CN106093159A - A kind of preparation method of biosensor based on polypeptide golden nanometer particle detection metal ion - Google Patents
A kind of preparation method of biosensor based on polypeptide golden nanometer particle detection metal ion Download PDFInfo
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- CN106093159A CN106093159A CN201610394505.XA CN201610394505A CN106093159A CN 106093159 A CN106093159 A CN 106093159A CN 201610394505 A CN201610394505 A CN 201610394505A CN 106093159 A CN106093159 A CN 106093159A
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- nanometer particle
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- interdigital electrode
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Abstract
The invention provides the preparation method of a kind of biosensor based on polypeptide golden nanometer particle detection metal ion, belong to bio-sensing detection technique field.This sensor includes gold interdigital electrode, Graphene or carbon nano-tube channel, is connected to golden nanometer particle and peptide molecule in channel surface.Utilize peptide molecule that metal ion specific recognition causes the conductance change of raceway groove carry out metal ion detection.The present invention utilizes electrostatic interaction, adsorbs golden nanometer particle at graphenic surface, and golden nanometer particle is evenly distributed at graphenic surface, density big, can improve the density of polypeptide, and then improve detection sensitivity.Utilizing the sulfydryl on peptide molecule end group C, peptide molecule can be fixed on golden nanometer particle surface easily, the polypeptide coverage rate on golden nanometer particle surface is high.The peptide molecule specific recognition target metal ions of particular design, so that described sensor is high to the selectivity of target metal ions, is suitable for the detection of target metal ions in Complex water body.
Description
Technical field
The invention discloses a kind of biosensor based on polypeptide-golden nanometer particle detection metal ion and preparation side
Method, is mainly used in highly sensitive, the selective enumeration method of metal ion in water body, belongs to bio-sensing detection technique field.
Background technology
Heavy metal pollution in water body not only breaks up ecosystem balance, also constitutes a threat to human life's health, therefore,
Exploitation water body in (weight) metal ion highly sensitive, method for quick is significant.Traditional metal ion detection side
Method mainly has atomic absorption method, inductively coupled plasma emission spectrography, inductively coupled plasma mass spectrometry, ultraviolet spectrometry
Photometry, electrochemical methods.But these methods need more complicated pretreatment process, apparatus expensive (as inductive etc. from
Daughter emission spectrometer, inductively coupled plasma mass spectrometry), sensitivity relatively low (atomic absorption method, ultraviolet spectrophotometry
Detection limit generally μ g/mL level).In recent years, have benefited from the fast development of nano science, material science, occur in that some are new
Metal ion inspection.Such as, Hg based on golden nanometer particle2+Colorimetric methods
(Angew.Chem.Int.Ed.2007,46,4,093 4096), based on quantum dot or the Hg of organic fluorescence probe2+,Pb2+Fluorescence
Analysis method (Angew.Chem.Int.Ed.2008,47,8,386 8389), based on silver or the Hg of gold nano-material2+Surface enhanced
Raman analysis method (Nanoscale, 2012,4,5902-5909).These methods highly sensitive, selectivity good, but these
Method is vulnerable to testing conditions impact, still suffers from huge challenge for actual sample analysis.
In recent years, electronic sensor based on one-dimensional, two-dimension nano materials obtain extensive concern (Adv.Mater.2007,
19,1439–1451).Its Cleaning Principle is when object and sensor sensing Channel Interaction, causes sensitive pathway conductance
Change carries out the detection of object.For realizing the selective enumeration method of object in complex system, generally object will be had spy
The opposite sex antibody of identification ability, DNA strand modify sensitive pathway, carry out the detection of protein, DNA molecular.Research shows one
The polypeptide a bit with special sequence can be with Selective recognition metal ion.Such as, tripeptides GGH can specific recognition Cu2+
(J.Am.Chem.Soc.1998,120,609-610).The present invention is by golden nanometer particle, many to metal ion specific recognition
Peptide modifies Graphene or the carbon nanotube conducting raceway groove of gold interdigital electrode successively, constructs a kind of novel detection metal ion
Biosensor.This sensor has advantages such as low cost, highly sensitive, specificity good, detection speed is fast, and preparation process
Simply, the detection for Heavy Metals in Waters ion provides a kind of new way.
Summary of the invention
The present invention provides a kind of biosensor based on polypeptide-golden nanometer particle detection metal ion and preparation method,
It is Graphene or the carbon nanotube conducting raceway groove that golden nanometer particle, peptide molecule are modified gold interdigital electrode successively, passes through
Specificity between metal ion and peptide molecule interacts, and carries out in water body that metal ion is highly sensitive, quickly detects.
