CN102657872A - Multifunctional graphene oxide/ polyamide-amine (PAMAM)/ diethylene triamine pentaacetic-gadolinium (DTPA-Gd)/ prostate stem cell antigen (PSCA) antibody material and preparation method and application thereof - Google Patents
Multifunctional graphene oxide/ polyamide-amine (PAMAM)/ diethylene triamine pentaacetic-gadolinium (DTPA-Gd)/ prostate stem cell antigen (PSCA) antibody material and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a multifunctional graphene oxide/ polyamide-amine (PAMAM)/ diethylene triamine pentaacetic-gadolinium (DTPA-Gd)/ prostate stem cell antigen (PSCA) antibody material. PAMAM dendrimers are covalently modified on nano graphene oxide flakes, and DTPA-Gd and PSCA antibodies are modified by using amino on the PAMAM dendrimers. The obtained material has very high biocompatibility and stability, can be used as a good T1 contrast medium, and has a good targeting property for prostatic cancer cells; and doxorubicin hydrochloride can be adsorbed to the graphene oxide/ PAMAM/ DTPA-Gd/ PSCA antibody material under coaction of phi-phi bonds, and the material is high in medicament loading rate and has an obvious targeting effect. The multifunctional graphene oxide/ PAMAM/ DTPA-Gd/ PSCA antibody material has targeting medicament loading and magnetic resonance imaging (MRI) functions, and has a very wide biological application prospect.
Description
Technical field
The present invention relates to a kind of graphene oxide/PAMAM/DTPA-Gd/ doxorubicin hydrochloride/psca antibody multifunctional material and preparation method thereof, belong to nano-medicament carrier and nano-contrast agent technical field.
Background technology
Carbon is the most magical element of nature; In the nanometer world; The performance of carbon is amazing too; The strong K sea nurse of the peace moral of Univ Manchester UK in 2004 (Andre K.Geim) produces Graphene first, and it is the thinnest in the world novel two-dimensional nano material, and its thickness is merely 0.35nm.Its peculiar property has attracted countless scientists, we can say that it is potentialization in many new materials, is called as " miracle material ", all has great application prospect in industry, power industry and electronic industry.
Compare with graphite; Contain chemical groups such as carboxyl, hydroxyl, carbonyl and epoxy radicals in the graphene oxide, wherein carboxyl and hydroxyl modified are in the edge of oxidized graphite flake, and carbonyl and epoxy radicals are between the graphene oxide layer; Graphite is water-fast material originally; Oxidation rear oxidation Graphene has extremely strong hydrophilic, is scattered in easily to form stable sols in the polarizable medium, and this is because the existence of these active groups of graphene oxide surface.
The 1950's, scientist has proposed the structure of graphene oxide in succession, mainly is three big basic models, Ruess model, Hofmann model and Scholz-Boehm model.It is generally acknowledged that graphene oxide is a two-dimensional layered structure, interlayer contains a large amount of hydroxyls and the acid active group of carboxyl.At present, the research of graphene oxide begins to pay close attention to its application aspect biological medicine, and such as medicine, gene transportation, particularly the research of treatment of cancer targeted drug transmission system has caused very large concern.
Summary of the invention
The purpose of this invention is to provide a kind of graphene oxide/PAMAM/DTPA-Gd/ doxorubicin hydrochloride/psca antibody multifunctional material.This pharmaceutical carrier good dispersion in water, good biocompatibility, the toxicity of material own is little, and the drug loading ability is strong, in nuclear magnetic resonance, has good application.
Another object of the present invention is for this difunctional preparation of drug carriers method is provided, and that this method has is simple to operate, raw material is easy to get and advantage such as with low cost.
