CN107737944A - A kind of preparation method of golden nanometer particle graphene quantum dot chiral dimer - Google Patents
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Abstract
A kind of preparation method of golden nanometer particle graphene quantum dot chiral dimer, belongs to technical field of material chemistry.The present invention includes the synthesis of golden nanometer particle and graphene quantum dot, golden nanometer particle and the DNA molecular of modified by graphene quantum dot complementation, golden nanometer particle and graphene quantum dot are assembled into dimeric structure, and the transmission electron microscope of dimer characterizes and the measure of chiral signal.The invention provides the preparation method of a kind of golden nanometer particle and graphene quantum dot dimeric structure, and the dimeric structure prepared using DNA Complementary hybridizations is homogeneous, and measures dimer assembly with stable chiral signal by circular dichroism instrument.
Description
Technical field
The invention belongs to technical field of material chemistry, and in particular to a kind of golden nanometer particle-graphene quantum dot chiral dimer
Preparation method.
Background technology
Graphene（Graphene）Be it is a kind of by carbon atom sp2 hybridized orbits form hexangle type be in honeycomb lattice flat thin
Film, the two-dimensional material of only one carbon atom thickness.Because it has unique electrical and optical properties, have in many aspects
Wide application prospect.But graphene is a kind of semiconductor of zero band gap, without band gap, can not produce fluorescence, and
With certain cytotoxicity, so that it is subject to many limitations in the application of field of medicaments.Graphene quantum dot is in stone
A kind of semiconductor-quantum-point for forming size reduction to nanoscale on the basis of black alkene, it can launch fluorescence and cell
Toxicity is smaller, expands its application.
Graphene quantum dot has big specific surface area, low bio-toxicity, good biocompatibility, stronger fluorescence etc.
Characteristic so that it has played great effect in bio-imaging, biology sensor, medicament transport etc..Circular dichroism spectra conduct
A kind of important spectral quality for studying the structure of matter, has carried out in nano science field and has been widely applied, nowadays studied
Going out the assembly of a variety of asymmetric homogeneous nano structures and hetero nano structure has circular dichroism, and graphene quantum dot and
The heterogeneous dissymmetrical structure that golden nanometer particle is assembled into is not studied also.
The content of the invention
It is an object of the invention to provide a kind of golden nanometer particle with chiral acti ve-graphene quantum dot preparation method,
The dimer assembly structure prepared is homogeneous, circular dichroism signal stabilization.
Technical scheme：The invention discloses a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer,
Include the synthesis of 18nm golden nanometer particles, the synthesis of 5nm graphene quantum dots, golden nanometer particle and graphene quantum dot are repaiied respectively
Complementary DNA molecular is adornd, golden nanometer particle and graphene quantum dot form dimeric structure, the transmission electron microscope of dimer
Characterize the measure with chiral signal.
18nm Jenners in a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer of the present invention
The synthesis step of rice corpuscles is：First by 200mL conical flask chloroazotic acid soaked overnight, then cleaned up, added with ultra-pure water
Enter 95.5mL ultra-pure waters in conical flask, add the chlorauric acid solution that 2.5mL mass concentrations are 0.4%, be placed on heating magnetic
It is heated to seething with excitement on power agitator, after constantly boiling 15min, then 2mL mass concentrations is rapidly added in the state of heating stirring
For 1% citric acid three sodium solution, after solution from it is colourless be changed into claret after continuous heating 5min, continue to stir after stopping heating
Room temperature is cooled to solution.
Described in a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer of the present invention
The synthesis step of 5nm graphene quantum dots is：Graphene oxide is heated into 3h for 300 DEG C in the presence of nitrogen protection, so as to
To the graphene film of thermal reduction, weigh 80mg graphene films and be added to containing the dense H of 10mL2SO4With the dense HNO of 30mL3Mixed acid it is molten
Ultrasound overnight in liquid, adds 500mL distilled water diluting, and acid solution is removed with 0.22 μm of filtering with microporous membrane, then with distilling
Water is cleaned 3 times, and finally filter cake is resuspended with 50mL ultra-pure waters, and adjusts pH to 8.0 with NaOH solution, is placed on reaction under high pressure
In kettle, heating response 12h under conditions of 200 DEG C, after being cooled to room temperature, solution is removed with 0.22 μm of filtering with microporous membrane again
The graphene film of bulk is removed, filtrate is collected, will be dialysed 3 times under the conditions of filtrate lucifuge, so as to obtain graphene quantum dot solution.
