CN105079806A - Carbon nanomaterial directly modified by polypeptide as well as preparation method and application thereof - Google Patents
Carbon nanomaterial directly modified by polypeptide as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN105079806A CN105079806A CN201410201129.9A CN201410201129A CN105079806A CN 105079806 A CN105079806 A CN 105079806A CN 201410201129 A CN201410201129 A CN 201410201129A CN 105079806 A CN105079806 A CN 105079806A
- Authority
- CN
- China
- Prior art keywords
- carbon nanomaterial
- polypeptide
- carbon
- peptide
- oxidation state
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Carbon And Carbon Compounds (AREA)
Abstract
The invention provides a carbon nanomaterial directly modified by polypeptide. The carbon nanomaterial directly modified by polypeptide comprises polypeptide molecules and a carbon nanomaterial body, wherein the polypeptide molecules are connected with the carbon nanomaterial body through covalent bonds, the carbon nanomaterial directly modified by polypeptide is a product prepared by reducing the oxidation-state nanomaterial by adopting polypeptide molecules, the polypeptide molecules contain reductive amino acid, and the oxidation-state nanomaterial contains surface active groups which are carboxyls, epoxy groups or hydroxyls. The carbon nanomaterial directly modified by polypeptide has good stability, and the activity of polypeptide and the dispersibility of the carbon nanomaterial are not obviously changed. The invention further provides a preparation method and an application thereof the carbon nanomaterial directly modified by polypeptide.
Description
Technical field
The present invention relates to production of polysilicon equipment technical field, be specifically related to carbon nanomaterial that a peptide species directly modifies and its preparation method and application.
Background technology
Carbon nanomaterial has special physicochemical property, carbon nanomaterial is (as CNT, Graphene, carbon nano-particle etc.) there is very strong absorbing properties, its surface can be used in the localized heat imaging of tumor after biological functional is modified, photo-thermal therapy, the aspect such as photoacoustic imaging and fluorescence sense.Bio-modification at present based on carbon nanomaterial mainly contains physical absorption and covalent coupling two kinds of technology.
Together with physical absorption is adsorbed onto by π-π interaction self assembly between the carbon nanomaterial that utilizes the molecule (as organic dyestuff probe) with phenyl ring conjugation abundant with pi-electron; This method is simple and convenient, but because this kind of assembling is not covalent coupling, its stability is easily subject to the pH of solution, ionic strength, the impact of the factors such as salinity.
Covalent coupling is technology conventional to carbon nanomaterial bio-modification at present, first chemical treatment (as: oxidation) is carried out by carbon nanomaterial, its surface is allowed to have active group, then the coupling technology (as: EDC activation) of some classics is being passed through by the surface of biomacromolecule (albumen, nucleic acid and polypeptide etc.) covalent coupling to carbon nanomaterial; This method of modifying good stability, but Bioconjugation process can affect the dispersibility of carbon nanomaterial and the activity of biomolecule.
Therefore, be necessary to provide the stability of material after a kind of bio-modification better, and little on the dispersibility impact of carbon nanomaterial, to the little bio-modification carbon nanomaterial of the activity influence of biomolecule method.
Summary of the invention
For overcoming the defect of above-mentioned prior art, first aspect present invention provides the carbon nanomaterial that a peptide species is directly modified, the carbon nanomaterial that described polypeptide is directly modified has good stability, and the activity of polypeptide and the dispersibility of carbon nanomaterial all obviously do not change.Second aspect present invention provides the preparation method of the carbon nanomaterial that a peptide species is directly modified, described preparation method is by the method for a step reduction, the carbon nanomaterial adopting the polypeptide with reducing property surface to be in oxidation state reduces, and directly polypeptide is coupled to the surface of carbon nanomaterial.Third aspect present invention provides the application of the carbon nanomaterial that a peptide species is directly modified.
First aspect, the invention provides the carbon nanomaterial that a peptide species is directly modified, the carbon nanomaterial that described polypeptide is directly modified comprises peptide molecule and carbon nanomaterial, wherein, described peptide molecule is connected by covalent bond with carbon nanomaterial, the product of carbon nanomaterial obtained by the carbon nanomaterial of employing peptide molecule reduction-oxidation state that described polypeptide is directly modified, wherein, described peptide molecule contains reproducibility aminoacid, the carbon nanomaterial of described oxidation state contains surface active groups, the surface active groups of the carbon nanomaterial of described oxidation state is carboxyl, epoxy radicals or hydroxyl.
Preferably, the carbon nanomaterial of described oxidation state is the carbonoxide nano material of zero dimension.
Under this optimum condition, the carbonoxide nano material of described zero dimension does not have spatial orientation due to it, the impact not by space structure when peptide molecule and its effect.So peptide molecule is random uniform in the distribution on the surface of the carbonoxide nano material of described zero dimension, thus makes the carbon nanomaterial after modification have more stable character.