The objective of the invention is to realize like this:
The method for preparing of graphene oxide/PAMAM/DTPA-Gd/ doxorubicin hydrochloride/psca antibody multifunctional material comprises the steps:
(1) connect PAMAM on the nano graphene oxide surface: nano graphene oxide is dispersed in the alcohol and activation; With containing the third generation is that the alcoholic solution of the polyamide-amide dendritic macromole of kernel mixes and stirs 18～30hr with the trimesic acid, obtains the GO/PAMAM nano material; Nano graphene oxide and third generation polyamide-amide dendritic macromole weight ratio are 1:20～40;
(2) the GO/PAMAM nano material is dissolved in the organic solvent; Add triethylamine and diethylene-triamine pentaacetic acid dianhydride, 70～85 ℃ are heated 20～60min down, stop to heat continued reaction 10～18hr; Get washing of precipitate, obtain the GO/PAMAM/DTPA nano material; The weight ratio of GO/PAMAM nano material and diethylene-triamine pentaacetic acid dianhydride is 4～8:1;
(3) the GO/PAMAM/DTPA nano material is with after solubility gadolinium salt mixes, and 35～50 ℃ of insulation 10～15hr under the condition of secluding air get deposition water dialysis 12～30hr, obtain graphene oxide/PAMAM/DTPA-Gd nano material; Gadolinium element and GO/PAMAM/DTPA nano material amount ratio are 0.002～0.01mmol/mg;
(4) connect anti-psca antibody: graphene oxide/PAMAM/DTPA-Gd nano material is reacted 10～16hr and washing with anti-carcinoma of prostate stem cell antigen antibody PSCA down at 0～6 ℃ in the phosphate buffer of pH=7.2～7.5 after glutaraldehyde is handled; Graphene oxide/PAMAM-DTPA-Gd nano material and anti-carcinoma of prostate stem cell antigen antibody weight ratio are 10000:1～10000:5.
Nano graphene oxide thickness in the step (1) is 1～2nm, and the surface has carboxyl.
The nano graphene oxide method for preparing comprises the steps: that adding 95wt%～98wt% concentrated sulphuric acid after graphene powder expands with saline solution stirs 6～12hr, adds KMnO again
4, mixture stirs 25～40min down at 35～40 ℃, stirs 40～60min down at 65～80 ℃, stirs 20～35min down at 98 ~ 105 ℃ then; Use the hydrogen peroxide cessation reaction, washing is got solid, ultra-sonic dispersion in water.
In the step (1); With 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide DEC and N-hydroxy succinic acid imines NHS activation nano graphene oxide, nano graphene oxide, 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide and N-hydroxy succinic acid imines weight ratio are 1:3～8:3～8; Alcohol described in the step (1) is methanol.
In the step (2), said organic solvent is dimethyl formamide DMF; The weight ratio of GO/PAMAM nano material and triethylamine is 1:4～8.
Can obtain graphene oxide/PAMAM/DTPA-Gd/PSCA antibody multifunctional material through said method.
This graphene oxide/PAMAM/DTPA-Gd/PSCA antibody multifunctional material is used for the treatment cancer targeting and the nuclear magnetic resonance, NMR T of may command drug release
1Contrast agent.
Further can prepare graphene oxide/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride composite, step comprises:
With graphene oxide/PAMAM/DTPA-Gd/PSCA antibody multifunctional material with contain doxorubicin hydrochloride, pH=7.2～7.5 phosphate buffers stirring reaction 16～30hr under the lucifuge condition; Doxorubicin hydrochloride and graphene oxide/PAMAM/DTPA-Gd/PSCA antibody multifunctional material weight ratio is 1:1～1:2, is preferably 2:3.
The present invention at first adopts improved Hummers legal system to be equipped with graphene oxide; Then with the dendritic macromole (PAMAM) of this graphene oxide finishing 3G, graphene oxide and be to be connected through generating the amido link covalency between the dendritic macromole (PAMAM) of kernel with the trimesic acid; Utilize amido modified DTPA-Gd and psca antibody above the dendroid, doxorubicin hydrochloride interacts through pi-pi bond and is adsorbed onto on the graphene oxide.Graphene oxide has bilateral structure, and there are a lot of active groups on the surface, and extraordinary biocompatibility and stability are arranged, and the drug loading ability is strong, can be good at carrying out drug loading and transportation, can be used as good T in the modification behind the DTPA-Gd
1Contrast agent, thus realize synthetic purpose with multifunctional material of targeted therapy and MRI imaging.The gained material has excellent biological compatibility simultaneously, helps the application of material at biomedicine field.
