CN105327354B - A kind of preparation method of surface-functionalized tobacco mosaic virus (TMV) nanowire composite - Google Patents
A kind of preparation method of surface-functionalized tobacco mosaic virus (TMV) nanowire composite Download PDFInfo
- Publication number
- CN105327354B CN105327354B CN201510696672.5A CN201510696672A CN105327354B CN 105327354 B CN105327354 B CN 105327354B CN 201510696672 A CN201510696672 A CN 201510696672A CN 105327354 B CN105327354 B CN 105327354B
- Authority
- CN
- China
- Prior art keywords
- tmv
- mosaic virus
- tobacco mosaic
- dopamine
- 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.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/02—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0002—General or multifunctional contrast agents, e.g. chelated agents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
Abstract
The invention discloses a kind of preparation method of surface-functionalized tobacco mosaic virus (TMV) nanowire composite.The preparation method comprises the following steps：By while tobacco mosaic virus (TMV) assembles end to end in acid condition, dopamine and oxidant form the tobacco mosaic virus (TMV) nano wire of poly-dopamine cladding in the outer surface in-situ oxidizing-polymerizing of virus protein, then the reproducibility and reactivity of surface poly-dopamine itself are utilized in the basic conditions, without extra reducing agent and chemical modification, the further load gold nano grain of can and functional modification contain the molecule with bioactivity of amino or sulfydryl, obtain surface-functionalized tobacco mosaic virus (TMV) nanowire composite.The present invention has the characteristics of simple technique, convenience and high-efficiency, can be widely used in the biological medical nano field of new materials such as living things catalysis, bio-imaging, cancer target probe and nano-medicament carrier.
Description
Technical field
The present invention relates to nanometer field of new materials.Received more particularly, to a kind of surface-functionalized tobacco mosaic virus (TMV)
The preparation method of rice noodles composite.
Background technology
Nano composite material is a kind of new nano material, both the small-size effect with nano material, high-ratio surface
The advantages that long-pending, diversified shape characteristic and assembling form, again can be with other compound inorganic, organic, macromolecules and biological work(
The property of energy material, is catalyzed in industrial high-efficient, and the new function material field such as bio-medical engineering has obtained extensive hair
Exhibition.Therefore new one-dimensional nano line composite is developed in living things catalysis, bio-imaging, cancer target probe and Nano medication
There is very huge potential using value in terms of carrier.
Poly-dopamine is that oxidation polymerization is formed under alkaline environment by dopamine, is a kind of new organic polymer
Adhesion material, substrate surface can be uniformly coated on, and further can contain amino by functional modification in material surface
Or the molecule of sulfydryl.And existing technology is mainly in inorganic metal, metal oxide, ceramics, hydrophobic macromolecule material
Material and carbon material surface carry out the cladding and functional modification of poly-dopamine.Such as carried in United States Patent (USP) US 2008/0149566
What is gone out coats in a variety of inorganic and poly-dopamine of polymer surface；Chinese patent CN 102813963 and CN
102000658 be in the inorganic of bio-medical, metal, metal alloy and polymer surface poly-dopamine functional modification
Molecule with bioactivity；Chinese patent CN 104258909 is to be born in magnetic Fe_3O_4 nano grain surface with poly-dopamine
Carry gold nano grain；Chinese patent CN 104666251 is in biodegradable biopolymer nanoparticles surface poly-dopamine
Load hydrophobic drug；Chinese patent CN 103920152 and CN 104549159 is in multi-walled carbon nanotube and magnetic silver nanoparticle
Line is the one-dimensional inorganic nano material Surface coating poly-dopamine of representative.But these technical methods are all not involved with hydrophily
Protein surface or by protein assembly virus type biological nano particle surface carry out poly-dopamine cladding and further
Functional modification.
Tobacco mosaic virus (TMV) is a kind of infection tobacco and the plant virus of other Solanaceae classes, by 2130 virus capsid proteins
Assembling forms 300nm length, diameter 18nm nano tubular structure, has the single dispersing characteristic of homogeneous grain diameter, is a kind of preferable
Construct the biological template of monodimension nanometer material.Using tobacco mosaic virus (TMV) as template, the side that is assembled end to end by virus nanoparticles
Formula forms one-dimensional orderly nano thread structure, and in polyaniline, polypyrrole and the polystyrolsulfon acid of outer Surface coating electric conductivity
Sodium high polymer material, prepare the one-dimensional nano line composite with conducting function had document report (such as Z.Niu,
M.Bruckman,et al.Study and characterization of tobacco mosaic virus head-to-
tail assembly assisted by aniline polymerization[J].Chem.Commun.,2006,28:
3019；Z.Niu,et al.Assembly of tobacco mosaic virus into fibrous and
macroscopic bundled arrays mediated by surface aniline polymerization[J]
.Langmuir,2007,23:6719；Z.Niu,et al.Biological templated synthesis of water-
soluble conductive polymeric nanowires[J].Nano letters,2007,7:3729).It is but existing
Covering material used in method is all the macromolecule of electric conductivity, without biocompatibility, can not be obtained in biomedical sector
To application.