Technical scheme:
A kind of biosensor based on polypeptide-golden nanometer particle detection metal ion, it is characterised in that this bio-sensing
Device agent structure is gold interdigital electrode, and it is sequentially depositing conducting channel, golden nanometer particle and peptide molecule;
Described golden interdigital electrode is deposited on silicon dioxide, Merlon or other insulant;
Described conducting channel be deposited on by graphene oxide gold interdigital electrode finger areas constitute, channel length 100~
300 μm, width is 1~5 μm;
Described golden nanometer particle adsorbs on conducting channel, and particle diameter is 3~20nm;
Described peptide molecule be GGHC, CALNN, CCCCC or other length amino acid sequence be 4~10, at least one end
For the peptide fragment of C, it is bonded in golden nanometer particle surface.
Described golden nanometer particle is Cu2+, Al3+Or Pb2+。
A kind of preparation method of biosensor based on polypeptide-golden nanometer particle detection metal ion, step is as follows:
(1) gold interdigital electrode cleans up with acetone, water, successively then with the concentrated sulphuric acid that volume ratio is 3:1 and hydrogen peroxide
Mixed solution process, clean up with ionized water, nitrogen dries up;
(2) the golden interdigital electrode after step (1) being processed is placed in 3-aminopropyl triethoxysilane 5~30min, go from
Sub-water cleans, and nitrogen dries up;
(3) graphene oxide solution that 10~30 μ L concentration are 20~100 μ g/mL is dripped the finger in gold interdigital electrode
Region, keeps 60min, takes out gold interdigital electrode water and cleans up, dries;
(4) with gold interdigital electrode as working electrode, Ag/Cl as reference electrode, platinum electrode be to electrode, enter in PBS solution
Row electrochemical reduction;
(5) the golden interdigital electrode of step (4) gained is placed in containing many phenyl ring alkylamine solution keeps 30~120min,
Make containing the absorption of many phenyl ring molecule of alkyl amine on gold interdigital electrode surface;
(6) the golden interdigital electrode of step (5) gained is placed in solution of gold nanoparticles holding 120min, makes Jenner's grain of rice
Son absorption is on gold interdigital electrode surface;The concentration of golden nanometer particle is 0.5~2.0nmol/L, and particle diameter is 3~20nm;
(7) by step (6) gold interdigital electrode be placed in the peptide molecule solution that concentration is 1~100 μm ol/L holding 2~
12h, makes peptide molecule be bonded on golden nanometer particle.
The aminoacid sequence of described peptide molecule be GGHC, CALNN, CCCCC or other sequence length be 4~10, extremely
Few one end is the aminoacid peptide fragment of C.
Described is naphthalene methylamine, naphthalene ethylamine or pyrene methylamine containing many phenyl ring alkylamine.
The invention has the beneficial effects as follows:
(1) utilizing electrostatic interaction, adsorb golden nanometer particle at graphenic surface, golden nanometer particle divides at graphenic surface
Cloth is uniform, density is big, can improve the density of polypeptide, and then improve detection sensitivity.
(2) utilize the sulfydryl on peptide molecule end group C, can easily peptide molecule be fixed on golden nanometer particle table
Face, the polypeptide coverage rate on golden nanometer particle surface is high.
(3) the peptide molecule specific recognition target metal ions of particular design, so that described sensor is to target
The selectivity of metal ion is high, is suitable for the detection of target metal ions in Complex water body.
(4) present invention prepare based on polypeptide-golden nanometer particle detection biosensor for detecting metal ion, select
Property good, highly sensitive, simple to operate, detection speed fast.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of sensor of the invention.
Fig. 2 (a) is the present invention under amplifying 80000 times, the scanning electron microscope of the golden interdigitated electrode-golden nanometer particle of sensor
Figure.
Fig. 2 (b) is the present invention under amplifying 160000 times, the scanning electron microscope of the golden interdigitated electrode-golden nanometer particle of sensor
Figure.
Fig. 3 is that sensor of the invention is to Cu2+Response current change curve.
In figure: 1 conducting channel;2Si/SiO2Substrate;3 interfering ions;4 mark metal ions;5 peptide molecules.
Detailed description of the invention
Below in conjunction with accompanying drawing and technical scheme, further illustrate the detailed description of the invention of the present invention.
Embodiment 1
By magnetic sputtering method at Si/SiO2Surface deposition gold interdigital electrode, cleans up, then with acetone, water successively
With concentrated sulphuric acid: hydrogen peroxide (3:1) mixed solution processes 30min, cleans up with ionized water, and nitrogen dries up.