Gold described in a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer of the present invention
The step of Nanoparticle Modified DNA1 is：The golden nanometer particle newly synthesized is centrifuged into 7min under conditions of 8000r/min, then
Golden nanometer particle is resuspended with pH7.2 0.01M PBSs, and concentrated twice, measures the concentration of golden nanometer particle
For 10nM, then add final concentration of 30nM DNA1, after being incubated 6h at room temperature, by nano-particle 7000r/min bar
10min is centrifuged under part, supernatant is removed, nano-particle is resuspended with pH8.0 0.01M TE buffer solutions, obtains DNA1 modifications
Golden nanometer particle；
DNA1 sequence is：5’-NH2-AAAAAATGGCATGCC-3’。
Stone described in a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer of the present invention
The step of black alkene quantum dot modifying DNA 2 is：5nm graphene quantum dots are centrifuged into 10min under conditions of 12000r/min, so
Graphene quantum dot is resuspended with pH6.5 0.01M MES buffer solutions afterwards, the concentration for measuring graphene quantum dot is 10nM, then
Add n-hydroxysuccinimide and carbodiimide so that graphene quantum dot, n-hydroxysuccinimide, carbodiimide reaction
Mol ratio be 1: 400:1000, at room temperature oscillating reactions 2h make it that the carboxyl of graphene quantum dot is activated, Ran Houli
The heart removes supernatant, and graphene quantum dot is resuspended with pH7.2 0.01M PBSs, adds DNA2 and make it that it is dense eventually
Spend for 30nM, at room temperature after oscillating reactions 4h, 10min is centrifuged under conditions of 12000r/min, is delayed with pH8.0 0.01M TE
Nano-particle is resuspended fliud flushing, obtains the graphene quantum dot of DNA2 modifications；
DNA2 sequence is：5’-NH2-AAAAAGGCATGCCAT-3’。
Gold described in a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer of the present invention
Nano-particle and graphene quantum dot form the step of dimeric structure and are：By the DNA1 of the preparation golden nanometer particle modified and
The graphene quantum dot of DNA2 modifications mixes in equal volume, is incubated 2h at room temperature so that DNA1 and graphite on golden nanometer particle
DNA2 Complementary hybridizations on alkene quantum dot.
Note：DNA of the present invention is synthesized by Shanghai Sheng Gong bioengineering Co., Ltd.
Beneficial effect：Golden nanometer particle and graphene quantum dot are assembled into asymmetry by the present invention using DNA Complementary hybridizations
Dimeric structure, it is homogeneous to dimeric structure by transmission electron microscope observation, and dimerization is measured by circular dichroism instrument
Body assembly has stable chiral signal.
Brief description of the drawings
The TEM figures of Fig. 1 golden nanometer particles-graphene quantum dot dimer.
The circular dichroism figure of Fig. 2 golden nanometer particles-graphene quantum dot dimer.
Embodiment
Embodiment 1
A kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer, includes the synthesis of 18nm golden nanometer particles,
The DNA molecular of the synthesis of 5nm graphene quantum dots, golden nanometer particle and graphene quantum dot difference modification of complementarity, Jenner's grain of rice
Son and graphene quantum dot form dimeric structure, and the transmission electron microscope of dimer characterizes and the measure of chiral signal.
（1）The synthesis of 18nm golden nanometer particles
First by 200mL conical flask chloroazotic acid soaked overnight, then cleaned up with ultra-pure water, add 95.5mL ultra-pure waters in
In conical flask, the chlorauric acid solution that 2.5mL mass concentrations are 0.4% is added, is placed on heating magnetic stirring apparatus and is heated to
Seethe with excitement, after constantly boiling 15min, then the trisodium citrate that 2mL mass concentrations are 1% is rapidly added in the state of heating stirring
Solution, after solution from it is colourless be changed into claret after continuous heating 5min, stop continuing stirring after heating to be cooled to room temperature to solution.
