CN103043647A - Aminophenylboronic acid surface-modified nano-carbon material, as well as preparation method and application thereof - Google Patents
Aminophenylboronic acid surface-modified nano-carbon material, as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of inorganic materials and bioanalysis and in particular relates to an aminophenylboronic acid surface-modified nano-carbon material, as well as a preparation method and application thereof. According to the invention, an available nano-carbon material is subjected to carbodiimide reaction on the basis of acid treatment. Firstly, the surface of the nano-carbon material is modified by polylysine and polyethylene glycol/diglycolic acid, then the nano-carbon material reacts with aminophenylboronic acid for surface functional modification of the nano-carbon material. The synthesis method provided by the invention is simple and efficient. The prepared aminophenylboronic acid-functionalized nano-carbon material, as a nano-adsorbent, has a large specific surface area and strong selective specificity, exhibits better dispersibility and stability in aqueous solutions, and can rapidly achieve the effect of specifically separating and concentrating glycopeptide/glycoprotein. The material is significant in use value and application prospects in the glycoprotein proteomics field and the like.
Description
Technical field
The invention belongs to inorganic materials and bioassay technique field, be specifically related to nano-carbon material of finishing amino-benzene boric acid and its preparation method and application.
Background technology
The core of proteomics is on a large scale protein to be carried out analysis-by-synthesis, by the research to all protein character of certain species, individuality, organ, tissue or cell (comprising the dependency of interaction, protein and the disease of expression level, structure, distribution, function, abundance variation, posttranslational modification, thin inner cellular localization, protein-protein), make meticulous and accurately elaboration to protein function.Wherein the variation of the posttranslational modification protein Analysis and Identification that often has important physiological function thereby carry out posttranslational modification protein is bioanalysis field hot issue.
The functional protein of the glycoprotein combination that to be a class be formed by connecting with covalent linkage by carbohydrate and protein.Sugar chain not only changes the conformation of protein, and causes protein function to change.Glycosylation plays an important role, and participates in all more important biological processess such as receptor activation, signal transduction for folding, transportation, the location of protein.Simultaneously clinical biomarker and the target for the treatment of all are glycoprotein.Studies show that most sugars albumen is low-abundance protein, thereby very difficult being chosen by mass spectrum analyzed.In addition, the Ionization Efficiency of non-glycoprotein/non-glycopeptide is higher than glycoprotein/glycopeptide far away, and their a large amount of existence cause the signal of glycoprotein/glycopeptide in the complex system by severe inhibition, greatly reduce certified possibility.Therefore as prerequisite and the basis of analysis and identification glycoprotein, how sharp separation and enrichment glycoprotein more and more become the committed step of its detection of restriction and evaluation.
Present existing glycopeptide segment enriching method mainly contains following several: lectin affinity chromatography, Size Exclusion Chromatograph SEC method, hydrophilic chromatographic method, hydrazine hydrazone reaction method and boric acid concentration method.Recently the boric acid concentration method of development is based under the alkaline condition, boric acid can with the adjacent glycol of the cis of sugar chain in conjunction with forming stable covalent linkage, and can be reversibly with complete glycoprotein/glycopeptide wash-out under acidic conditions, so have widely suitability.In recent years, nano-carbon material with its development fast and application potential and more and more be applied to biological field.Therefore develop a kind of simply, the method for separation and concentration glycoprotein/glycopeptide becomes the important branch of proteomics research effectively.
Nano-carbon material is easy to functional modification take little, the good bio-compatibility of its particle diameter, high-specific surface area, surface characteristics as its method for separating and concentrating that is applied to trace peptide section in the proteome analysis provides may.
Summary of the invention
The object of the invention is to provide a kind of and is easy to synthesize, specificity is good, can carry out to glycoprotein/glycopeptide the nano-carbon material of the surface modified boric acid of separation and concentration.
The present invention also provides the preparation method of above-mentioned surface modified boric acid nano-carbon material.
The present invention further also provides the application of above-mentioned nano-carbon material.