In addition, compared with multidimensional material with carbon element, the carbonoxide nano material size of zero dimension is more homogeneous, particle diameter only has tens nanometers, with peptide molecule, there is better size compatibility, because size is relatively less, it will have better cell-penetrating ability, will have more obvious advantage in biologic applications.
Further preferably, the carbonoxide nano material of described zero dimension is fullerene.
Preferably, the carbon nanomaterial of described oxidation state is carbonoxide nano-particle.
Further preferably, carbonoxide nano-particle of the present invention is prepared by following step:
1) wax candle ash is collected;
2) take out the wax candle ash of certain mass, add equal-volume DMF solvent and concentrated nitric acid, at a certain temperature back flow reaction a period of time;
3) step (2) gained reactant liquor is separated by gradient centrifugation, collects pitchy precipitation, and with after the water dissolution precipitation of certain volume, ultrasonic disperse process, namely obtains the solution containing described oxidized form carbon nano-particle.
Still more preferably, above the described wax candle ash flame that is wax candle, not clean-burning black smog is attached to the atrament on ceramic tile.
Still more preferably, the step that described gradient centrifugation is separated is:
A) slow-speed of revolution 6000 revs/min, centrifugal 5 minutes, by the removing of large-sized granule precipitation, collects upper strata black liquor and ultrasonic disperse process 2 hours;
B) with diluted sodium hydroxide solution, the pH value of step (a) gained black liquor is adjusted to neutrality, then by liquid through 9000 revs/min, centrifugal 5 minutes, further by granule precipitate and separate relatively large for size, collect upper strata pitchy transparency liquid;
C) by step (b) gained pitchy transparency liquid with 13000 revs/min, centrifugal 10 minutes, collect pitchy precipitation, and precipitate and standardize solution with the water dissolution of certain volume, ultrasonic disperse process 30 minutes, namely obtains the solution containing described oxidized form carbon nano-particle.
Still more preferably, above the described wax candle ash flame that is wax candle, not clean-burning black smog is attached to the atrament on ceramic tile.
Under this optimum condition, can obtain described carbonoxide nano-particle, described carbonoxide nano-particle comprises the carbonoxide nano material of zero dimension.
Preferably, the carbon nanomaterial of described oxidation state is graphene oxide.
Preferably, the carbon nanomaterial of described oxidation state is oxide/carbon nanometer tube.
Preferably, described reproducibility aminoacid is TYR, cysteine, histidine or tryptophan.
Preferably, the reproducibility aminoacid that the present invention adopts includes but not limited to aromatic amino acid.
Preferably, described is containing 5 ~ 40 amino acid whose polypeptide containing the amino acid whose peptide molecule of reproducibility.
Preferably, described is the polypeptide with site targeting containing the amino acid whose peptide molecule of reproducibility.
Preferably, described is neurotoxic peptide, RGD cyclic peptide or antibacterial polypeptide containing the amino acid whose peptide molecule of reproducibility.
Further preferably, the aminoacid sequence of described neurotoxic peptide comprises the aminoacid sequence as shown in SEQIDNO:1.
Still more preferably, the aminoacid sequence of described neurotoxic peptide is for such as shown in SEQIDNO:1.
Particularly, described RGD cyclic peptide is the circular polypeptides containing arginyl-glycyl-aspartic acid tripeptides, wherein, and described arginyl-glycyl-aspartic acid and Arg-Gly-Asp (RGD).
Further preferably, described RGD cyclic peptide is c (RGDfK).
Particularly, No. PubChemCommpoundCID of described c (RGDfK) is 10196873.
Further preferably, described antibacterial polypeptide is nisin.
Still more preferably, No. PubChemCommpoundCID of described nisin is 16219761.
Preferably, the size of carbon nanomaterial that polypeptide is directly modified is 30 ~ 60nm.
The carbon nanomaterial directly modified of the polypeptide that the present invention passes through utilizes the polypeptide with reducing property to reduce surface and is in the carbon nanomaterial of oxidation state, and peptide molecule is coupled to carbon nanomaterial surface by a direct step.In this coupling process, form covalent bond, compare physical absorption between the active group on the carbon nanomaterial surface of described reproducibility aminoacid and described oxidation state by redox reaction, the stability of the peptide modified carbon nanomaterial of gained is better.
In addition, compare traditional covalent coupling technology, the EDC that the present invention requires no biomolecule activates, also need not carry out the modifications such as PEG to carbon nanomaterial, polypeptid covalence can be coupled to the surface of carbon nanomaterial, the biological activity of the reasonable maintenance peptide molecule of energy of the peptide modified carbon nanomaterial of gained, and greatly reduce bio-modification process to the impact such as size, surface nature, water solublity of material with carbon element, thus reduce bio-modification process to the impact of carbon nanomaterial dispersibility; Therefore, the dispersibility of peptide modified carbon nanomaterial provided by the invention and water solublity better.