Essential implementation of the present invention is: when adopting Hummers method and supercritical ultrasonics technology to prepare graphene oxide; Note adding the amount of potassium permanganate and the grasp of time; To add potassium permanganate with the time about 1 hour; The last ultransonic time will be grown, and the graphene oxide lamella of preparation is thinner like this, and evenly.Temperature is crucial when meeting DTPA-Gd, guarantee that the temperature in each step is accurate, and heating and cooling rapidly are all unfavorable to the preparation of material.The material that our this method is prepared can be scattered in the water well, and has excellent biological compatibility.To equipment require low; The needed raw material price is low; By-product is nuisanceless, environmentally safe.The present invention is creative, novelty and extensive practicality is being arranged aspect cancer and the oncotherapy.
Advantage of the present invention is:
1. the graphene oxide that makes/PAMAM/DTPA-Gd/ doxorubicin hydrochloride/psca antibody multifunctional material, the drug loading amount is big, and the targeting effect is obvious, and the kill cancer cell effect is remarkable.
2. preparation facilities is simple;
3. thin, the good dispersion of the graphene oxide lamella of preparation, good water solubility;
Raw material be easy to get, cheap;
5. simple to operate; Environmentally safe.
6. be a kind of multifunctional material with carcinoma of prostate tumor cell specific target tropism, drug loading and transmission and NMR-imaging, big in field using values such as medicine and biologies.
Description of drawings
Fig. 1 is the infrared spectrogram (IR figure) of the graphene oxide of preparation in the embodiment of the invention 1; A among Fig. 1 is a graphite, and b is a graphene oxide;
Fig. 2 is the AFM picture (AFM) of the graphene oxide of preparation in the embodiment of the invention 1;
Fig. 3 is the transmission electron microscope photo (TEM) of the graphene oxide of preparation in the embodiment of the invention 1;
Fig. 4 is the graphene oxide/PAMAM (a) of preparation in the embodiment of the invention 2 and the amino density analysis of graphene oxide/PAMAM/DTPA material (b);
Fig. 5 is weighted to the linear relationship chart (b) between the gadolinium concentration in image pattern (a) and relaxation rate R1 (1/T1) and the sample for the graphene oxide/T1 of PAMAM/DTPA-Gd/PSCA dispersion of materials in water in the embodiment of the invention 3;
Fig. 6 is weighted to the linear relationship chart (b) between the gadolinium concentration in image pattern (a) and relaxation rate R2 (1/T2) and the sample for the graphene oxide/T2 of PAMAM/DTPA-Gd/PSCA dispersion of materials in water in the embodiment of the invention 3;
Fig. 7 is the MTT toxicity data of graphene oxide in the embodiment of the invention 3/PAMAM/DTPA-Gd/PSCA material and PC-3 (a) and two kinds of cells of MCF-7 (b);
Fig. 8 is weighted to image pattern (a), signal strength map (b) and the ionic amount of cytophagy Gd (c) for the T1 of graphene oxide/PAMAM/DTPA-Gd/PSCA antibody material in the embodiment of the invention 3 and PC-3 and two kinds of cells of MCF-7;
Fig. 9 is respectively the ultraviolet-visible light spectrogram (a) and the average drug loading scattergram (b) of graphene oxide in the embodiment of the invention 4/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride material drug carrier amount.Can know that through calculating average drug loading is 0.7415 milligram/milligram;
Figure 10 is graphene oxide in the embodiment of the invention 4/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride material medicine cumulative release figure when pH=7.4 and pH=5.3;
Figure 11 is respectively the MTT toxicity data of embodiment 4 graphene oxides/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride and doxorubicin hydrochloride and PC-3 and two kinds of cells of MCF-7; (a) and (b) be respectively the MTT that graphene oxide/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride and doxorubicin hydrochloride and PC-3 cell are hatched, (c) be respectively the MTT that graphene oxide/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride and doxorubicin hydrochloride and MCF-7 cell are hatched (d).