The present invention be at original nano-carbon material on the basis of acidification, pass through Carbodiimide reaction, at first its surface is carried out the modification of poly-lysine (PL) and polyoxyethylene glycol diglycollic acid (PEG), react with amino-benzene boric acid (APBA) again, carry out the surface-functionalized modification of nano-carbon material, obtain the nano-carbon material of surface modified boric acid base group.
The present invention prepares the nano-carbon material of described surface modified boric acid base group by following method and step:
(1) nano-carbon material is carried out pre-treatment: the 1-5g nano-carbon material through ultra-sonic dispersion in 10-50mL H
2SO
4/ HNO
3(H
2SO
4: HNO
3=9:1-8:2(v/v)) in the solution, stirring at room 12-24 hour, be washed till neutrality with clear water; Place the 0.1-0.5M NaOH aqueous solution, in 90-95 ℃ of stirring 2-4 hour, clean with clear water; In 0.1-0.5M HCl solution, in 90-95 ℃ of stirring 2-4 hour, get acidifying/activation nano-carbon material after the clear water cleaning, drying; Above treating processes makes its surface produce carboxyl functional group;
(2) 0.5-1g acidifying/activation nano-carbon material is scattered in the 2-(N-morpholino that dissolves 5-10mL 60-80mg/mL N-maloyl imines) the ethyl sulfonic acid damping fluid; Ultrasonic wave was disperseed 2-5 minute, subsequently 5-10mL 60-80mg/mL N-ethyl dimethylamino-propyl carbodiimide hydrochloride is added fast, continue ultra-sonic dispersion 10-30min, reaction finishes, centrifugal collection nano-carbon material, supernatant discarded, and use the 2-(N-morpholino) the ethyl sulfonic acid damping fluid washes repeatedly;
(3) nano-carbon material with activation continues to be scattered in the 2-(N-morpholino of 8-10mL 2%-10% poly-lysine (Mw, 500-17,000)) in the ethyl sulfonic acid damping fluid, and fully DL vibration, reaction 3-12h; Question response finishes centrifugal collection, and cleans the nano-carbon material behind the polylysine modification;
(4) 0.2-1.0g polyoxyethylene glycol diglycollic acid is dissolved in the 2-(N-morpholino of 5-10mL 60-80mg/mL N-maloyl imines) the ethyl sulfonic acid damping fluid; Ultrasonic wave was disperseed 2-5 minute, subsequently 5-10mL 60-80mg/mL N-ethyl dimethylamino-propyl carbodiimide hydrochloride is added fast, continue ultrasonic 10-30min, polyoxyethylene glycol diglycollic acid after will activating subsequently and 1-2mL 10-20mg/mL amino-benzene boric acid add rapidly in the polylysine modification nano-carbon material, ultra-sonic dispersion, 4-10 ℃ of reaction spent the night 12-16 hour; Reaction finishes, and the nano-carbon material of centrifugal collection surface modified boric acid base group is dried after using ultrapure water fully to clean, and is for subsequent use.
In the step (1), described nano-carbon material can for Nano diamond, carbon nanotube, Graphene, fullerene-based material, be preferably Nano diamond.
In step (2)-(4), described 2-(N-morpholino) the ethyl sulfonic acid damping fluid can be phosphate buffered saline buffer.
In step (2) and (4), the 2-(N-morpholino of N-maloyl imines) ethyl sulfonic acid damping fluid, its concentration can be 100-200mM, and pH can be 6-6.5, such as being 6.1 etc.;
In the step (3), described poly-lysine is that molecular weight is at 5-15 ten thousand.
In the step (4), described polyoxyethylene glycol diglycollic acid is that molecular weight is at 200-2 ten thousand.
The nano-carbon material of the surface modified boric acid base group that the inventive method prepares has the structure of following formula:
The nano-carbon material synthetic method of surface modified boric acid base group of the present invention is simply effective, the functionalized nano carbon material of preparation has dispersed and stable preferably, it is large to have specific surface area, functional group is many, the characteristics of high specificity can be carried out high-sensitive specific isolation enrichment to the glycoprotein/glycopeptide in the biological specimen.