Second aspect, the invention provides the preparation method of the carbon nanomaterial that a peptide species is directly modified, comprises the steps:
1) carbon nanomaterial that peptide molecule that mass ratio is 1 ~ 4:1 ~ 20 and surface are in oxidation state is got, wherein, described peptide molecule contains reproducibility aminoacid, described surface be in oxidation state containing surface active groups, the surface active groups of the carbon nanomaterial of described oxidation state is carboxyl, epoxy radicals or hydroxyl;
2) carbon nanomaterial described surface being in oxidation state adds in buffer, after supersound process, add described peptide molecule and mixing obtain mixture;
3) gained mixture stirring reaction 8 ~ 24 hours at normal temperatures, reaction products therefrom ultra-filtration centrifuge tube adds buffer and supersound process, obtains the carbon nanomaterial that described polypeptide is directly modified after carrying out centrifugal, washing.
Preferably, in described step (1), the carbon nanomaterial of described oxidation state is the carbonoxide nano material of zero dimension.
Further preferably, the carbonoxide nano material of described zero dimension is fullerene.
Preferably, in described step (1), the carbon nanomaterial of described oxidation state is carbonoxide nano-particle.
Further preferably, carbonoxide nano-particle of the present invention is prepared by following step:
1) wax candle ash is collected;
2) take out the wax candle ash of certain mass, add equal-volume DMF solvent and concentrated nitric acid, at a certain temperature back flow reaction a period of time;
3) step (2) gained reactant liquor is separated by gradient centrifugation, collects pitchy precipitation, and with after the water dissolution precipitation of certain volume, ultrasonic disperse process, namely obtains the solution containing described oxidized form carbon nano-particle.
Still more preferably, above the described wax candle ash flame that is wax candle, not clean-burning black smog is attached to the atrament on ceramic tile.
Still more preferably, the step that described gradient centrifugation is separated is:
A) slow-speed of revolution 6000 revs/min, centrifugal 5 minutes, by the removing of large-sized granule precipitation, collects upper strata black liquor and ultrasonic disperse process 2 hours;
B) with diluted sodium hydroxide solution, the pH value of step (a) gained black liquor is adjusted to neutrality, then by liquid through 9000 revs/min, centrifugal 5 minutes, further by granule precipitate and separate relatively large for size, collect upper strata pitchy transparency liquid;
C) by step (b) gained pitchy transparency liquid with 13000 revs/min, centrifugal 10 minutes, collect pitchy precipitation, and precipitate and standardize solution with the water dissolution of certain volume, ultrasonic disperse process 30 minutes, namely obtains the solution containing described oxidized form carbon nano-particle.
Still more preferably, above the described wax candle ash flame that is wax candle, not clean-burning black smog is attached to the atrament on ceramic tile.
Under this optimum condition, can obtain described carbonoxide nano-particle, described carbonoxide nano-particle comprises the carbonoxide nano material of zero dimension.
Preferably, in described step (1), the carbon nanomaterial of described oxidation state is graphene oxide.
Preferably, in described step (1), the carbon nanomaterial of described oxidation state is oxide/carbon nanometer tube.
Preferably, in described step (1), described reproducibility aminoacid is TYR, cysteine, histidine or tryptophan.
Preferably, in described step (1), the reproducibility aminoacid that the present invention adopts includes but not limited to aromatic amino acid.
Preferably, in described step (1), described is containing 5 ~ 40 amino acid whose polypeptide containing the amino acid whose peptide molecule of reproducibility.
Preferably, in described step (1), described is the polypeptide with site targeting containing the amino acid whose peptide molecule of reproducibility.
Preferably, described is neurotoxic peptide, RGD cyclic peptide or antibacterial polypeptide containing the amino acid whose peptide molecule of reproducibility.
Further preferably, the aminoacid sequence of described neurotoxic peptide comprises the aminoacid sequence as shown in SEQIDNO:1.
Still more preferably, the aminoacid sequence of described neurotoxic peptide is for such as shown in SEQIDNO:1.
Preferably, in described step (2) or (3), described buffer is HEPES buffer solution, and described HEPES buffer solution is the 4-hydroxyethyl piperazine ethanesulfonic acid buffer solution that laboratory routine uses.
Further preferably, in described step (2) or (3), the concentration of described HEPES buffer solution is 0.02mol/L, and pH value is 7.4.
Preferably, in described step (2), the concrete operations of described supersound process are: the carbon nanomaterial first surface being in oxidation state is dissolved in HEPES buffer solution, supersound process certain hour under Ultrasound Instrument, after homogenize material dispersion, add appropriate HEPES buffer solution again, continue supersound process a period of time, then add peptide molecule.