In addition, using tobacco mosaic virus (TMV) as template and carrier, in its area load gold nano grain and functional modification point
Although the method for son has been carried out that (such as Chinese patent CN 103706800 is to express to obtain six by the method for molecular biology
The coat protein for mosaic virus of tobacco of histidine modification, the gold nano grain of reducing agent realization is added in tobacco mosaic virus (TMV) table
The load in face；Chinese patent CN 102578695 is attracted each other by electrostatic, with chemical deposition in the presence of reducing agent
Method is in tobacco mosaic virus (TMV) area load gold nano grain；Chinese patent CN 103613647 is by molecular biology
Method is expressed to obtain the coat protein for mosaic virus of tobacco of cysteine mutation, then arrives fluorescence molecule and cancer therapy drug modification
The protein surface of cysteine mutation is prepared for nano-medicament carrier), but all there is operating procedure complexity in these methods, prepare week
Phase is long, it is necessary to which the method modification gene order and expressing protein of molecular biology reduce to obtain gold, it is necessary to it is possible to additionally incorporate reducing agent
The problems such as nano particle and defect.
So far, coated using the poly-dopamine with biocompatibility, using tobacco mosaic virus (TMV) as template group
The preparation method for filling the one-dimensional nano line composite formed was not yet reported that；Simultaneously in this tobacco mosaic virus (TMV) nano wire
Surface, using the reproducibility and reactivity of poly-dopamine itself, without extra reducing agent and molecular biology method, it is possible to
The method that further facilitating efficiently load gold nano grain and functional modification has bioactive molecule also has no report.
The content of the invention
It is an object of the invention to provide a kind of preparation of surface-functionalized tobacco mosaic virus (TMV) nanowire composite
Method.The preparation method operating procedure is simple, short preparation period, is coated using the poly-dopamine with biocompatibility,
The one-dimensional nano line composite formed using tobacco mosaic virus (TMV) as template assemblies；Simultaneously in this tobacco mosaic virus (TMV) nano wire
Surface, using the reproducibility and reactivity of poly-dopamine itself, without extra reducing agent and molecular biology method, it is possible to
Further facilitating efficiently load gold nano grain and functional modification has bioactive molecule.
To reach above-mentioned purpose, the present invention uses following technical proposals：
A kind of preparation method of surface-functionalized tobacco mosaic virus (TMV) nanowire composite, including following prepare walk
Suddenly：The assembling of the tobacco mosaic virus (TMV) nano wire of Surface coating poly-dopamine is first carried out, recycles and is wrapped in gained nanowire surface
The poly-dopamine covered carries out functional modification, obtains surface-functionalized tobacco mosaic virus (TMV) nanowire composite.
Preferably, the assembling of the tobacco mosaic virus (TMV) nano wire of the Surface coating poly-dopamine refers to molten in acidic buffer
Dopamine hydrochloride and oxidant are added in liquid, it is poly- more in tobacco mosaic virus (TMV) Surface Creation by the method for in-situ oxidizing-polymerizing
While bar amine clad, by tobacco mosaic virus (TMV) be template by assembling to form one-dimensional nano wire in a manner of from beginning to end,
And the 1-dimention nano cable architecture to be formed is fixed and maintained to poly-dopamine clad, specifically comprises the following steps：
1) tobacco mosaic virus (TMV) is evenly spread in acidic buffer solution, forms reaction solution, and stir；
2) dopamine hydrochloride and oxidant are added into reaction solution obtained by step 1), after being sufficiently stirred reaction, is added
100mM alkaline solution regulation pH value continues reaction 0.5~2 hour after being 7.4~8.5；
3) reaction solution obtained by step 2) is centrifuged, takes supernatant liquor, Surface coating is obtained after dialysing, being freeze-dried
The tobacco mosaic virus (TMV) nano wire of poly-dopamine.
Preferably, in step 1), the mass concentration of the reaction solution is 0.1~2.0mg/mL, and the mixing time is
0.5~4 hour；In step 2), the stirring reaction refers to be sufficiently stirred reaction under the conditions of 4~35 DEG C 2~8 hours；Step
3) in, the rotating speed of the centrifugation is 5000~10000rpm, and centrifugation time is 5~10min, and the dialysis refers to retention molecule
The bag filter measured as 10~1,000,000 dalton is dialysed 24~48 hours in pure water.
Preferably, in step 1), the acidic buffer solution is sodium acetate buffer solution, sodium citrate buffer or phosphorus
It is a kind of in hydrochlorate cushioning liquid (PBS)；The pH value of the acidic buffer solution is 4.5~6.8, and molar concentration is 50~100mM.
Preferably, in step 2), the oxidant is ammonium persulfate, one kind in hydrogen peroxide or oxygen；The alkalescence
Solution is sodium hydrate aqueous solution or the trishydroxymethylaminomethane aqueous solution (the Tris aqueous solution)；The dopamine hydrochloride and cigarette
The mass ratio of showy flowers of herbaceous plants mosaic virus is 0.2:1~2:1, the molar concentration rate of dopamine hydrochloride and oxidant is 1:2~1:4.
Preferably, in step 3), the tobacco mosaic virus (TMV) nano wire of the Surface coating poly-dopamine, its a diameter of 18~
40nm, length are 300nm~20 μm, and the thickness of poly-dopamine clad is 1~10nm.