The nano-carbon material of surface modified boric acid base group of the present invention can be used for hanging down in the biological specimen specific isolation enrichment of abundance glycoprotein or glycopeptide, lays the foundation for expanding the application of nano-carbon material in biological field.This material has good practical value and application prospect in the bioanalysis field simultaneously.
Description of drawings
Fig. 1 is the synthetic method diagram of the nano-carbon material of surface modified boric acid base group.
Fig. 2 be the nano-carbon material of surface modified boric acid to the specific isolation concentration effect of glycopeptide in standard protein (Pp63 glycophosphoproteins) the peptide hydrolysis solution, contrast (a) specific enrichment not; (b) specific enrichment (numeral is designated as glycopeptide); (c) glycopeptide of specific enrichment is after Peptide N-glycosidase F removes sugar chain, and this material can specific isolation enrichment glycopeptide as can be known, and low abundance glycopeptide is had obvious concentration effect.
Fig. 3 be the nano-carbon material of surface modified boric acid under non-glycoprotein disturbs to standard protein (horseradish peroxidase) specific isolation concentration effect.Mixed protein solution S DS gel before contrast (a) enrichment separates; (b) sds gel of this material cleaning solution separates after the enrichment; (c) after the enrichment elute soln sds gel separation case of this material as can be known, glycoprotein obtains purifying after processing through the nano-carbon material of surface modified boric acid, proved invention material is to the specific isolation effect of sugar-protein in the mixed protein.
Embodiment
By following embodiment the separation and concentration analytic process that surface modified boric acid nano-carbon material provided by the present invention carries out glycopeptide, glycoprotein is described further.
Embodiment 1.Carry out the modification of amino-benzene boric acid on the Nano diamond surface:
(1) Nano diamond is carried out pre-treatment: the 1-5g Nano diamond through ultra-sonic dispersion in 10-50mL H
2SO
4/ HNO
3(9:1-8:2(v/v)) in the solution, stirring at room 12-24 hour, be washed till neutrality with clear water; Stirred 2-4 hour as for 90-95 ℃ of the 0.1-0.5M NaOH aqueous solution, clean with clear water; As in the 0.1-0.5M HCl solution 90-95 ℃ stirred 2-4 hour, after the clear water cleaning, drying acidifying/activation Nano diamond; Above treating processes makes its surface produce carboxyl functional group.
(2) 0.5-1g acidifying/activation Nano diamond is scattered in the 2-(N-morpholino of dissolving 5-10mL 60-80mg/mL N-maloyl imines) ethyl sulfonic acid damping fluid (100-200mM, pH 6.1).Ultrasonic wave was disperseed 2-5 minute, subsequently 5-10mL 60-80mg/mL N-ethyl dimethylamino-propyl carbodiimide hydrochloride is added fast, continue ultra-sonic dispersion 10-30min, reaction finishes, centrifugal collection Nano diamond, supernatant discarded, and use the 2-(N-morpholino) the ethyl sulfonic acid damping fluid washes repeatedly.
(3) Nano diamond with activation continues to be scattered in the 2-(N-morpholino of 8-10mL 2%-10% poly-lysine (Mw, 500-17,000)) in the ethyl sulfonic acid damping fluid and fully DL vibration, reaction 3-12h.Question response finishes centrifugal collection, and cleans the Nano diamond behind the polylysine modification.
(4) 0.2-1.0g polyoxyethylene glycol diglycollic acid is dissolved in the 2-(N-morpholino of 5-10mL 60-80mg/mL N-maloyl imines) ethyl sulfonic acid damping fluid (100-200mM pH 6.1).Ultrasonic wave was disperseed 2-5 minute, subsequently 5-10mL 60-80mg/mL N-ethyl dimethylamino-propyl carbodiimide hydrochloride is added fast, continue ultrasonic 10-30min, polyoxyethylene glycol diglycollic acid after will activating subsequently and 1-2mL 10-20mg/mL amino-benzene boric acid add rapidly in the polylysine modification Nano diamond, ultra-sonic dispersion, 4-10 ℃ of reaction spent the night 12-16 hour.Reaction finishes, and the Nano diamond of centrifugal collection surface modified boric acid base group is dried for subsequent use after using ultrapure water fully to clean.