Further preferably, the consumption of described appropriate HEPES buffer solution is that to be dissolved in final volume be in 2 ~ 3mlHEPES buffer solution to every 1mg peptide molecule.
Further preferably, the condition of described supersound process is: power is 100W, ultrasonic 15 minutes.
Preferably, in described step (3), the mode of described stirring is magnetic agitation.
Preferably, in described step (3), the molecular cut off 100kD of described ultra-filtration centrifuge tube.
Preferably, in described step (3), describedly reaction products therefrom ultra-filtration centrifuge tube is carried out step that is centrifugal, washing and comprises:
Reaction products therefrom is added ultra-filtration centrifuge tube, then centrifugal 2 ~ 4 times, each with the washing of HEPES buffer solution; Wherein, described centrifugal condition is under 5000r/m rotating speed, centrifugal 10 minutes.
Through centrifuge washing repeatedly, can by free, be not separated clean by the peptide molecule modifying carbon nanomaterial surface.
Preferably, in described step (3), the described last HEPES buffer solution having modified the carbon nanomaterial certain volume of polypeptide dissolves, and after supersound process a period of time, is then placed in 4 degree of Refrigerator stores for subsequent use.
Further preferably, in described step (3), the condition of described supersound process is: power 100W, ultrasonic 10 minutes.
Preferably, the size of carbon nanomaterial that described polypeptide is directly modified is 30 ~ 60nm.
The preparation method of the carbon nanomaterial that the polypeptide that the present invention also provides directly is modified adopts the peptide molecule of reproducibility as reducing agent, remain the biological function of peptide molecule simultaneously, eliminating in traditional chemical modification will first to the step of carbon nanomaterial and biomolecule activation processing, simplify operating procedure, save production cost.
In addition, in the process that traditional protein macromolecule is modified carbon nanomaterial, because the molecular weight of albumen is large, space structure is complicated, the impact of its surface nature on carbon nanomaterial is large, easily causes nano material aggregate and precipitate.The peptide molecule that the present invention adopts has less size, and structure is more flexible, with low cost; And peptide molecule affects less to the surface nature (as electric charge, size) etc. of modifying rear carbon nanomaterial, therefore, the stability that peptide molecule modifies rear carbon nanomaterial is better.
The third aspect, present invention also offers the carbon nanomaterial that a peptide species directly modifies and is preparing the application in tumour medicine, photoacoustic imaging contrast agent or photo-thermal therapy preparation.
The carbon nanomaterial that polypeptide provided by the invention is directly modified has multi-functional: the selectively targeted function not only with peptide molecule tumor tissues, simultaneously due to the optical property of carbon nanomaterial excellence, therefore, can be applicable to prepare tumour medicine, photoacoustic imaging contrast agent or photo-thermal therapy preparation.
The peptide species that the invention provides directly modify carbon nanomaterial, and its preparation method and application, there is following beneficial effect:
(1) peptide modified carbon nanomaterial provided by the invention not only has good biological activity, and has good stability and dispersibility;
(2) preparation method of the carbon nanomaterial that the polypeptide that the present invention also provides directly is modified adopts the peptide molecule of reproducibility as reducing agent, eliminating in traditional chemical modification will first to the step of carbon nanomaterial and biomolecule activation processing, simplify operating procedure, save production cost;
(3) present invention also offers the carbon nanomaterial that a peptide species directly modifies and prepare the application in tumour medicine, photoacoustic imaging contrast agent or photo-thermal therapy preparation.
Accompanying drawing explanation
The uv-visible absorption spectra figure of the peptide modified carbon nano-particle that Fig. 1 provides for the embodiment of the present invention 2;
The transmission electron microscope picture of the peptide modified carbon nano-particle that Fig. 2 ~ Fig. 4 provides for the embodiment of the present invention 2;
The carbon nano-particle that Fig. 5 provides for the embodiment of the present invention 1 and the peptide modified carbon nano-particle water solublity pictorial diagram that embodiment 2 provides.
Detailed description of the invention
Below in conjunction with the accompanying drawing in the embodiment of the present invention, be clearly and completely described the technical scheme in the embodiment of the present invention, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art, not making the every other embodiment obtained under creative work prerequisite, belong to the scope of protection of the invention.