Gold interdigital electrode is placed in 15min in 3-aminopropyl triethoxysilane, after taking-up, puts into rapidly a large amount of deionization
Cleaning up in water, nitrogen dries up.
Configure 40 μ g/mL through ultrasonic homodisperse graphene oxide solution, take and keep in 20 μ L dropping interdigital electrodes
60min, cleans up with water, dries.With interdigital electrode as working electrode, Ag/Cl as reference electrode, platinum electrode be to electrode,
With-1.2V voltage reduction 10min in the PBS solution of 0.1M.
Configuration 1mg/mL naphthalene methylamine N, dinethylformamide solution, be placed in the naphthalene methylamine naphthalene of 1mg/mL by interdigital electrode
Methylamine solution keeps 30min, cleans up with N, dinethylformamide, dry.
Interdigital electrode is placed in the solution of gold nanoparticles of particle diameter 6nm holding 120min, with deionized water clean up,
Dry;Polypeptide (GGHC) solution that interdigital electrode is placed in 100 μMs again keeps 120min, cleans up with deionized water, dries,
I.e. obtain Cu2+Carry out the biosensor detected.
Embodiment 2
By magnetic sputtering method at Si/SiO2Surface deposition gold interdigital electrode, cleans up, then with acetone, water successively
With concentrated sulphuric acid: hydrogen peroxide (3:1) mixed solution processes 30min, cleans up with ionized water, and nitrogen dries up.
Gold interdigital electrode is placed in 15min in 3-aminopropyl triethoxysilane, after taking-up, puts into rapidly a large amount of deionization
Cleaning up in water, nitrogen dries up.
Configure 40 μ g/mL through ultrasonic homodisperse graphene oxide solution, take and keep in 20 μ L dropping interdigital electrodes
60min, cleans up with water, dries.With interdigital electrode as working electrode, Ag/Cl as reference electrode, platinum electrode be to electrode,
With-1.2V voltage reduction 10min in the PBS solution of 0.1M.
Configuration 1mg/mL naphthalene methylamine N, dinethylformamide solution, be placed in the naphthalene methylamine naphthalene of 1mg/mL by interdigital electrode
Methylamine solution keeps 30min, cleans up with N, dinethylformamide, dry.
Interdigital electrode is placed in the solution of gold nanoparticles that particle diameter is 6nm holding 120min, cleans with deionized water dry
Only, dry;Again interdigital electrode is placed in 100 μMs polypeptide (CALNN) solution keep 120min, with deionized water clean up,
Dry, i.e. obtain Al3+Carry out the biosensor detected.
Embodiment 3
By magnetic sputtering method at Si/SiO2Surface deposition gold interdigital electrode, cleans up, then with acetone, water successively
With concentrated sulphuric acid: hydrogen peroxide (3:1) mixed solution processes 30min, cleans up with ionized water, and nitrogen dries up.
Gold interdigital electrode is placed in 15min in 3-aminopropyl triethoxysilane, after taking-up, puts into rapidly a large amount of deionization
Cleaning up in water, nitrogen dries up.
Configure 40 μ g/mL through ultrasonic homodisperse carbon nano-tube solution, take and keep in 20 μ L dropping interdigital electrodes
60min, cleans up with water, dries.
Configuration 1mg/mL pyrene methylamine N, dinethylformamide solution, be placed in the naphthalene methylamine naphthalene of 1mg/mL by interdigital electrode
Methylamine solution keeps 30min, cleans up with N, dinethylformamide, dry.
Interdigital electrode is placed in the solution of gold nanoparticles that particle diameter is 6nm holding 120min, cleans with deionized water dry
Only, dry;Polypeptide (GGHC) solution that interdigital electrode is placed in 100 μMs again keeps 120min, cleans up with deionized water, dries in the air
Dry, i.e. obtain Cu2+Carry out the biosensor detected.
Claims (5)
1. a biosensor based on polypeptide-golden nanometer particle detection metal ion, it is characterised in that this biosensor
Agent structure is gold interdigital electrode, and it is sequentially depositing conducting channel, golden nanometer particle and peptide molecule;
Described golden interdigital electrode is deposited on silicon dioxide, Merlon or other insulant;
Described conducting channel is deposited on the finger areas of gold interdigital electrode and constitutes by graphene oxide, channel length 100~300 μ
M, width is 1~5 μm;
Described golden nanometer particle adsorbs on conducting channel, and particle diameter is 3~20nm;
Described peptide molecule be GGHC, CALNN, CCCCC or other length amino acid sequence be 4~10, at least one end be C's
Peptide fragment, is bonded in golden nanometer particle surface.