Embodiment 2.Carry out the modification of amino-benzene boric acid on the Graphene surface:
(1) Graphene is carried out pre-treatment: the 1-5g Graphene through ultra-sonic dispersion in 10-50mL H
2SO
4/ HNO
3(9:1-8:2(v/v)) in the solution, stirring at room 12-24 hour, be washed till neutrality with clear water; Stirred 2-4 hour as for 90-95 ℃ of the 0.1-0.5M NaOH aqueous solution, clean with clear water; As in the 0.1-0.5M HCl solution 90-95 ℃ stirred 2-4 hour, after the clear water cleaning, drying graphene oxide; Above treating processes makes its surface produce carboxyl functional group.
(2) the 0.5-1g graphene oxide is scattered in the 2-(N-morpholino of dissolving 5-10mL 60-80mg/mL N-maloyl imines) ethyl sulfonic acid damping fluid (100-200mM, pH 6.1).Ultrasonic wave was disperseed 2-5 minute, subsequently 5-10mL 60-80mg/mL N-ethyl dimethylamino-propyl carbodiimide hydrochloride is added fast, continue ultra-sonic dispersion 10-30min, reaction finishes, the Graphene of centrifugal collection activation, supernatant discarded, and use the 2-(N-morpholino) the ethyl sulfonic acid damping fluid washes repeatedly.
(3) graphene oxide with activation continues to be scattered in the 2-(N-morpholino of 8-10mL 2%-10% poly-lysine (Mw, 500-17,000)) in the ethyl sulfonic acid damping fluid and fully DL vibration, reaction 3-12h.Question response finishes centrifugal collection, and cleans the Graphene behind the polylysine modification.
(4) 0.2-1.0g polyoxyethylene glycol diglycollic acid is dissolved in the 2-(N-morpholino of 5-10mL 60-80mg/mL N-maloyl imines) ethyl sulfonic acid damping fluid (100-200mM pH 6.1).Ultrasonic wave was disperseed 2-5 minute, subsequently 5-10mL 60-80mg/mL N-ethyl dimethylamino-propyl carbodiimide hydrochloride is added fast, continue ultrasonic 10-30min, polyoxyethylene glycol diglycollic acid after will activating subsequently and 1-2mL 10-20mg/mL amino-benzene boric acid add rapidly in the polylysine modification Graphene, ultra-sonic dispersion, 4-10 ℃ of reaction spent the night 12-16 hour.Reaction finishes, and the Graphene of centrifugal collection surface modified boric acid base group is dried for subsequent use after using ultrapure water fully to clean.
Embodiment 3.Carry out the modification of amino-benzene boric acid in carbon nano tube surface:
(1) carbon nanotube is carried out pre-treatment: the 1-5g carbon nanotube through ultra-sonic dispersion in 10-50mL H
2SO
4/ HNO
3(9:1-8:2(v/v)) in the solution, stirring at room 12-24 hour, be washed till neutrality with clear water; Stirred 2-4 hour as for 90-95 ℃ of the 0.1-0.5M NaOH aqueous solution, clean with clear water; As in the 0.1-0.5M HCl solution 90-95 ℃ stirred 2-4 hour, after the clear water cleaning, drying oxidation/acidifying carbon nanotube; Above treating processes makes its surface produce carboxyl functional group.
(2) 0.5-1g oxidation/acidifying carbon nanotube dispersed is in the 2-(N-morpholino of dissolving 5-10mL 60-80mg/mL N-maloyl imines) ethyl sulfonic acid damping fluid (100-200mM, pH 6.1).Ultrasonic wave was disperseed 2-5 minute, subsequently 5-10mL 60-80mg/mL N-ethyl dimethylamino-propyl carbodiimide hydrochloride is added fast, continue ultra-sonic dispersion 10-30min, reaction finishes, the carbon nanotube of centrifugal collection activation, supernatant discarded, and use the 2-(N-morpholino) the ethyl sulfonic acid damping fluid washes repeatedly.
(3) carbon nanotube with activation continues to be scattered in the 2-(N-morpholino of 8-10mL 2%-10% poly-lysine (Mw, 500-17,000)) in the ethyl sulfonic acid damping fluid and fully DL vibration, reaction 3-12h.Question response finishes centrifugal collection, and cleans the carbon nanotube behind the polylysine modification.