（2）The synthesis of 5nm graphene quantum dots
Graphene oxide is heated into 3h for 300 DEG C in the presence of nitrogen protection, so as to the graphene film heat-treated, weighed
80mg graphene films are added to containing the dense H of 10mL2SO4With the dense HNO of 30mL3Mixed acid solution in overnight ultrasound, add 500mL
Distilled water diluting, remove acid solution with 0.22 μm of filtering with microporous membrane, then cleaned 3 times with distilled water, finally use filter cake
50mL ultra-pure waters are resuspended, and adjust pH to 8.0 with NaOH solution, are placed in autoclave, are heated under conditions of 200 DEG C
12h is reacted, after being cooled to room temperature, solution is removed into the graphene film of bulk with 0.22 μm of filtering with microporous membrane again, collected
Filtrate, it will be dialysed 3 times under the conditions of filtrate lucifuge, so as to obtain graphene quantum dot solution.
（3）Golden nanometer particle modifying DNA 1
The golden nanometer particle newly synthesized is centrifuged into 7min under conditions of 8000r/min, then delayed with pH7.2 0.01M PBS
Golden nanometer particle is resuspended fliud flushing, and is concentrated twice, and the concentration for measuring golden nanometer particle is 10nM, then adds final concentration
For 30nM DNA1, after being incubated 6h at room temperature, nano-particle is centrifuged into 10min under conditions of 7000r/min, in removal
Clearly, nano-particle is resuspended with pH8.0 0.01M TE buffer solutions, obtains the golden nanometer particle of DNA1 modifications；
DNA1 sequence is：5’-NH2-AAAAAATGGCATGCC-3’。
（4）Modified by graphene quantum dot DNA2
5nm graphene quantum dots are centrifuged into 10min under conditions of 12000r/min, then buffered with pH6.5 0.01M MES
Graphene quantum dot is resuspended liquid, and the concentration for measuring graphene quantum dot is 10nM, then add n-hydroxysuccinimide and
Carbodiimide so that graphene quantum dot, n-hydroxysuccinimide, the mol ratio of carbodiimide reaction are 1: 400:1000,
Oscillating reactions 2h make it that the carboxyl of graphene quantum dot is activated at room temperature, is then centrifuged for removing supernatant, with pH7.2's
Graphene quantum dot is resuspended 0.01M PBSs, is added DNA2 and is caused its final concentration of 30nM, vibrates at room temperature
After reacting 4h, 10min is centrifuged under conditions of 12000r/min, nano-particle is subjected to weight with pH8.0 0.01M TE buffer solutions
It is outstanding, obtain the graphene quantum dot of DNA2 modifications；
DNA2 sequence is：5’-NH2-AAAAAGGCATGCCAT-3’。
（5）Golden nanometer particle-graphene quantum dot forms dimeric structure
The graphene quantum dot of the DNA1 of the preparation golden nanometer particles modified and DNA2 modifications is mixed in equal volume, incubated at room temperature
Educate 2h so that the DNA2 Complementary hybridizations on the DNA1 and graphene quantum dot on golden nanometer particle.
（6）The transmission electron microscope of dimer characterizes and the measure of chiral signal
Dimeric structure is characterized using model JEOL JEM-2100 transmission electron microscope, after assembling first
Dimerization liquid solution be added drop-wise to carbon film copper mesh support film on, after being dried under infrared lamp, accelerating potential be 200kV condition
Under characterized.Take 100 μ L dimerization liquid solution to be added in cuvette, use model Bio-Logic MOS-450 circle
Two chromatographs carry out the measure of circular dichroism spectra signal.
Claims (6)
1. the preparation method of a kind of golden nanometer particle-graphene quantum dot chiral dimer, it is characterised in that including 18nm Jenners
The DNA of the synthesis of rice corpuscles, the synthesis of 5nm graphene quantum dots, golden nanometer particle and graphene quantum dot difference modification of complementarity
Molecule, golden nanometer particle and graphene quantum dot form dimeric structure, and the transmission electron microscope of dimer characterizes and chirality
The measure of signal.