The specific embodiment
In order to understand essence of the present invention better, specify technology contents of the present invention through embodiment below, but content of the present invention is not limited thereto.
1) takes by weighing the 1g graphene powder in beaker, add 50g sodium chloride again, stir 10min; In beaker, add an amount of distilled water, carry out sucking filtration and collect expansible graphene powder;
2) the expansible graphene powder that will collect places single neck flask of 250mL, adds 98% the concentrated sulphuric acid of 23mL, and magnetic agitation 8h slowly progressively adds 3g KMnO
4, keep temperature therebetween less than 20 ℃;
3) mixture stirs 30min at 35～40 ℃ of lower magnetic forces, stirs 45min at 65～80 ℃ of lower magnetic forces then.After reacting by heating finishes, add the distilled water of 46mL in single neck flask, mixture is at 98～105 ℃ of lower magnetic forces stirring 30min; 30% the H that adds distilled water and the 10mL of 140mL
2O
2, cessation reaction;
4) with 5% HCl and distilled water with mixture centrifugal repeatedly and filtering and washing, product is dispersed in ultrasonic 1h in the water, obtain nano graphene oxide, solution colour is brown.
Fig. 1 is the infrared spectrogram (IR figure) of the graphene oxide of preparation among the embodiment 1, from infrared spectrogram, sees, at 1723cm
-1C=O stretching vibration on the carboxyl appears in the place, explains that graphite powder is oxidized to graphene oxide, and the surface has carboxyl; A among Fig. 1 is a graphite, and b is a graphene oxide.
Fig. 2 is the AFM picture (AFM) of the graphene oxide of preparation among the embodiment 1; From AFM figure, can find out very clearly that the graphene oxide sheet is level and smooth; And advanced Measurement and analysis and can know that the thickness of graphene platelet approximately was 1.246nm; The thickness of the single-layer graphene of reporting with document coincide, and shows the graphene oxide thin slice successfully preparation of monolayer;
Fig. 3 is the transmission electron microscope photo (TEM) of the graphene oxide of preparation among the embodiment 1; On scheming, can find out that the structure of graphene platelet shape is very obvious; And on the whole Graphene is not smooth fully; But lamella is very thin, locates to exist comparatively significantly fold and the generation of folding phenomenon on the edge of, and this is because the thermodynamics fluctuation of two dimensional crystal is caused.
1) takes by weighing graphene oxide GO 40mg in beaker, add the 100mL absolute methanol, add 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide EDC and each 0.2g of N-hydroxy succinic acid imines NHS, ultrasonic activation 1h;
Take by weighing PAMAM (3G) 1g in addition in the 50mL beaker, add the 30ml absolute methanol, ultra-sonic dispersion 1h;
Ultra-sonic dispersion slowly is added drop-wise to PAMAM (3G) solution in the GO solution after finishing, and stirs 24h under the room temperature;
Reaction is centrifugal after finishing, and (12000rpm 12min) washs, and makes GO/PAMAM (3G) nano material;
2) take by weighing 20mg GO/PAMAM nano material, be dissolved in the 8mL dry DMF;
Add 0.18mL triethylamine (130mg), add 4mg diethylene-triamine pentaacetic acid dianhydride (DTPA) again, place the 25mL flask, slowly be heated to 80 ℃ then, continue 30min, remove thermal source, one night of stirring at normal temperature;
Reacted material is separated with centrifugal method; Earlier with the DMF washing that contains TEA 1% 3 times; Wash 3 times, acetone is washed 3 times again, is stored in after the drying in the cryogenic environment; Obtain connecting the GO/PAMAM nano material of divinyl five amine acetic acid parts (DTPA), i.e. GO/PAMAM (3G)-DTPA nano material;
3) 20mg GO/PAMAM-DTPA nano material and 10ml pH=8 buffer are mixed in the 25mL flask, add 40mg Gd (NO again
3)
3.6H
2O (0.09mmol) fully mixes, and again with the reactant liquor good seal, is heated to 40 ℃ lentamente, one night of constant temperature;
Reactant liquor is centrifugal, with the precipitate 24h that in distilled water, dialyses, remove responseless gadolinium ion, promptly get the GO/PAMAM-DTPA-Gd nano material.