(4) 0.2-1.0g polyoxyethylene glycol diglycollic acid is dissolved in the 2-(N-morpholino of 5-10mL 60-80mg/mL N-maloyl imines) ethyl sulfonic acid damping fluid (100-200mM pH 6.1).Ultrasonic wave was disperseed 2-5 minute, subsequently 5-10mL 60-80mg/mL N-ethyl dimethylamino-propyl carbodiimide hydrochloride is added fast, continue ultrasonic 10-30min, polyoxyethylene glycol diglycollic acid after will activating subsequently and 1-2mL 10-20mg/mL amino-benzene boric acid add rapidly in the polylysine modification carbon nanotube, ultra-sonic dispersion, 4-10 ℃ of reaction spent the night 12-16 hour.Reaction finishes, and the carbon nanotube of centrifugal collection surface modified boric acid base group is dried for subsequent use after using ultrapure water fully to clean.
Embodiment 4. the surface modified boric acid Nano diamond is to the specific enrichment of glycopeptide in the Pp63 glycophosphoproteins matter peptide hydrolysis
(1) protein solution enzymolysis: get 1.0 mg Pp63 glycophosphoproteins standard substance, be dissolved in the 1.0 mL water, thermally denature 5min is with 1:40(enzyme/albumen) adding 25 μ g trypsinase.Hatching enzymolysis for 37 ℃ spends the night.Packing-80 ℃ saves backup.
(2) specific enrichment of glycopeptide: preparation Pp63 glycophosphoproteins pancreatin peptide hydrolysis in 1 mL50 mM ammonium bicarbonate soln (PH 8).In above-mentioned sample solution, add respectively the above-mentioned surface modified boric acid Nano diamond of 5 μ L, 10 mg/mL particle suspension liquid, incubated at room 1 hour.Hatch complete after, centrifugal 5 min under the 16 400 rpm rotating speeds remove supernatant.Then add 50 mM NH4HCO3 solution (PH 8) and repeatedly clean, more centrifugal removal supernatant.At last, add 50 μ L and contain 1% (v/v) trifluoroacetic acid aqueous solution, hatch 30 min under the room temperature, centrifugal collection supernatant elutriant.Simultaneously, hatch supernatant liquor and the scavenging solution of front are collected freeze-drying, heavily be dissolved in 5 μ L and contain 50% acetonitrile/0.1% (v/v) trifluoroacetic acid aqueous solution, carry out mass spectroscopy.
(3) MALDI-MS analyzes: analytical instrument adopts substance assistant laser desorpted ionized source-level Four bar ion trap/time-of-flight mass spectrometer (AXIMAQIT, Shimadzu).Get the glycopeptide enrichment elutriant of Pp63 glycophosphoproteins, concentrated supernatant liquor, each 1 μ L point target of concentrated cleaning solutions of hatching.Each sample spot adds 1 μ L, 10 mg/mL DHB (DHB, Sigma; Be dissolved in 50% v/v acetonitrile solution, include 0.1% (v/v) trifluoroacetic acid) as MALDI matrix.Instrument parameter is set as follows: MALDI source, 337 nm pulse of nitrogen laser apparatus, 3-5 nm FWHM); The collision gas of collision induced dissociation (CID), argon gas; The spectrogram accumulative total, 100; Laser intensity, fixed value.
Embodiment 5.The surface modified boric acid Nano diamond is to the specific enrichment of glycopeptide in horseradish peroxidase, bovine serum albumin, the myocardium protein mixed enzymolysis peptide section
(1) protein solution enzymolysis: get 1.0 mg protein standard substances, comprise bovine serum albumin, myocardium protein, horseradish peroxidase, be dissolved in respectively in the 1.0 mL water, thermally denature 5min is with 1:40(enzyme/albumen) adding 25 μ g trypsinase.Hatching enzymolysis for 37 ℃ spends the night.Packing-80 ℃ saves backup.