Preferably, it is described to carry out functional modification using the poly-dopamine coated in gained nanowire surface, refer to table
The tobacco mosaic virus (TMV) nano wire that bread covers poly-dopamine is template, in alkaline buffer solution, the absorption of poly-dopamine clad
With reduction the solution containing gold ion, so as to realize growth of the gold nano grain on one-dimensional nano line surface；Or poly-dopamine
Clad is reacted with the molecule with bioactivity containing amino or sulfydryl, so as to realize functional molecular in 1-dimention nano
The modification on line surface, finally give described surface-functionalized tobacco mosaic virus (TMV) nanowire composite, specifically include as
Lower step：
A. the tobacco mosaic virus (TMV) nano wire of Surface coating poly-dopamine is evenly spread in alkaline buffer solution, formed
Reaction solution, and stir；
B. the solution containing gold ion is added into reaction solution described in step a, or is added containing amino or sulfydryl
Molecule with bioactivity, after reaction completely, by dialysing, being freeze-dried, obtain surface-functionalized tobacco mosaic virus (TMV)
Nanowire composite.
Preferably, in step a, the mass concentration of the reaction solution is 0.1~2.0mg/mL, and the mixing time is
0.5~2 hour；In step b, all referring to being reacted 2~24 hours under the conditions of 4 DEG C, the dialysis refers to use for the reaction
Molecular cut off is dialysed 24~48 hours for the bag filter of 10~1,000,000 dalton in pure water.
Preferably, in step a, the alkaline buffer solution is that (Tris-HCl delays trishydroxymethylaminomethane cushioning liquid
Fliud flushing), one kind in phosphate buffer solution (PBS) or sodium carbonate-bicarbonate buffer solution；The alkaline buffer solution pH value
For 7.8~8.5, molar concentration is 10~100mM.
Preferably, in step b, the solution containing gold ion is aqueous solution of chloraurate, its mass concentration is 0.01~
10mg/mL；The molecule with bioactivity containing amino or sulfydryl is cancer therapy drug doxorubicin hydrochloride (DOX) or tumour
Targeted molecular ring-type arginine-glycine-aspartic acid polypeptide (cRGD ring type polypeptides), the having containing amino or sulfydryl
The molecular mass concentration of bioactivity is 1~20mg/mL.
Beneficial effects of the present invention are as follows：
(1) solve the problems, such as that poly-dopamine coats in hydrophilic protein or surfaces of viral particles, by acidity
Dopamine under environment is positively charged, is readily adsorbed in the negatively charged protein surface in surface, while add ammonium persulfate etc.
Oxidant in-situ oxidizing-polymerizing dopamine generates poly-dopamine, so that hydrophilic protein or tobacco mosaic virus (TMV) surface are tight
One strata dopamine of close cladding, pH value is adjusted after the completion of reaction is basic again to neutral or alkalescent, further solidifies poly-dopamine
Cladding.This is clearly distinguishable from polymeric reaction condition of the dopamine described by art methods directly under alkaline environment.
(2) conductive polymer material is instead of with the poly-dopamine with biocompatibility, solves the biography such as polyaniline
Uniting high polymer material can not be biomedical sector apply the shortcomings that so that the tobacco that the surface poly-dopamine being prepared coats
There are the one-dimensional nano line of mosaic virus assembling potential biomedical applications to be worth.
(3) poly-dopamine make use of to adsorb the ability with reducing metal ion under alkaline environment, without adding volume
Outer reducing agent, you can in the tobacco mosaic virus (TMV) Surface Creation gold nano grain of poly-dopamine cladding, solve conventional method
Need to add reducing agent, it is necessary to carry out Tobacco mosaic virus lacking for the genetic modification of molecular biology, mutation and expression
Point, greatly simplify preparation method, shorten manufacturing cycle, and technique is simple, convenience and high-efficiency.
(4) energy that make use of poly-dopamine to be reacted under alkaline environment with the molecule containing amino or sulfydryl
Power, without extra chemical modification and modified method, you can in the tobacco mosaic virus (TMV) surface modification function of poly-dopamine cladding
Molecule.Such as cancer therapy drug doxorubicin hydrochloride DOX and tumor targeted molecular cRGD ring type polypeptides are passed through in the present invention simple
Mix just directly to modify and arrive one-dimensional tobacco mosaic virus (TMV) nanowire surface, solving conventional method needs extra chemistry
Reaction, intermediate steps more easily the shortcomings that making protein denaturation precipitation；Or need to divide Tobacco mosaic virus
The shortcomings that genetic modification, mutation and the expression of sub- biology are to produce reactive groups.
(5) the surface-functionalized tobacco mosaic virus (TMV) nanowire composite of present invention gained can be applied to living things catalysis,
The fields such as bio-imaging, cancer target probe and nano-medicament carrier.
Brief description of the drawings
The embodiment of the present invention is described in further detail below in conjunction with the accompanying drawings.
Fig. 1 shows a kind of surface-functionalized tobacco mosaic virus (TMV) nanowire composite preparation side of the embodiment of the present invention
The reaction process schematic diagram of method.
Fig. 2 shows the transmission electron microscope picture of the one-dimensional nano line composite prepared by the embodiment of the present invention 1.
Fig. 3 shows the scanning electron microscope (SEM) photograph of the one-dimensional nano line composite prepared by the embodiment of the present invention 1.
Fig. 4 shows the dynamic light scattering diagram of the one-dimensional nano line composite prepared by the embodiment of the present invention 1.
Fig. 5 shows ultraviolet-visible spectrogram and the calibration of the one-dimensional nano line composite prepared by the embodiment of the present invention 2
Curve (Fig. 5 (a)), and its ultraviolet absorption value at 314nm and 480nm change with time (Fig. 5 (b)).
Fig. 6 (a) (b) (c), which is shown respectively in the embodiment of the present invention 1, adds 5 μ L, 20 μ L, prepared by 100 μ L chlorauric acid solutions
The various sizes of gold nano grain composite of one-dimensional nano line area load transmission electron microscope picture.