Fig. 4 is the graphene oxide/PAMAM (a) of preparation among the embodiment 2 and the amino density analysis of graphene oxide/PAMAM/DTPA material (b), records the amino density of GO/PAMAM nano-material surface and is about 1.152 * 10
-4Mol/g, the surface amino groups density that records the GO/PAMAM-DTPA nano material is 6.17 * 10
-5Mol/g, experimental result can be found out thus, nano-material surface is amino to also have residue, can be used for connecing targeted molecular.
1) weighs 5mg embodiment 2 resulting graphene oxides/PAMAM-DTPA-Gd nano material dispersion of materials in the PBS that contains 5% glutaraldehyde (pH=7.4) buffer solution; Leave standstill 2h; Come out the material centrifugalize,, be dispersed in again then in PBS (pH=7.4) buffer solution with PBS washing three times; Add 10 μ L (concentration 0.13 μ g/ μ L) psca antibody, graphene oxide/PAMAM-DTPA-Gd nano material is cultivated with antibody;
2) stir 12h at 4 ℃;
3) wash for several times with PBS (pH=7.4) buffer solution after reaction finishes, be stored in 4 ℃ PBS (pH=7.4) buffer solution.The graphene oxide that obtains/PAMAM/DTPA-Gd/PSCA antibody multifunctional material has good water-solubility.
Fig. 5 is weighted to the linear relationship chart (b) between the gadolinium concentration in image pattern (a) and relaxation rate R1 (1/T1) and the sample for the T1 that graphene oxide/PAMAM/DTPA-Gd/PSCA antibody multifunctional material among the embodiment 3 is dispersed in the water;
Fig. 6 is weighted to the linear relationship chart (b) between the gadolinium concentration in image pattern (a) and relaxation rate R2 (1/T2) and the sample, T for the T2 that graphene oxide/PAMAM/DTPA-Gd/PSCA antibody multifunctional material among the embodiment 3 is dispersed in the water
1And T
2Imaging signal intensity changes along with the increase of Gd ion concentration, is embodied in: T
1Imaging signal intensity is along with the increase image of Gd ion concentration obviously brightens; T
2Imaging signal intensity is along with the obvious deepening of increase image of Gd ion concentration.Relaxation rate is along with the increase of sample concentration is linear growth, and the longitudinal relaxation coefficient is r
1Be 8.34Gd Mm
-1s
-1, transverse relaxation coefficient r
2Be 12.47Gd Mm
-1s
-1, and the r under 0.5T
2/ r
1Value is 1.5, is fit to be T
1Contrast agent.These data explain that also the GO/PAMAM/DTPA-Gd/PSCA nano material can be used as a good T
1Contrast agent;
Fig. 7 is the MTT toxicity data of graphene oxide among the embodiment 3/PAMAM/DTPA-Gd/PSCA antibody multifunctional material and PC-3 and two kinds of cells of MCF-7; Along with the increase of the concentration of GO/PAMAM/DTPA-Gd/PSCA antibody multifunctional material, the vigor of PC-3 and MCF-7 cell does not take place significantly to reduce.After concentration was hatched 24 hours up to the sample of 200 μ g/mL and PC-3 and two kinds of cells of MCF-7, two kinds of cells still kept the cell viability greater than 85%.These results show that GO/PAMAM/DTPA-Gd/PSCA antibody multifunctional material cytotoxicity to PC-3 and two kinds of cells of MCF-7 under such concentration is lower.