Prepare carbonoxide nano-particle
Embodiment 1
Present embodiments provide a kind of preparation method of carbonoxide nano-particle, comprise the steps:
1) above the flame of wax candle, collect not clean-burning black smog with the ceramic tile that clean surface is smooth, the atrament be attached on ceramic tile (wax candle ash, main component is carbon) medication spoon is scraped and is stored in vial;
2) take out the wax candle ash of certain mass, add equal-volume DMF solvent and concentrated nitric acid, at a certain temperature back flow reaction a period of time;
3) be separated by gradient centrifugation by step (2) gained reactant liquor, the step that described gradient centrifugation is separated is:
A) slow-speed of revolution 6000 revs/min, centrifugal 5 minutes, by the removing of large-sized granule precipitation, collects upper strata black liquor and ultrasonic disperse process 2 hours;
B) with diluted sodium hydroxide solution, the pH value of step (a) gained black liquor is adjusted to neutrality, then by liquid through 9000 revs/min, centrifugal 5 minutes, further by granule precipitate and separate relatively large for size, collect upper strata pitchy transparency liquid;
C) by step (b) gained pitchy transparency liquid with 13000 revs/min, centrifugal 10 minutes, collect pitchy precipitation, and precipitate and standardize solution with the water dissolution of certain volume, ultrasonic disperse process 30 minutes, namely obtains oxidized form carbon nano-particle solution, and 4 degree of Refrigerator stores are for subsequent use.
Reduction-oxidation carbon nano-particle
Embodiment 2
Present embodiments provide a kind of preparation method adopting neurotoxic peptide (chlorotoxin, CTX) the carbonoxide nano-particle of reproducibility, comprise the steps:
1) getting the carbon nanomaterial that surface prepared by 0.5mg embodiment 1 is in oxidation state is dissolved in 1mLHEPES buffer solution, supersound process 15 minutes, material is allowed fully to disperse, then the HEPES buffer solution of 1.5mL volume is added, continue supersound process 5 minutes, then add the CTX of 4mg, obtain mixture, wherein, the aminoacid sequence of described CTX is as shown in SEQIDNO:1;
2) by step (1) gained mixture under 25 degree of room temperature, magnetic agitation reacts 24 hours, obtains product, has the carbon nanomaterial of peptide molecule in described product containing finishing;
3) by step (2) gained product under 5000r/m rotating speed, centrifugalize 10 minutes, repeatedly all rinse with HEPES buffer solution for centrifugal 3 times at every turn, what make to dissociate is not removed clean by the polypeptide modifying material surface, the HEPES buffer solution of the carbon nanomaterial 1mL volume of peptide molecule finally finishing is had to dissolve and standardize solution, then supersound process 10 minutes, obtain the carbonoxide nano-particle that described CTX directly modifies, size is 30 ~ 60nm, is placed in 4 degree of Refrigerator stores for subsequent use.
For absolutely proving beneficial effect of the present invention, present invention also offers the uv-visible absorption spectra figure of polypeptide (CTX) the modified carbon nano-particle that embodiment 2 provides, as shown in Figure 1.Curve 1 has 1 characteristic absorption peak respectively at about 230nm and 278nm place in FIG, the characteristic absorption peak of about 230nm is the characteristic peak of carbon nano-particle (CNPs) after CTX modifies, the characteristic absorption peak at 278nm place is the characteristic peak of CTX polypeptide, and the carbonoxide nano-particle that the embodiment of the present invention 2 has prepared CTX and directly modifies is described.
For further illustrating beneficial effect of the present invention, present invention also offers the transmission electron microscope picture of the peptide modified carbon nano-particle that embodiment 2 provides, as shown in figs. 2 to 4, from Fig. 2 ~ 4, illustrate that the carbonoxide nano-particle that CTX prepared by the embodiment of the present invention 2 directly modifies has good dispersibility;
For illustrating further beneficial effect of the present invention, present invention also offers carbon nano-particle that embodiment 1 provides and the peptide modified carbon nano-particle water solublity pictorial diagram that embodiment 2 provides, as shown in Figure 5, as shown in Figure 5: dispersibility and the water solublity of peptide modified front and back nano material are well kept, not significant change; Secondly, after modifying, the color of carbon nanomaterial changes carbon black color (test tube 2) (carbon black color is the unoxidized color of material with carbon element) into from dark brown (test tube 1), this also further demonstrate that CTX polypeptide can be carbon nano materials modified from the side, can reduce surface to be in the carbon nanomaterial of oxidation state.