Fig. 7 shows one-dimensional nano line surface modification cancer therapy drug doxorubicin hydrochloride prepared by the embodiment of the present invention 8 and swollen
The transmission electron microscope picture of knurl targeted molecular cRGD ring type polypeptide composites.
Embodiment
In order to illustrate more clearly of the present invention, the present invention is done further with reference to preferred embodiments and drawings
It is bright.Similar part is indicated with identical reference in accompanying drawing.It will be appreciated by those skilled in the art that institute is specific below
The content of description is illustrative and be not restrictive, and should not be limited the scope of the invention with this.
A kind of reaction of surface-functionalized tobacco mosaic virus (TMV) nanowire composite preparation method provided by the invention
Schematic flow sheet, as shown in Figure 1.
Embodiment 1
1mg tobacco mosaic virus (TMV) is distributed in the sodium acetate buffer solution 2mL for the 100mM that pH value is 5.5, is made into matter
The reaction solution that concentration is 0.5mg/mL is measured, after being stirred 2 hours under the conditions of 4 DEG C, 1mg dopamine is added into reaction solution
The ammonium persulfate of hydrochloride and 2.4mg, after reaction being sufficiently stirred under the conditions of 25 DEG C 4 hours, 100mM hydrogen-oxygen is added several times
Change sodium water solution 50uL, while monitored with the wide model pH test paper of 1-14 scopes, regulation reacting solution pH value continues to react after being 8.5
2 hours, reaction solution is then centrifuged into 10min under 9500rpm rotating speed, takes supernatant liquor, be 1,000,000 with molecular cut off
The bag filter of dalton is dialysed 48 hours in pure water, changes 1 st pure water for every eight hours, and finally freeze-drying obtains Surface coating and gathered
The tobacco mosaic virus (TMV) nano wire of dopamine.
The tobacco mosaic virus (TMV) nano wire of the Surface coating poly-dopamine of the above-mentioned preparations of 1mg is dispersed in 10mM pH
It is worth in the trishydroxymethylaminomethane cushioning liquid 2mL for 8.5, forms the reaction solution that mass concentration is 0.5mg/mL, stirring
0.5 hour, the μ L of 10mg/mL aqueous solution of chloraurate 5,20,100 are separately added into, reaction 24 is small after rapid mixing under the conditions of 4 DEG C
When, dialysed with molecular cut off for the bag filter of 1,000,000 dalton in pure water 48 hours, change 1 st pure water for every eight hours, finally
It is freeze-dried to obtain one-dimensional tobacco mosaic virus (TMV) nano wire composite wood of the different size of gold nano grain in area load
Material.
The transmission electron microscope picture of the tobacco mosaic virus (TMV) nano wire of Surface coating poly-dopamine prepared by the present embodiment and sweep
Electron microscope is retouched, as shown in Figures 2 and 3, it was demonstrated that a diameter of 22~26nm of this one-dimensional tobacco mosaic virus (TMV) nano wire, it is long
Spend for 2~20 μm, the thickness of poly-dopamine clad is 4~8nm.This one-dimensional tobacco mosaic disease prepared by the present embodiment
The dynamic light scattering diagram of malicious nano wire, as shown in figure 4, the hydration radius for having neither part nor lot in the tobacco mosaic virus (TMV) to form nano wire is
89.4nm, the hydration radius for the tobacco mosaic virus (TMV) for participating in forming nano wire is 721.8nm, considerably beyond tobacco mosaic virus (TMV)
The hydration radius of itself, it is one-dimensional nano wire package assembly in the solution to further demonstrate it.
The one-dimensional tobacco mosaic virus (TMV) nanometer of different size of gold nano grain prepared by the present embodiment in area load
The transmission electron microscope picture of line composite is as shown in fig. 6, the gold nano grain of generation is equal on the surface of one-dimensional nano line composite
Even distribution.Fig. 6 (a) is adds the reaction of 5 μ L gold chlorides, and the gold nano grain diameter of generation is in 2~5nm；Fig. 6 (b) is 20 μ of addition
L gold chlorides react, and the gold nano grain diameter of generation is in 10~30nm；Fig. 6 (c) reacts to add 100 μ L gold chlorides, generation
Gold nano grain diameter is in 200~300nm.This example demonstrates that the Jenner that the addition of gold chloride can be generated with significant impact
Rice grain size.
Embodiment 2
1mg tobacco mosaic virus (TMV) is distributed in the sodium acetate buffer solution 2mL for the 100mM that pH value is 5.5, is made into matter
The reaction solution that concentration is 0.5mg/mL is measured, after being stirred 2 hours under the conditions of 4 DEG C, 1mg dopamine is added into reaction solution
The ammonium persulfate of hydrochloride and 2.4mg, be sufficiently stirred under the conditions of 25 DEG C reaction 24 hours, temporally put sampling and testing it is ultraviolet-
Visible spectrum absorption value.Then reaction solution is centrifuged into 10min under 9500rpm rotating speed, takes supernatant liquor, with retention molecule
Measure and dialysed for the bag filter of 1,000,000 dalton in pure water 48 hours, change 1 st pure water for every eight hours, be freeze-dried.
The tobacco mosaic virus (TMV) nano wire of Surface coating poly-dopamine prepared by the present embodiment it is ultraviolet with time sampling
Shown in visible absorption spectra figure such as Fig. 5 (a), its ultraviolet absorption value at 314nm and 480nm, which is plotted against time, to be linearly increased
Shown in trend such as Fig. 5 (b), the reaction for indicating dopamine oxidation polymerization in acid condition is gradually increased.