(2) specific enrichment of glycopeptide: in 1 mL, 50 mM ammonium bicarbonate solns (PH 8), prepare respectively horseradish peroxidase peptide section, the bovine serum albumin peptide section of different ratios, the mixture of myocardium protein peptide section.In above-mentioned sample solution, add respectively the above-mentioned surface modified boric acid Nano diamond of 5 μ L, 10 mg/mL particle suspension liquid, incubated at room 1 hour.Hatch complete after, centrifugal 5 min under the 16 400 rpm rotating speeds remove supernatant.Then add 50 mM ammonium bicarbonate solns (PH 8) and repeatedly clean, more centrifugal removal supernatant.At last, add 50 μ L and contain 1% (v/v) trifluoroacetic acid aqueous solution, hatch 30 min under the room temperature, centrifugal collection supernatant elutriant.Simultaneously, hatch supernatant liquor and the scavenging solution of front are collected freeze-drying, heavily be dissolved in 5 μ L and contain 50% acetonitrile/0.1% (v/v) trifluoroacetic acid aqueous solution, carry out mass spectroscopy.
(3) MALDI-MS analyzes: analytical instrument adopts substance assistant laser desorpted ionized source-level Four bar ion trap/time-of-flight mass spectrometer (AXIMAQIT, Shimadzu).The accurate glycoprotein sample of label taking glycopeptide enrichment elutriant, concentrated supernatant liquor, each 1 μ L point target of concentrated cleaning solutions of hatching.Each sample spot adds 1 μ L, 10 mg/mL DHB (DHB, Sigma; Be dissolved in 50% v/v acetonitrile solution, include 0.1% (v/v) trifluoroacetic acid) as MALDI matrix.Instrument parameter is set as follows: MALDI source, 337 nm pulse of nitrogen laser apparatus, 3-5 nm FWHM); The collision gas of collision induced dissociation (CID), argon gas; The spectrogram accumulative total, 100; Laser intensity, fixed value.
Embodiment 6.The surface modified boric acid Nano diamond is used for the specific enrichment of horseradish peroxidase, bovine serum albumin, myocardium protein mixed protein glycoprotein
Get and contain horseradish peroxidase, bovine serum albumin, myocardium protein, concentration is all 50 ng/ μ L, and volume is 200 μ L.Add the surface modified boric acid Nano diamond material that 100 μ L concentration are 10 mg/mL, whole system PH value is about 8.5, room temperature 30 min that vibrate.Centrifugal collection, employing PH is 8.5 Tris-HCl buffer solution for cleaning material twice, each 200 μ L; In material, add 50% v/v acetonitrile solution, include 0.1% (v/v) trifluoroacetic acid solution with the glycoprotein of enrichment wash-out from the material, 15 min that vibrate dye the laggard line scanning of colour developing and analyze carrying out 12%SDS gel electrophoresis separation, silver after scavenging solution, the elutriant freeze-drying.
Claims (8)
1. the preparation method of the nano-carbon material of a surface modified boric acid base group is characterized in that concrete steps are:
(1) nano-carbon material is carried out pre-treatment: the 1-5g nano-carbon material through ultra-sonic dispersion in 10-50mL H
2SO
4/ HNO
3In the solution, stirring at room 12-24 hour, be washed till neutrality with clear water, H
2SO
4/ HNO
3H in the solution
2SO
4: HNO
3=9:1-8:2(v/v); Place the 0.1-0.5M NaOH aqueous solution, in 90-95 ℃ of stirring 2-4 hour, clean with clear water; Place 0.1-0.5M HCl solution, in 90-95 ℃ of stirring 2-4 hour, get acidifying/activation nano-carbon material after the clear water cleaning, drying; Above treating processes makes its surface produce carboxyl functional group;
(2) 0.5-1g acidifying/activation nano-carbon material is scattered in the 2-(N-morpholino that dissolves 5-10mL 60-80mg/mL N-maloyl imines) the ethyl sulfonic acid damping fluid; Ultrasonic wave was disperseed 2-5 minute, subsequently 5-10mL 60-80mg/mL N-ethyl dimethylamino-propyl carbodiimide hydrochloride is added fast, continue ultra-sonic dispersion 10-30min, reaction finishes, centrifugal collection nano-carbon material, supernatant discarded, and use the 2-(N-morpholino) the ethyl sulfonic acid damping fluid washes repeatedly;