Embodiment 3 ~ 6
The present embodiment 3 ~ 7 is substantially identical with the step of embodiment 2, and difference is only that the carbon nanomaterial of step (1) oxidation state is different from the mass ratio of CTX consumption, distinguishes as shown in the table:
Embodiment 8
Present embodiments provide a kind of preparation method adopting the Nisin modified carbonoxide nano-particle of reproducibility, comprise the steps:
1) getting the carbon nanomaterial that surface prepared by 0.5mg embodiment 1 is in oxidation state is dissolved in 1mLHEPES buffer solution, supersound process 15 minutes, material is allowed fully to disperse, then the HEPES buffer solution of 1.5mL volume is added, continue supersound process 5 minutes, then add the nisin of 4mg, obtain mixture, wherein, No. PubChemCommpoundCID of described nisin is 16219761;
2) by step (1) gained mixture under 25 degree of room temperature, magnetic agitation reacts 24 hours, obtains product;
3) by step (2) gained product under 5000r/m rotating speed, centrifugalize 10 minutes, repeatedly all rinse with HEPES buffer solution for centrifugal 3 times at every turn, what make to dissociate is not removed clean by the polypeptide modifying material surface, the HEPES buffer solution of the carbon nanomaterial 1mL volume of peptide molecule finally finishing is had to dissolve and standardize solution, then supersound process 10 minutes, obtain the carbonoxide nano-particle that described nisin is directly modified, size is 30 ~ 60nm, is placed in 4 degree of Refrigerator stores for subsequent use.
Embodiment 9
Present embodiments provide a kind of preparation method adopting c (RGDfK) cyclic peptide of reproducibility to modify carbonoxide nano-particle, comprise the steps:
1) getting the carbon nanomaterial that surface prepared by 0.5mg embodiment 1 is in oxidation state is dissolved in 1mLHEPES buffer solution, supersound process 15 minutes, material is allowed fully to disperse, then the HEPES buffer solution of 1.5mL volume is added, continue supersound process 5 minutes, then add c (RGDfK) cyclic peptide of 4mg, obtain mixture, wherein, No. PubChemCommpoundCID of described c (RGDfK) cyclic peptide is 10196873;
2) by step (1) gained mixture under 25 degree of room temperature, magnetic agitation reacts 24 hours, obtains product;
3) by step (2) gained product under 5000r/m rotating speed, centrifugalize 10 minutes, repeatedly all rinse with HEPES buffer solution for centrifugal 3 times at every turn, what make to dissociate is not removed clean by the polypeptide modifying material surface, the HEPES buffer solution of the carbon nanomaterial 1mL volume of peptide molecule finally finishing is had to dissolve and standardize solution, then supersound process 10 minutes, obtain the carbonoxide nano-particle that described c (RGDfK) cyclic peptide is directly modified, size is 30 ~ 60nm, is placed in 4 degree of Refrigerator stores for subsequent use.
The above is the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications are also considered as protection scope of the present invention.
Claims (10)
1. the carbon nanomaterial directly modified of a peptide species, the carbon nanomaterial that described polypeptide is directly modified comprises peptide molecule and carbon nanomaterial, wherein, described peptide molecule is connected by covalent bond with carbon nanomaterial, it is characterized in that, the product of carbon nanomaterial obtained by the carbon nanomaterial of employing peptide molecule reduction-oxidation state that described polypeptide is directly modified, wherein, described peptide molecule contains reproducibility aminoacid, the carbon nanomaterial of described oxidation state contains surface active groups, the surface active groups of the carbon nanomaterial of described oxidation state is carboxyl, epoxy radicals or hydroxyl.
2. the carbon nanomaterial directly modified of polypeptide as claimed in claim 1, it is characterized in that, the carbon nanomaterial of described oxidation state is carbonoxide nano-particle, graphene oxide or oxide/carbon nanometer tube.
3. the carbon nanomaterial directly modified of polypeptide as claimed in claim 1, it is characterized in that, described reproducibility aminoacid is TYR, cysteine, histidine or tryptophan.
4. the carbon nanomaterial directly modified of polypeptide as claimed in claim 1, it is characterized in that, the size of the carbon nanomaterial that described polypeptide is directly modified is 30 ~ 60nm.
5. the peptide species preparation method of carbon nanomaterial of directly modifying, is characterized in that, comprise the steps:
1) carbon nanomaterial that peptide molecule that mass ratio is 1 ~ 4:1 ~ 20 and surface are in oxidation state is got, wherein, described peptide molecule contains reproducibility aminoacid, described surface be in oxidation state containing surface active groups, the surface active groups of the carbon nanomaterial of described oxidation state is carboxyl, epoxy radicals or hydroxyl;
2) carbon nanomaterial described surface being in oxidation state adds in buffer, after supersound process, add described peptide molecule and mixing obtain mixture;
3) gained mixture stirring reaction 8 ~ 24 hours at normal temperatures, reaction products therefrom ultra-filtration centrifuge tube adds buffer and supersound process, obtains the carbon nanomaterial that described polypeptide is directly modified after carrying out centrifugal, washing.
6. the polypeptide as claimed in claim 5 preparation method of carbon nanomaterial of directly modifying, it is characterized in that, in described step (1), the carbon nanomaterial of described oxidation state is carbonoxide nano-particle, graphene oxide or oxide/carbon nanometer tube.