Embodiment 3
1mg tobacco mosaic virus (TMV) is distributed in the sodium acetate buffer solution 2mL for the 100mM that pH value is 5.5, is made into matter
The reaction solution that concentration is 0.5mg/mL is measured, after being stirred 2 hours under the conditions of 4 DEG C, 0.2 is separately added into reaction solution,
0.4,0.6,0.8,1.0,1.2,1.4,1.6,1.8,2.0mg dopamine hydrochloride and molar concentration rate is 1:2 persulfuric acid
0.48~4.8mg of ammonium, after reaction being sufficiently stirred under the conditions of 25 DEG C 4 hours, 100mM sodium hydrate aqueous solution is added several times
50uL, while monitored with the wide model pH test paper of 1-14 scopes, regulation reacting solution pH value continues reaction 2 hours after being 8.5, then
Reaction solution is centrifuged into 10min under 9500rpm rotating speed, takes supernatant liquor, is the saturating of 1,000,000 dalton with molecular cut off
Analysis bag is dialysed 48 hours in pure water, changes 1 st pure water for every eight hours, and finally freeze-drying, which obtains Surface coating, has different-thickness to gather
The tobacco mosaic virus (TMV) nano wire of dopamine.
The present embodiment carries out the cladding reality of tobacco mosaic virus (TMV) nano wire by adding the dopamine hydrochloride of different proportion
Test, the mass ratio for showing the dopamine hydrochloride and tobacco mosaic virus (TMV) added is 1:It is optimal when 1.
Embodiment 4
By 1mg tobacco mosaic virus (TMV) be distributed to respectively pH value be 4.5,5.5,6.0 50mM sodium acetate buffer solution,
The trishydroxymethylaminomethane cushioning liquid for the 50mM that pH value is 6.8,7.4 50mM phosphate buffer solution, pH value is 8.5
In (Tris-HCl buffer solutions), the reaction solution that mass concentration is 0.5mg/mL is made into, after being stirred 2 hours under the conditions of 4 DEG C, to
1mg dopamine hydrochloride and 2.4mg ammonium persulfate are added in reaction solution, it is small that reaction 4 is sufficiently stirred under the conditions of 25 DEG C
Shi Hou, 100mM sodium hydrate aqueous solution 50uL is added several times, while monitored with the wide model pH test paper of 1-14 scopes, regulation
Reacting solution pH value continues reaction 2 hours after being 8.5, and reaction solution then is centrifuged into 10min under 9500rpm rotating speed, taken
Supernatant liquor, dialysed 48 hours, changed for every eight hours once pure in pure water for the bag filter of 1,000,000 dalton with molecular cut off
Water, freeze-drying.
The cladding that the present embodiment carries out tobacco mosaic virus (TMV) nano wire by adding the cushioning liquid of different pH value is tested, table
Bright solution ph is higher, and the reaction speed of dopamine polymerization is faster, but covered effect is also poorer simultaneously.In the second that pH value is 5.5
The tobacco mosaic virus (TMV) nano wire prepared in sour sodium cushioning liquid is optimal.
Embodiment 5
1mg tobacco mosaic virus (TMV) is distributed in the sodium acetate buffer solution 2mL for the 100mM that pH value is 5.5, is made into matter
The reaction solution that concentration is 0.5mg/mL is measured, after being stirred 2 hours under the conditions of 4 DEG C, the more of 1mg are separately added into reaction solution
Bar amine hydrochlorate and 2.4,3.6,4.8mg (molar concentration rates 1:2,1:3,1:4) ammonium persulfate, under the conditions of 25 DEG C fully
After stirring reaction 4 hours, 100mM sodium hydrate aqueous solution 50uL is added several times, while tried with the wide model pH of 1-14 scopes
Paper monitor, regulation reacting solution pH value be 8.5 after continue reaction 2 hours, then by reaction solution under 9500rpm rotating speed from
Heart 10min, takes supernatant liquor, is dialysed with molecular cut off for the bag filter of 1,000,000 dalton in pure water 48 hours, every 8 is small
When change 1 st pure water, be freeze-dried.
The present embodiment carries out the cladding of tobacco mosaic virus (TMV) nano wire by adding the ammonium persulfate oxidizing agent of different proportion
Experiment, shows that excessive oxidant can accelerate the polymerization rate of dopamine but covered effect is poor simultaneously.Dopamine hydrochloric acid
The molar concentration rate of salt and ammonium persulfate is 1:It is optimal when 2.
Embodiment 6
1mg tobacco mosaic virus (TMV) is distributed in the sodium citrate buffer 2mL for the 50mM that pH value is 4.5, be made into
Mass concentration is 0.1mg/mL reaction solution, and after being stirred 4 hours under the conditions of 4 DEG C, the more of 0.2mg are added into reaction solution
Bar amine hydrochlorate, and bubble oxygen is continually fed into, after reaction being sufficiently stirred under the conditions of 35 DEG C 8 hours, 100mM is added several times
Trishydroxymethylaminomethane aqueous solution 50uL, while monitored with the wide model pH test paper of 1-14 scopes, adjust reacting solution pH value
To continue reaction 0.5 hour after 7.4, reaction solution is then centrifuged into 5min under 10000rpm rotating speed, takes supernatant liquor, used
Molecular cut off is dialysed 24 hours for the bag filter of 100,000 dalton in pure water, changes 1 st pure water for every eight hours, and finally freezing is dry
The dry tobacco mosaic virus (TMV) nano wire for obtaining Surface coating poly-dopamine.