Fig. 8 is weighted to image pattern (a) for the graphene oxide/PC-3 of PAMAM/DTPA-Gd/PSCA antibody nano material labelling among the embodiment 3 and the T1 of two kinds of cells of MCF-7; Signal strength map (b) and the ionic amount of cytophagy Gd (c); Along with the T1 weighted imaging of two kinds of cells of increase of Gd ion concentration brightens gradually; Signal intensity increases gradually; Analysis-by-synthesis finds out that the T1 weighted imaging reinforced effects of high expressing cell PC-3 is more obvious, and this explains the good targeting of psca antibody on the one hand, and illustrative material is well suited for the contrast agent as T1 on the one hand.
Embodiment 4
1) the GO/PAMAM-DTPA-Gd/PSCA antibody multifunctional material of getting 30mg embodiment 3 preparation in the PBS of the doxorubicin hydrochloride of 20mL solution under (pH=7.4) room temperature lucifuge stir 24h;
2) centrifugalize, using the centrifugal resulting solid of PBS solution washing is colourless to centrifugal liquid, vacuum drying obtains GO/PAMAM-DTPA-Gd/PSCA antibody/doxorubicin hydrochloride medicine transmission material under the room temperature;
3) remaining medicament contg is through measuring the uv absorption of centrifugal liquid (containing cleaning mixture), and the absorption value that is taken at the 490nm place is calculated.Thereby also can calculate the amount that is loaded in the medicine on graphene oxide/PAMAM-DTPA-Gd/PSCA material.Parallel three experiments, drug loading is averaged.
The calculating of doxorubicin hydrochloride load capacity is through measuring the uv absorption of centrifugal liquid (containing cleaning mixture) and original solution, and the absorption value that is taken at the 490nm place is calculated.
Fig. 9 is respectively the ultraviolet figure and average drug loading scattergram of graphene oxide among the embodiment 4/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride material drug carrier amount.The difference that absorbs according to the UV-vis before and after medicine and the graphene oxide/PAMAM/DTPA-Gd effect can calculate drug loading.Can know that through calculating average drug loading is 0.74 milligram/milligram;
Figure 10 is graphene oxide among the embodiment 4/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride material medicine cumulative release 100% figure when pH=7.4 and pH=5.3; As can be seen from the figure; The release of the DOX of medicament-carried GO/PAMAM/DTPA-Gd/PSCA hybrid material is the pH value response and lasting, can accomplish that controllability discharges the effect that gentle slow release is put medicine.When pH=7.4, the burst size of DOX is slow, and seldom; Medicine has only discharged 6.5% during 24h, during 72h drug release 8%, yet when pH=5.3; The burst size of DOX increases, and medicine has discharged 38.5% during 24h, behind 72h; The release of medicine has reached 40%, and this is that hydrophilic and the dissolubility of DOX increase greatly because under acid condition;
Figure 11 is respectively the MTT toxicity data after graphene oxide/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride material and doxorubicin hydrochloride and PC-3 and two kinds of cells of MCF-7 are hatched altogether; (a) and (b) be respectively the MTT data that graphene oxide/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride and doxorubicin hydrochloride and PC-3 cell are hatched, (c) be respectively the MTT data that graphene oxide/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride and doxorubicin hydrochloride and MCF-7 cell are hatched (d).
Can see from the result; Graphene oxide/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride multifunctional material is better than simple doxorubicin hydrochloride to the fragmentation effect of cancerous cell; And; High expressed and low cancerous cell of expressing all there are fragmentation effect, but obvious especially to the cancerous cell fragmentation effect of high expressed.