7. the polypeptide as claimed in claim 5 preparation method of carbon nanomaterial of directly modifying, it is characterized in that, in described step (1), described is neurotoxic peptide, RGD cyclic peptide or antibacterial polypeptide containing the amino acid whose peptide molecule of reproducibility.
8. the polypeptide as claimed in claim 5 preparation method of carbon nanomaterial of directly modifying, it is characterized in that, in described step (1), described reproducibility aminoacid is TYR, cysteine, histidine or tryptophan.
9. the polypeptide as claimed in claim 5 preparation method of carbon nanomaterial of directly modifying, it is characterized in that, in described step (3), the size of the carbon nanomaterial that described polypeptide is directly modified is 30 ~ 60nm.
10. the carbon nanomaterial that a peptide species is directly modified is preparing the application in tumour medicine, photoacoustic imaging contrast agent or photo-thermal therapy preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410201129.9A CN105079806B (en) | 2014-05-13 | 2014-05-13 | A kind of carbon nanomaterial and its preparation method and application that polypeptide is directly modified |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410201129.9A CN105079806B (en) | 2014-05-13 | 2014-05-13 | A kind of carbon nanomaterial and its preparation method and application that polypeptide is directly modified |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105079806A true CN105079806A (en) | 2015-11-25 |
| CN105079806B CN105079806B (en) | 2018-12-28 |
Family
ID=54561720
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410201129.9A Active CN105079806B (en) | 2014-05-13 | 2014-05-13 | A kind of carbon nanomaterial and its preparation method and application that polypeptide is directly modified |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105079806B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106349331A (en) * | 2016-08-23 | 2017-01-25 | 国家纳米科学中心 | Bispyrene-based pH (Potential of Hydrogen)-response self-assembly polypeptide nano material, and preparation method and application thereof |
| CN108126207A (en) * | 2018-03-15 | 2018-06-08 | 广西中医药大学 | graphene oxide target medicine carrier material and its preparation method and application |
| CN109745578A (en) * | 2018-08-31 | 2019-05-14 | 京东方科技集团股份有限公司 | Bone renovating material and preparation method thereof |
| CN113440607A (en) * | 2021-06-30 | 2021-09-28 | 澳门大学 | Full-target antigen presenting cell tumor vaccine and preparation method and application thereof |
| CN113797356A (en) * | 2020-06-17 | 2021-12-17 | 广东量子墨滴生物科技有限公司 | Carbon nano composite biological preparation, preparation method and application |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007016466A2 (en) * | 2005-07-29 | 2007-02-08 | Sloan-Kettering Institute For Cancer Research | Single wall nanotube constructs and uses therefor |
| CN1948146A (en) * | 2006-11-09 | 2007-04-18 | 上海交通大学 | Method of modifying multiwall carbon nano-tube |
| CN101670108A (en) * | 2009-08-13 | 2010-03-17 | 苏州纳米技术与纳米仿生研究所 | Medicine carrying system based on nano graphene oxide |
| US20100074845A1 (en) * | 2008-09-02 | 2010-03-25 | Gambhir Sanjiv S | Enhanced sensitivity carbon nanotubes as targeted photoacoustic molecular imaging agents |
| CN102274510A (en) * | 2011-07-15 | 2011-12-14 | 华南理工大学 | Preparation method of carbon nanotube-chitosan-phycocyanin nanoparticles |
| CN102370980A (en) * | 2010-08-13 | 2012-03-14 | 同济大学 | Preparation method of nanometer graphene oxide carrier for photodynamic therapy |
-
2014
- 2014-05-13 CN CN201410201129.9A patent/CN105079806B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007016466A2 (en) * | 2005-07-29 | 2007-02-08 | Sloan-Kettering Institute For Cancer Research | Single wall nanotube constructs and uses therefor |
| CN1948146A (en) * | 2006-11-09 | 2007-04-18 | 上海交通大学 | Method of modifying multiwall carbon nano-tube |
| US20100074845A1 (en) * | 2008-09-02 | 2010-03-25 | Gambhir Sanjiv S | Enhanced sensitivity carbon nanotubes as targeted photoacoustic molecular imaging agents |
| CN101670108A (en) * | 2009-08-13 | 2010-03-17 | 苏州纳米技术与纳米仿生研究所 | Medicine carrying system based on nano graphene oxide |
| CN102370980A (en) * | 2010-08-13 | 2012-03-14 | 同济大学 | Preparation method of nanometer graphene oxide carrier for photodynamic therapy |
| CN102274510A (en) * | 2011-07-15 | 2011-12-14 | 华南理工大学 | Preparation method of carbon nanotube-chitosan-phycocyanin nanoparticles |
Non-Patent Citations (3)
| Title |
|---|
| DUNCAN C. HONE，ET AL: "Generation of Cytotoxic Singlet Oxygen via Phthalocyanine-Stabilized Gold Nanoparticles: A Potential Delivery Vehicle for Photodynamic Therapy", 《LANGMUIR》 * |