The tobacco mosaic virus (TMV) nano wire of the Surface coating poly-dopamine of the above-mentioned preparations of 1mg is dispersed in 100mM's
PH value is in 7.8 phosphate buffer solution, forms the reaction solution that mass concentration is 2.0mg/mL, stirs 2 hours, adds matter
The cancer therapy drug doxorubicin hydrochloride (DOX) that concentration is 20mg/mL is measured, is turned after being reacted 2 hours under the conditions of 4 DEG C with 9500rpm
Speed centrifugation 5min, takes supernatant liquor, is dialysed with molecular cut off for the bag filter of 100,000 dalton in pure water 24 hours, and every 8
Hour changes a water, and finally freeze-drying obtains surface modification cancer therapy drug doxorubicin hydrochloride DOX one-dimensional tobacco mosaic virus (TMV)
Nanowire composite.
Embodiment 7
1mg tobacco mosaic virus (TMV) is distributed in the phosphate buffer solution 2mL for the 100mM that pH value is 6.8, is made into matter
The reaction solution that concentration is 2.0mg/mL is measured, after being stirred 0.5 hour under the conditions of 4 DEG C, 2mg DOPA is added into reaction solution
The molar concentration rate of amine hydrochlorate and ammonium persulfate, dopamine and ammonium persulfate is 1:4, reaction 2 is sufficiently stirred under the conditions of 4 DEG C
After hour, 100mM sodium hydrate aqueous solution 50uL is added several times, while is monitored with the wide model pH test paper of 1-14 scopes, is adjusted
Section reacting solution pH value continues reaction 2 hours after being 8.5, and reaction solution then is centrifuged into 10min under 5000rpm rotating speed,
Supernatant liquor is taken, is dialysed 48 hours, changed for every eight hours once in pure water for the bag filter of 1,000,000 dalton with molecular cut off
Pure water, finally freeze-drying obtain the tobacco mosaic virus (TMV) nano wire of Surface coating poly-dopamine.
The tobacco mosaic virus (TMV) nano wire of the Surface coating poly-dopamine of the above-mentioned preparations of 1mg is dispersed in 10mM pH
It is worth in the trishydroxymethylaminomethane cushioning liquid for 8.5, forms the reaction solution that mass concentration is 0.5mg/mL, add 10mg
Tumor targeted molecular ring-type arginine-glycine-aspartic acid polypeptide, with 9500rpm's after being reacted 2 hours under the conditions of 4 DEG C
Rotating speed centrifuges 5min, takes supernatant liquor, is dialysed with molecular cut off for the bag filter of 1,000,000 dalton in pure water 24 hours,
1 st pure water is changed for every eight hours, and finally freeze-drying obtains the 1-dimention nano of surface modification tumor targeted molecular cRGD ring type polypeptides
Line composite.
Embodiment 8
The tobacco mosaic virus (TMV) nano wire of Surface coating poly-dopamine prepared by 1mg above-described embodiments 1 is dispersed in
10mM pH value is in 8.5 trishydroxymethylaminomethane cushioning liquid, forms the reaction solution that mass concentration is 0.5mg/mL,
5mg cancer therapy drugs doxorubicin hydrochloride (DOX) and 5mg tumor targeted molecular cRGD ring type polypeptides are sequentially added, it is anti-under the conditions of 4 DEG C
After answering 4 hours, 5min is centrifuged with 9500rpm rotating speed, takes supernatant liquor, with the dialysis that molecular cut off is 1,000,000 dalton
Bag dialyse 24 hours in pure water, changes 1 st pure water for every eight hours, be finally freeze-dried obtain area load cancer therapy drug hydrochloric acid Ah
The one-dimensional nano line composite of difunctionalization of mycin (DOX) and tumor targeted molecular cRGD ring type polypeptides modification.
The one-dimensional nano line composite of cancer therapy drug and the modification of targeted molecular difunctionalization prepared by the present embodiment
Transmission electron microscope picture is as shown in fig. 7, the composite modified by difunctionalization can also keep its one-dimensional nano wire package assembly.
Obviously, the above embodiment of the present invention is only intended to clearly illustrate example of the present invention, and is not pair
The restriction of embodiments of the present invention, for those of ordinary skill in the field, may be used also on the basis of the above description
To make other changes in different forms, all embodiments can not be exhaustive here, it is every to belong to this hair
Row of the obvious changes or variations that bright technical scheme is extended out still in protection scope of the present invention.
Claims (8)
- A kind of 1. preparation method of surface-functionalized tobacco mosaic virus (TMV) nanowire composite, it is characterised in that：Including such as Lower step：The assembling of the tobacco mosaic virus (TMV) nano wire of Surface coating poly-dopamine is first carried out, recycles gained nanowire surface The poly-dopamine of upper cladding carries out functional modification, obtains surface-functionalized tobacco mosaic virus (TMV) nanowire composite；The number of assembling steps of the tobacco mosaic virus (TMV) nano wire of the Surface coating poly-dopamine is as follows：1）Tobacco mosaic virus (TMV) is evenly spread in acidic buffer solution, forms reaction solution, and stir；2）To step 1）Dopamine hydrochloride and oxidant are added in gained reaction solution, after being sufficiently stirred reaction, adds 100mM Alkaline solution regulation pH value be 7.4~8.5 after continue reaction 0.5~2 hour；3）By step 2）Gained reaction solution is centrifuged, and takes supernatant liquor, and it is poly- more that Surface coating is obtained after dialysing, being freeze-dried The tobacco mosaic virus (TMV) nano wire of bar amine；It is described as follows using the poly-dopamine progress functional modification step coated in gained nanowire surface：A. the tobacco mosaic virus (TMV) nano wire of Surface coating poly-dopamine is evenly spread in alkaline buffer solution, formed anti- Solution is answered, and is stirred；B. the solution containing gold ion is added into reaction solution described in step a, or adds having containing amino or sulfydryl The molecule of bioactivity;After reaction completely, by dialysing, being freeze-dried, surface-functionalized tobacco mosaic virus (TMV) nanometer is obtained Line composite.