2. a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer according to claim 1, it is special
Sign is that the synthesis step of described 18nm golden nanometer particles is：First by 200mL conical flask chloroazotic acid soaked overnight, then
Cleaned up with ultra-pure water, add 95.5mL ultra-pure waters in conical flask, add the gold chloride that 2.5mL mass concentrations are 0.4%
Solution, it is placed on heating magnetic stirring apparatus and is heated to seething with excitement, after constantly boiling 15min, then in the state of heating stirring
Be rapidly added 2mL mass concentrations be 1% citric acid three sodium solution, after solution from it is colourless be changed into claret after continuous heating 5min,
Stop continuing to stir to solution after heating being cooled to room temperature.
3. a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer according to claim 1, it is special
Sign is that the synthesis step of described 5nm graphene quantum dots is：Graphene oxide is added for 300 DEG C in the presence of nitrogen protection
Hot 3h, so as to the graphene film heat-treated, weigh 80mg graphene films and be added to containing the dense H of 10mL2SO4It is dense with 30mL
HNO3Mixed acid solution in ultrasound overnight, add 500mL distilled water diluting, acid removed with 0.22 μm of filtering with microporous membrane
Solution, then cleaned 3 times with distilled water, finally filter cake is resuspended with 50mL ultra-pure waters, and pH to 8.0 is adjusted with NaOH solution, will
It is placed in autoclave, heating response 12h under conditions of 200 DEG C, after being cooled to room temperature, by solution again with 0.22 μm
Filtering with microporous membrane removes the graphene film of bulk, collects filtrate, will be dialysed 3 times under the conditions of filtrate lucifuge, so as to obtain graphite
Alkene quantum dot solution.
4. a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer according to claim 2, it is special
Sign is that the step of described golden nanometer particle modifying DNA 1 is：By the golden nanometer particle newly synthesized 8000r/min condition
Lower centrifugation 7min, then golden nanometer particle is resuspended with pH7.2 0.01M PBSs, and concentrated twice, measure gold
The concentration of nano-particle is 10nM, then adds final concentration of 30nM DNA1, and after being incubated 6h at room temperature, nano-particle is existed
10min is centrifuged under conditions of 7000r/min, removes supernatant, nano-particle is subjected to weight with pH8.0 0.01M TE buffer solutions
It is outstanding, obtain the golden nanometer particle of DNA1 modifications；
DNA1 sequence is：5’-NH2-AAAAAATGGCATGCC-3’。
5. a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer according to claim 3, it is special
Sign is that the step of described modified by graphene quantum dot DNA2 is：By 5nm graphene quantum dots 12000r/min condition
Lower centrifugation 10min, then graphene quantum dot is resuspended with pH6.5 0.01M MES buffer solutions, measures graphene quantum dot
Concentration is 10nM, then adds n-hydroxysuccinimide and carbodiimide so that graphene quantum dot, N- hydroxysuccinimidyls acyl are sub-
Amine, the mol ratio of carbodiimide reaction are 1: 400:1000, at room temperature oscillating reactions 2h cause the carboxyl of graphene quantum dot
Activated, be then centrifuged for removing supernatant, graphene quantum dot is resuspended with pH7.2 0.01M PBSs, add
DNA2 causes its final concentration of 30nM, and at room temperature after oscillating reactions 4h, 10min is centrifuged under conditions of 12000r/min, uses
Nano-particle is resuspended pH8.0 0.01M TE buffer solutions, obtains the graphene quantum dot of DNA2 modifications；
DNA2 sequence is：5’-NH2-AAAAAGGCATGCCAT-3’。
6. a kind of preparation method of golden nanometer particle-graphene quantum dot chiral dimer according to claim 1, it is special
Sign is that the step of described golden nanometer particle and graphene quantum dot form dimeric structure is：The DNA1 of preparation is modified
Golden nanometer particle and the graphene quantum dot of DNA2 modifications mix in equal volume, are incubated 2h at room temperature so that on golden nanometer particle
DNA1 and graphene quantum dot on DNA2 Complementary hybridizations.
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