeptide and the mass ratio of gold nano grain are: 1:180-1:200; The particle diameter of described gold nano grain: 12.5-13nm; The sequence of described specific polypeptide is: Ac – Cys-Gly-Gly-Gly-Ser-Ile-Arg-Lys-Leu-Glu-Tyr-Glu-Ile-Glu-Glu-Leu-Arg-Leu-Arg-Ile-Gly.
2. a method of preparing the colorimetric sensor that specific polypeptide according to claim 1 modifies, is characterized in that the concrete steps of the method are: gold nano grain is dispersed in and in deionized water, is mixed with the suspending liquid that concentration is 2.3-3.0nM/L; By the mass ratio of described specific polypeptide and gold nano grain, be: the ratio of 1:180-1:200, the solution of the specific polypeptide that is 0.02mg/mL by concentration and described gold nano grain suspending liquid mix rear incubated at room and spend the night; After the centrifugal 20min of centrifugal separator, remove supernatant, obtain the gold nano grain that specific polypeptide is modified.
3. method according to claim 2, the preparation method who it is characterized in that described gold nano grain is: after the chlorauric acid solution that is 0.01% by mass concentration is heated with stirring to and refluxes 2～3 minutes, adding mass concentration is 1% citric acid three sodium solution, after reactant liquor color gradually becomes claret, stop heating and continue to be stirred to room temperature; Described gold chloride and the mass ratio of trisodium citrate are 1:0.3-1:0.5.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105403558A (en) * | 2015-11-18 | 2016-03-16 | 沈阳药科大学 | Method for on-line and quick detection of lead ions |
| CN105784616A (en) * | 2016-04-08 | 2016-07-20 | 江南大学 | Method for detecting cysteine or acetylcysteine on basis of bimetal nanometer clusters |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100290992A1 (en) * | 2006-08-11 | 2010-11-18 | Frank Seela | Nanoparticle nucleic acid binding compound conjugates forming i-motifs |
| KR20110031689A (en) * | 2009-09-21 | 2011-03-29 | 숭실대학교산학협력단 | Colorimetric sensor possible to detect amino acids using surface-modified gold nanoparticles and detection method of the amino acids and peptide using color recovery of the gold nanoparticles |
| CN102466639A (en) * | 2010-11-12 | 2012-05-23 | 中国科学院大连化学物理研究所 | Method for detecting various heavy metal ions with photochemical colorimetric sensor array |
| CN101710076B (en) * | 2009-12-29 | 2012-07-04 | 东北师范大学 | Lead ion colorimetric detection probes and application method thereof |
| CN102608180A (en) * | 2012-03-21 | 2012-07-25 | 上海大学 | Bioelectrochemical sensor for detecting silver ion and preparation method thereof |
-
2014
- 2014-05-09 CN CN201410193866.9A patent/CN104165855A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100290992A1 (en) * | 2006-08-11 | 2010-11-18 | Frank Seela | Nanoparticle nucleic acid binding compound conjugates forming i-motifs |
| KR20110031689A (en) * | 2009-09-21 | 2011-03-29 | 숭실대학교산학협력단 | Colorimetric sensor possible to detect amino acids using surface-modified gold nanoparticles and detection method of the amino acids and peptide using color recovery of the gold nanoparticles |
| CN101710076B (en) * | 2009-12-29 | 2012-07-04 | 东北师范大学 | Lead ion colorimetric detection probes and application method thereof |
| CN102466639A (en) * | 2010-11-12 | 2012-05-23 | 中国科学院大连化学物理研究所 | Method for detecting various heavy metal ions with photochemical colorimetric sensor array |
| CN102608180A (en) * | 2012-03-21 | 2012-07-25 | 上海大学 | Bioelectrochemical sensor for detecting silver ion and preparation method thereof |
Non-Patent Citations (4)
| Title |
|---|
| TIANRONG LI 等: ""A novel fluorescein derivative as a colorimetric chemosensor for detecting copper(II) ion"", 《DYES AND PIGMENTS》 * |
| XIAORONG HE 等: ""Gold Nanoparticle-Based Fluorometric and Colorimetric Sensing of Copper(II) Ions"", 《ADVANCED MATERIALS》 * |
| 李向华 等: ""银纳米粒子比色传感器的研究进展"", 《广州化工》 * |
| 林奇 等: ""汞离子荧光、比色传感器"", 《化学进展》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105403558A (en) * | 2015-11-18 | 2016-03-16 | 沈阳药科大学 | Method for on-line and quick detection of lead ions |
| CN105403558B (en) * | 2015-11-18 | 2019-04-02 | 沈阳药科大学 | A kind of method of quick detection lead ion |
| CN105784616A (en) * | 2016-04-08 | 2016-07-20 | 江南大学 | Method for detecting cysteine or acetylcysteine on basis of bimetal nanometer clusters |
| CN105784616B (en) * | 2016-04-08 | 2018-07-17 | 江南大学 | The method for detecting cysteine or acetylcysteine based on bimetal nano cluster |
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Application publication date: 20141126 |