- 2. preparation method according to claim 1, it is characterised in that：Step 1）In, the mass concentration of the reaction solution For 0.1~2.0mg/mL, the mixing time is 0.5~4 hour；Step 2）In, the stirring reaction refers in 4~35 DEG C of bars Reaction is sufficiently stirred under part 2~8 hours；Step 3）In, the rotating speed of the centrifugation is 5000~10000rpm, centrifugation time 5 ~10min, the dialysis refer to be dialysed in pure water for the bag filter of 10~1,000,000 dalton with molecular cut off 24~48 small When.
- 3. preparation method according to claim 1, it is characterised in that：Step 1）In, the acidic buffer solution is acetic acid It is a kind of in sodium cushioning liquid, sodium citrate buffer or phosphate buffer solution；The pH value of the acidic buffer solution is 4.5 ~6.8, molar concentration is 50~100mM.
- 4. preparation method according to claim 1, it is characterised in that：Step 2）In, the oxidant is ammonium persulfate, mistake One kind in hydrogen oxide or oxygen；The alkaline solution is sodium hydrate aqueous solution or the trishydroxymethylaminomethane aqueous solution；Institute The mass ratio for stating dopamine hydrochloride and tobacco mosaic virus (TMV) is 0.2:1~2:1, dopamine hydrochloride and oxidant it is mole dense Degree is than being 1:2~1:4.
- 5. preparation method according to claim 1, it is characterised in that：Step 3）In, the Surface coating poly-dopamine Tobacco mosaic virus (TMV) nano wire, its a diameter of 18~40nm, length are 300nm~20 μm, and the thickness of poly-dopamine clad is 1 ~10nm.
- 6. preparation method according to claim 1, it is characterised in that：In step a, the mass concentration of the reaction solution is 0.1~2.0mg/mL, the mixing time are 0.5~2 hour；In step b, the reaction is all referring to anti-under the conditions of 4 DEG C To answer 2~24 hours, the dialysis refers to dialyse 24 in pure water with molecular cut off for the bag filter of 10~1,000,000 dalton~ 48 hours.
- 7. preparation method according to claim 1, it is characterised in that：In step a, the alkaline buffer solution is three hydroxyl first One kind in base aminomethane buffer solution, phosphate buffer solution or sodium carbonate-bicarbonate buffer solution；The alkaline buffer Solution ph is 7.8~8.5, and molar concentration is 10~100mM.
- 8. preparation method according to claim 1, it is characterised in that：In step b, the solution containing gold ion is chlorine The auric acid aqueous solution, its mass concentration are 0.01~10mg/mL；The molecule with bioactivity containing amino or sulfydryl is Doxorubicin hydrochloride or ring-type arginine-glycine-aspartic acid polypeptide；It is described that there is bioactivity containing amino or sulfydryl Molecular mass concentration is 1~20mg/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510696672.5A CN105327354B (en) | 2015-10-23 | 2015-10-23 | A kind of preparation method of surface-functionalized tobacco mosaic virus (TMV) nanowire composite |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510696672.5A CN105327354B (en) | 2015-10-23 | 2015-10-23 | A kind of preparation method of surface-functionalized tobacco mosaic virus (TMV) nanowire composite |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105327354A CN105327354A (en) | 2016-02-17 |
| CN105327354B true CN105327354B (en) | 2018-03-27 |
Family
ID=55278255
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510696672.5A Active CN105327354B (en) | 2015-10-23 | 2015-10-23 | A kind of preparation method of surface-functionalized tobacco mosaic virus (TMV) nanowire composite |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105327354B (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107312179B (en) * | 2016-04-26 | 2020-08-25 | 中国科学院理化技术研究所 | Preparation method of tobacco mosaic virus-poly (N-isopropylacrylamide) block copolymer Pickering emulsion |
| CN107998101B (en) * | 2017-12-05 | 2020-10-27 | 华中农业大学 | Method for embedding hydrophobic drug |
| CN108339125B (en) * | 2018-03-23 | 2021-06-04 | 温州生物材料与工程研究所 | Targeted drug-loaded nano micelle and preparation method and application thereof |
| CN108640980B (en) * | 2018-05-24 | 2020-12-11 | 中国科学院理化技术研究所 | Protein assembly and preparation method thereof |
| CN108938596B (en) * | 2018-08-03 | 2020-07-28 | 宁夏医科大学 | CPPs/NGR modified drug-loaded nano gold rod and preparation method and application thereof |
| CN111087472A (en) * | 2018-10-23 | 2020-05-01 | 中国海洋大学 | Preparation method of biological active diatom biomineralization silicon |
| US11529430B2 (en) * | 2019-12-20 | 2022-12-20 | Case Western Reserve University | Polydopamine decorated tobacco mosaic theranostic virus nanoparticles |
| CN111349149A (en) * | 2020-03-17 | 2020-06-30 | 中国科学院苏州纳米技术与纳米仿生研究所 | Tobacco mosaic virus capsid protein mutant and application thereof |