| 董丽红: "光热治疗", 《两亲分子在纳米技术领域中的应用》 * |
| 韦进全 等: "《碳纳米管宏观体》", 31 May 2006, 清华大学出版社 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106349331A (en) * | 2016-08-23 | 2017-01-25 | 国家纳米科学中心 | Bispyrene-based pH (Potential of Hydrogen)-response self-assembly polypeptide nano material, and preparation method and application thereof |
| CN106349331B (en) * | 2016-08-23 | 2020-05-15 | 国家纳米科学中心 | Bipyrene-based pH response self-assembly polypeptide nano material and preparation method and application thereof |
| CN108126207A (en) * | 2018-03-15 | 2018-06-08 | 广西中医药大学 | graphene oxide target medicine carrier material and its preparation method and application |
| CN109745578A (en) * | 2018-08-31 | 2019-05-14 | 京东方科技集团股份有限公司 | Bone renovating material and preparation method thereof |
| US11517610B2 (en) | 2018-08-31 | 2022-12-06 | Beijing Boe Technology Development Co., Ltd. | Bone repair material and preparation method thereof, and biological composite scaffold |
| CN113797356A (en) * | 2020-06-17 | 2021-12-17 | 广东量子墨滴生物科技有限公司 | Carbon nano composite biological preparation, preparation method and application |
| WO2021253470A1 (en) * | 2020-06-17 | 2021-12-23 | 广东量子墨滴生物科技有限公司 | Carbon nanocomposite biological agent and preparation method and application therefor |
| CN113440607A (en) * | 2021-06-30 | 2021-09-28 | 澳门大学 | Full-target antigen presenting cell tumor vaccine and preparation method and application thereof |
| WO2023274299A1 (en) * | 2021-06-30 | 2023-01-05 | 澳门大学 | Full-target antigen-presenting cell tumor vaccine, and preparation method therefor and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105079806B (en) | 2018-12-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105079806A (en) | Carbon nanomaterial directly modified by polypeptide as well as preparation method and application thereof | |
| Tao et al. | A new type of polymer carbon dots with high quantum yield: From synthesis to investigation on fluorescence mechanism | |
| Heister et al. | Higher dispersion efficacy of functionalized carbon nanotubes in chemical and biological environments | |
| Karajanagi et al. | Protein-assisted solubilization of single-walled carbon nanotubes | |
| Zhan et al. | Spatiotemporal control of supramolecular self-assembly and function | |
| Halas | Nanoscience under glass: the versatile chemistry of silica nanostructures | |
| CN102671217B (en) | Preparation of CT/MR bimodal imaging nano contrast medium with folate targeting function | |
| Li et al. | Ionic liquid-functionalized fluorescent carbon nanodots and their applications in electrocatalysis, biosensing, and cell imaging | |
| CN104387446B (en) | A kind of graphene dispersion agent and the preparation method of graphene dispersing solution | |
| Zhou et al. | Disulfide bond: dramatically enhanced assembly capability and structural stability of tobacco mosaic virus nanorods | |
| Jiang et al. | AIE-active metal-organic coordination complexes based on tetraphenylethylene unit and their applications | |
| CN103693633A (en) | Method for green synthesis of fluorescent chiral carbon dots | |
| Tao et al. | Multiporous supramolecular microspheres for artificial photosynthesis | |
| Wang et al. | Designing a sustainable fluorescent targeting probe for superselective nucleus imaging | |
| CN103771390A (en) | Method for synthesizing carbon quantum dot by using biologic active enzyme auxiliary microwave method, carbon quantum dot prepared by using method, and application thereof | |
| CN105749876B (en) | A kind of preparation method of chitosan-modified graphene oxide quantum dot sorbing material | |
| Li et al. | Hydrogelation and crystallization of sodium deoxycholate controlled by organic acids | |
| Zhai et al. | Incorporation and recovery of SWNTs through phase behavior and aggregates transition induced by changes in pH in a catanionic surfactants system | |
| CN107037099B (en) | The preparation method of reductive amination graphene oxide modified electrode | |
| CN102818826A (en) | Electrochemical cell-based biosensor based on nanometer Ag@BSA biomimetic interface and preparation method thereof | |
| CN102989013A (en) | Preparation method for lymphatic vessel recognized carbon nanosphere | |
| Guo et al. | Effects of polymers on the properties of hydrogels constructed using sodium deoxycholate and amino acid | |
| Das et al. | Carbon quantum dots in bioimaging and biomedicines | |
| CN107055608A (en) | A kind of transparent dispersion liquid of nano zirconium dioxide and preparation method and application | |
| CN104826126A (en) | Method for preparing high mechanical strength three-component fluorescence hydrogel |
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 |