2. according to the biosensor described in claim 1, it is characterised in that described golden nanometer particle is Cu2+、Al3+Or Pb2+。
3. the preparation method of a biosensor based on polypeptide-golden nanometer particle detection metal ion, it is characterised in that step
Rapid as follows:
(1) gold interdigital electrode cleans up with acetone, water successively, then mixed with the concentrated sulphuric acid that volume ratio is 3:1 and hydrogen peroxide
Conjunction solution processes, and cleans up with ionized water, and nitrogen dries up;
(2) the golden interdigital electrode after step (1) being processed is placed in 3-aminopropyl triethoxysilane 5~30min, deionized water
Cleaning, nitrogen dries up;
(3) graphene oxide solution that 10~30 μ L concentration are 20~100 μ g/mL is dripped in the finger district of gold interdigital electrode
Territory, keeps 60min, takes out gold interdigital electrode water and cleans up, dries;
(4) with gold interdigital electrode as working electrode, Ag/Cl as reference electrode, platinum electrode be to electrode, carry out electricity in PBS solution
Electronation;
(5) the golden interdigital electrode of step (4) gained is placed in containing many phenyl ring alkylamine solution keeps 30~120min, makes to contain
There is the absorption of many phenyl ring molecule of alkyl amine on gold interdigital electrode surface;
(6) the golden interdigital electrode of step (5) gained is placed in solution of gold nanoparticles holding 120min, makes golden nanometer particle inhale
It is attached to gold interdigital electrode surface;The concentration of golden nanometer particle is 0.5~2.0nmol/L, and particle diameter is 3~20nm;
(7) step (6) gold interdigital electrode is placed in the peptide molecule solution that concentration is 1~100 μm ol/L holding 2~12h, makes
Peptide molecule is bonded on golden nanometer particle.
Preparation method the most according to claim 1, it is characterised in that the aminoacid sequence of described peptide molecule is
GGHC, CALNN, CCCCC or other sequence length are 4~10, at least one end is the aminoacid peptide fragment of C.
5. according to the preparation method described in claim 3 or 4, it is characterised in that described is naphthalene first containing many phenyl ring alkylamine
Amine, naphthalene ethylamine or pyrene methylamine.
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CN106932453A (en) * | 2017-04-01 | 2017-07-07 | 盐城工学院 | A kind of electrode and preparation method thereof and Sensors & Application |
CN108680633A (en) * | 2018-05-21 | 2018-10-19 | 大连理工大学 | A kind of N-CNF/AuNPs based electrochemical bio-sensing methods for hydroxy radical detection |
CN110186902A (en) * | 2019-06-28 | 2019-08-30 | 武汉中科志康生物科技有限公司 | A kind of surface-enhanced Raman sensor detecting mercury ion |
CN110849941A (en) * | 2019-12-17 | 2020-02-28 | 大连理工大学 | Preparation method of resistance-type humidity sensing device based on loose carbon structure and hydrophilic polymer material composition |
CN114384032A (en) * | 2022-01-17 | 2022-04-22 | 云南大学 | Norovirus detection probe, preparation method thereof, norovirus detection kit and method for detecting norovirus for non-diagnostic purposes |
CN114577878A (en) * | 2022-03-04 | 2022-06-03 | 东北林业大学 | Sex pheromone receptor derived peptide capable of effectively monitoring cotton bollworm and biosensor thereof |
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Cited By (7)
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CN106932453A (en) * | 2017-04-01 | 2017-07-07 | 盐城工学院 | A kind of electrode and preparation method thereof and Sensors & Application |
CN108680633A (en) * | 2018-05-21 | 2018-10-19 | 大连理工大学 | A kind of N-CNF/AuNPs based electrochemical bio-sensing methods for hydroxy radical detection |
CN108680633B (en) * | 2018-05-21 | 2019-06-07 | 大连理工大学 | A kind of N-CNF/AuNPs based electrochemical bio-sensing method for hydroxy radical detection |
CN110186902A (en) * | 2019-06-28 | 2019-08-30 | 武汉中科志康生物科技有限公司 | A kind of surface-enhanced Raman sensor detecting mercury ion |
CN110849941A (en) * | 2019-12-17 | 2020-02-28 | 大连理工大学 | Preparation method of resistance-type humidity sensing device based on loose carbon structure and hydrophilic polymer material composition |
CN114384032A (en) * | 2022-01-17 | 2022-04-22 | 云南大学 | Norovirus detection probe, preparation method thereof, norovirus detection kit and method for detecting norovirus for non-diagnostic purposes |
CN114577878A (en) * | 2022-03-04 | 2022-06-03 | 东北林业大学 | Sex pheromone receptor derived peptide capable of effectively monitoring cotton bollworm and biosensor thereof |
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