| CN111679021B (en) * | 2020-07-06 | 2023-09-01 | 大连工业大学 | Method for detecting heterocyclic amine in solid sample |
| CN113385046B (en) * | 2021-06-24 | 2022-05-10 | 中国矿业大学 | Preparation method and application of sponge-based filter membrane adsorbing material for mercury adsorption removal |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102049226A (en) * | 2010-11-16 | 2011-05-11 | 中国烟草总公司郑州烟草研究院 | Method for preparing TMV template-based nano-gold material |
| CN103706800A (en) * | 2014-01-16 | 2014-04-09 | 国家烟草质量监督检验中心 | Method for growing nanogold on gene-modified tobacco mosaic virus nuclecapsid protein template |
-
2015
- 2015-10-23 CN CN201510696672.5A patent/CN105327354B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102049226A (en) * | 2010-11-16 | 2011-05-11 | 中国烟草总公司郑州烟草研究院 | Method for preparing TMV template-based nano-gold material |
| CN103706800A (en) * | 2014-01-16 | 2014-04-09 | 国家烟草质量监督检验中心 | Method for growing nanogold on gene-modified tobacco mosaic virus nuclecapsid protein template |
Non-Patent Citations (4)
| Title |
|---|
| "Assembly of Tobacco Mosaic Virus into Fibrous and Macroscopic Bundled Arrays Mediated by Surface Aniline Polymerization";Zhongwei Niu et al.;《Langmuir》;20070503;第23卷(第12期);第6719-6724页 * |
| "Mussel-Inspired Polydopamine Coating on Tobacco Mosaic Virus: One-Dimensional Hybrid Nanofibers for Gold Nanoparticle Growth";Quan Zhou et al.;《Langmuir》;20170829;第33卷(第38期);第9866-9872页 * |
| "Self-Assembly of Tobacco Mosaic Virus at Oil/Water Interfaces";Jinbo He et al.;《Langmuir》;20090401;第25卷(第9期);第4979-4987页 * |
| "一维棒状烟草花叶病毒的自组装及其应用";刘朋等;《化学进展》;20151015;第27卷(第10期);第1425-1434页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105327354A (en) | 2016-02-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105327354B (en) | A kind of preparation method of surface-functionalized tobacco mosaic virus (TMV) nanowire composite | |
| Steinmetz et al. | Utilisation of plant viruses in bionanotechnology | |
| Dixit et al. | Quantum dot encapsulation in viral capsids | |
| Mercante et al. | Nanofibers interfaces for biosensing: Design and applications | |
| Iriarte‑Mesa et al. | Gold, silver and iron oxide nanoparticles: synthesis and bionanoconjugation strategies aiming to electrochemical applications | |
| Mamidi et al. | Carbonaceous nanomaterials incorporated biomaterials: The present and future of the flourishing field | |
| Qiu et al. | Synthesis and characterization of ferrocene modified Fe3O4@ Au magnetic nanoparticles and its application | |
| Yoon et al. | Polymeric supramolecular assemblies based on multivalent ionic interactions for biomedical applications | |
| Sarkar et al. | Protein nanocomposites: Special inferences to lysozyme based nanomaterials | |
| CN103524750B (en) | The preparation method of Polyethylene glycol chitosan self-assembled nanometer grain | |
| Hosta et al. | Conjugation of Kahalalide F with gold nanoparticles to enhance in vitro antitumoral activity | |
| Bharathala et al. | Biomedical applications of nanoparticles | |
| Elancheziyan et al. | Covalent immobilization and enhanced electrical wiring of hemoglobin using gold nanoparticles encapsulated PAMAM dendrimer for electrochemical sensing of hydrogen peroxide | |
| Zhang et al. | Fabrication of degradable lemon-like porous silica nanospheres for pH/redox-responsive drug release | |
| Li et al. | Surface functionalization of nanomaterials by aryl diazonium salts for biomedical sciences | |
| Singh et al. | Preparation of self-activated fluorescence mesoporous silica hollow nanoellipsoids for theranostics | |
| Baig et al. | Synthesis of ligand functionalized ErbB-3 targeted novel DNA nano-threads loaded with the low dose of doxorubicin for efficient in vitro evaluation of the resistant anti-cancer activity | |
| Xu et al. | Real-time cell analysis of the cytotoxicity of a pH-responsive drug-delivery matrix based on mesoporous silica materials functionalized with ferrocenecarboxylic acid | |
| Hosseinkhani | Biomedical Engineering: Materials, Technology, and Applications | |
| Wang et al. | Functionalizing DNA nanostructures with natural cationic amino acids | |
| Buniyamin et al. | Nanotechnology applications in biomedical systems | |
| CN103990138B (en) | Layer-by-layer assembled nanogold composite drug delivery carrier system, preparation method and application thereof | |
| Baig et al. | Targeting folate receptors (α1) to internalize the bleomycin loaded DNA-nanotubes into prostate cancer xenograft CWR22R cells | |
| Baruah et al. | Guar gum-based hydrogel and hydrogel nanocomposites for biomedical applications | |
| Kang et al. | Disposable amperometric biosensor based on nanostructured bacteriophages for glucose detection |
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 |