Claims (9)
1. the method for preparing of graphene oxide/PAMAM/DTPA-Gd/PSCA antibody multifunctional material is characterized in that, comprises the steps:
(1) connect PAMAM on the nano graphene oxide surface: nano graphene oxide is dispersed in the alcohol and activation; With containing the third generation is that the alcoholic solution of the polyamide-amide dendritic macromole of kernel mixes and stirs 18～30hr with the trimesic acid, obtains the GO/PAMAM nano material; Nano graphene oxide and third generation polyamide-amide dendritic macromole weight ratio are 1:20～40;
(2) the GO/PAMAM nano material is dissolved in the organic solvent; Add triethylamine and diethylene-triamine pentaacetic acid dianhydride, 70～85 ℃ are heated 20～60min down, stop to heat continued reaction 10～18hr; Get washing of precipitate, obtain the GO/PAMAM/DTPA nano material; The weight ratio of GO/PAMAM nano material and diethylene-triamine pentaacetic acid dianhydride is 4:1～8:1;
(3) the GO/PAMAM/DTPA nano material is with after solubility gadolinium salt mixes, and 35～50 ℃ of insulation 10～15hr under the condition of secluding air get deposition water dialysis 12～30hr, obtain graphene oxide/PAMAM/DTPA-Gd nano material; Gadolinium element and GO/PAMAM/DTPA nano material amount ratio are 0.002～0.01mmol/mg;
(4) connect anti-psca antibody: the GO/PAMAM/DTPA-Gd nano material is reacted 10～16hr and washing with anti-carcinoma of prostate stem cell antigen antibody down at 0～6 ℃ in the phosphate buffer of pH=7.2～7.5 after glutaraldehyde is handled; The weight ratio of using of graphene oxide/PAMAM-DTPA-Gd nano material and anti-carcinoma of prostate stem cell antigen antibody is 10000:1～10000:5.
2. the method for preparing of the said graphene oxide of claim 1/PAMAM/DTPA-Gd/PSCA antibody multifunctional material is characterized in that, the nano graphene oxide thickness in the step (1) is 1～2nm, and the surface has carboxyl;
Method for preparing comprises the steps: that adding 95wt%～98wt% concentrated sulphuric acid after graphene powder expands with saline solution stirs 6～12hr, adds KMnO again
4, mixture stirs 25～40min down at 35～40 ℃, stirs 40～60min down at 65～80 ℃, stirs 20～35min down at 98～105 ℃ then; Use the hydrogen peroxide cessation reaction, washing is got solid, ultra-sonic dispersion in water.
3. the method for preparing of the said graphene oxide of claim 1/PAMAM/DTPA-Gd/PSCA antibody multifunctional material; It is characterized in that; With 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide and N-hydroxy succinic acid imines activation nano graphene oxide, nano graphene oxide, 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide and N-hydroxy succinic acid imines weight ratio are 1:3～8:3～8 in the step (1); Alcohol described in the step (1) is methanol.
4. the method for preparing of the said graphene oxide of claim 1/PAMAM/DTPA-Gd/PSCA antibody multifunctional material is characterized in that, organic solvent is a dimethyl formamide described in the step (2); The weight ratio of GO/PAMAM nano material and triethylamine is 1:4～8.
5. graphene oxide/PAMAM/DTPA-Gd/PSCA antibody multifunctional material is characterized in that, through each described method preparation of claim 1～4.
6. graphene oxide/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride multifunctional material; It is characterized in that method for preparing comprises the steps: the said graphene oxide of claim 5/PAMAM/DTPA-Gd/PSCA antibody multifunctional material and contains doxorubicin hydrochloride, pH=7.2～7.5 phosphate buffers stirring reaction 16～30hr under the lucifuge condition; Doxorubicin hydrochloride and graphene oxide/PAMAM/DTPA-Gd/PSCA antibody multifunctional material weight ratio is 1:1～1:2.