(3) nano-carbon material with activation continues to be scattered in the 2-(N-morpholino of 8-10mL 2%-10% poly-lysine) in the ethyl sulfonic acid damping fluid, and fully DL vibration, reaction 3-12h; Question response finishes centrifugal collection, and cleans the nano-carbon material behind the polylysine modification;
(4) 0.2-1.0g polyoxyethylene glycol diglycollic acid is dissolved in the 2-(N-morpholino of 5-10mL 60-80mg/mL N-maloyl imines) the ethyl sulfonic acid damping fluid; Ultrasonic wave was disperseed 2-5 minute, subsequently 5-10mL 60-80mg/mL N-ethyl dimethylamino-propyl carbodiimide hydrochloride is added fast, continue ultrasonic 10-30min, polyoxyethylene glycol diglycollic acid after will activating subsequently and 1-2mL 10-20mg/mL amino-benzene boric acid add rapidly in the polylysine modification nano-carbon material, ultra-sonic dispersion, 4-10 ℃ of reaction spent the night 12-16 hour; Reaction finishes, and the nano-carbon material of centrifugal collection surface modified boric acid base group is dried after using ultrapure water fully to clean.
2. the preparation method of the nano-carbon material of surface modified boric acid base group according to claim 1 is characterized in that in the step (1), described nano-carbon material is Nano diamond, carbon nanotube, Graphene or fullerene-based material.
3. the preparation method of the nano-carbon material of surface modified boric acid base group according to claim 1 is characterized in that in step (2)-(4) described 2-(N-morpholino) the ethyl sulfonic acid damping fluid is phosphate buffered saline buffer.
4. the preparation method of the nano-carbon material of surface modified boric acid base group according to claim 1, it is characterized in that in step (2) and (4), the 2-(N-morpholino of N-maloyl imines) ethyl sulfonic acid damping fluid, its concentration is 100-200mM, pH is 6-6.5.
5. the preparation method of the nano-carbon material of surface modified boric acid base group according to claim 1 is characterized in that in the step (3), and described poly-lysine is that molecular weight is at 5-15 ten thousand.
6. the preparation method of the nano-carbon material of surface modified boric acid base group according to claim 1 is characterized in that in the step (4), and described polyoxyethylene glycol diglycollic acid is that molecular weight is at 200-2 ten thousand.
7. the nano-carbon material of the surface modified boric acid base group that is prepared by the described method of one of claim 1-6 has the structure of following formula:
8. the purposes of surface modified boric acid nano-carbon material claimed in claim 1 in preparation specific isolation enrichment glycoprotein and glycopeptide.
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| CN104374848A (en) * | 2013-08-14 | 2015-02-25 | 中国科学院大连化学物理研究所 | Method for enriching glycopeptide by phenylboronic acid material |
| CN105223248A (en) * | 2015-09-21 | 2016-01-06 | 安徽师范大学 | Based on phenyl boric acid imprinted polymer/carbon nano tube modified electrode and its preparation method and application |
| CN105358481A (en) * | 2013-07-12 | 2016-02-24 | 独立行政法人产业技术综合研究所 | Carbon-containing solid acid having sulfonate group |
| CN108927116A (en) * | 2018-07-20 | 2018-12-04 | 大连医科大学 | A kind of preparation method for the stannic oxide/graphene nano material that phenyl boric acid functionalization is chitosan-modified |
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| CN112574431A (en) * | 2021-02-26 | 2021-03-30 | 中国农业科学院农业质量标准与检测技术研究所 | Phenylboronic acid functionalized multi-walled carbon nanotube magnetic nanocomposite and preparation method and application thereof |
| CN114644734A (en) * | 2020-12-02 | 2022-06-21 | 中国科学院大连化学物理研究所 | Glycopeptide-enriched polymer material, preparation and application |
| WO2022260136A1 (en) * | 2021-06-11 | 2022-12-15 | 国立大学法人京都大学 | Surface-modified nanoparticle |
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