7. the application of the said graphene oxide of claim 5/PAMAM/DTPA-Gd/PSCA antibody multifunctional material aspect the preparation NMR contrast agent.
8. the said graphene oxide of claim 5/PAMAM/DTPA-Gd/PSCA antibody multifunctional material is used for preparation treatment carcinoma of prostate pharmaceutical carrier.
9. the application of the said graphene oxide of claim 6/PAMAM/DTPA-Gd/PSCA antibody/doxorubicin hydrochloride multifunctional material aspect preparation treatment carcinoma of prostate medicine.
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| CN102872458A (en) * | 2012-10-24 | 2013-01-16 | 佛山市第一人民医院 | Therapeutic vaccine adjuvant |
| CN104436221A (en) * | 2013-09-23 | 2015-03-25 | 中国科学院苏州纳米技术与纳米仿生研究所 | Grapheme-oxide based contrast agent and preparation method thereof |
| CN105153373A (en) * | 2015-09-21 | 2015-12-16 | 扬州市祥华新材料科技有限公司 | Preparation method for modified poly(ammonia ester-acrylic ester) emulsion of polyfunctionality compound and application |
| CN105457681A (en) * | 2015-10-26 | 2016-04-06 | 常州大学 | Preparation method of ZnO/graphene oxide composite photocatalysis material |
| GB2532449A (en) * | 2014-11-19 | 2016-05-25 | Univ Manchester | Use of Nanomaterials in treating cancer |
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| CN110665486A (en) * | 2019-10-17 | 2020-01-10 | 福州大学 | Magnetic ferroferric oxide-PAMAM-antibody complex and preparation method and application thereof |
| CN110856749A (en) * | 2018-08-23 | 2020-03-03 | 胡尚秀 | Boron-doped graphene quantum dot-containing nanocomposite and application thereof |
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| CN102872458A (en) * | 2012-10-24 | 2013-01-16 | 佛山市第一人民医院 | Therapeutic vaccine adjuvant |
| CN104436221B (en) * | 2013-09-23 | 2018-03-20 | 中国科学院苏州纳米技术与纳米仿生研究所 | Contrast agent based on graphene oxide composite material and preparation method thereof |
| CN104436221A (en) * | 2013-09-23 | 2015-03-25 | 中国科学院苏州纳米技术与纳米仿生研究所 | Grapheme-oxide based contrast agent and preparation method thereof |
| WO2015039555A1 (en) * | 2013-09-23 | 2015-03-26 | 中国科学院苏州纳米技术与纳米仿生研究所 | Contrast agent based on graphene oxide material and preparation method therefor |
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| CN106496585A (en) * | 2016-10-21 | 2017-03-15 | 浙江大学 | Sensitive graphene oxide based nano-materials of a kind of ROS and its preparation method and application |
| CN106496585B (en) * | 2016-10-21 | 2019-02-19 | 浙江大学 | A kind of graphene oxide based nano-material and its preparation method and application of ROS sensitivity |
| CN106801040A (en) * | 2017-02-02 | 2017-06-06 | 复旦大学 | Hydrophilic graphene film material of antibody modification and preparation method thereof and the application in cell capture |
| CN107375241A (en) * | 2017-08-01 | 2017-11-24 | 大连理工大学 | A kind of preparation method of the film modified nano graphene oxide pharmaceutical carrier of phosphatide for cancer target transmission |
| CN110856749A (en) * | 2018-08-23 | 2020-03-03 | 胡尚秀 | Boron-doped graphene quantum dot-containing nanocomposite and application thereof |
| CN110665486A (en) * | 2019-10-17 | 2020-01-10 | 福州大学 | Magnetic ferroferric oxide-PAMAM-antibody complex and preparation method and application thereof |
| CN110665486B (en) * | 2019-10-17 | 2021-05-18 | 福州大学 | Magnetic ferroferric oxide-PAMAM-antibody complex and preparation method and application thereof |
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