CN103333295A - Preparation method of thymopentin molecularly-imprinted magnetic microspheres - Google Patents
Preparation method of thymopentin molecularly-imprinted magnetic microspheres Download PDFInfo
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Abstract
The invention provides a preparation method of thymopentin molecularly-imprinted magnetic microspheres, which comprises the following steps: preparing oleic-acid-modified Fe3O4 magnetic nanoparticles, and preparing carboxylated Fe3O4 magnetic nanoparticles with methyl methacrylate and methacrylic acid by using sodium lauryl sulfate as a surfactant, potassium persulfate as a initiator and ethylene glycol dimethacrylate as a crosslinking agent; and finally, successfully preparing the thymopentin molecularly-imprinted magnetic microspheres by using thymopentin as a template molecule, acrylamide as a functional monomer, ethylene glycol dimethacrylate as a crosslinking agent and carboxylated Fe3O4 as a magnetic carrier. The preparation method has the advantages of mild reaction conditions, high imprinting efficiency, controllable magnetic microsphere size, high adsorption capacity and easy separation. By integrating the single recognition characteristic of the molecularly-imprinted polymer and the easy separation characteristic of the magnetic microspheres under the action of an external magnetic field, the preparation method has wide application prospects in the fields of separation, enrichment, detection, medicine targeted therapy and the like.
Description
Technical field
The invention belongs to the preparation method of peptide molecule trace magnetic microsphere, be specially the preparation method of thymopeptide-5 molecular blotting magnetic microsphere, belong to the preparing technical field of functional materials.
Background technology
Fields such as biology, medical science studies show that requisite regulatory factor all is less peptide class in many physiological processs.Along with the increase to AIDS, cancer class pharmaceutical requirements, in the world the research speed of peptide medicament and peptide class reagent was accelerated in recent years, the biological extraction peptide can not satisfy the demand of market and medicament research and development, and therefore the synthetic peptide technology has obtained developing rapidly.But the synthetic peptide technology is because its special synthesis strategy has brought certain difficulty for the product separation purifying.
Thymopoietin (Thymopoietin, TP) be isolated a kind of 49 amino acid whose polypeptide from people's thymic tissue, relative molecular mass is 5562 single polypeptide compound, it has the thymocyte of promotion and periphery T cell and B cytodifferentiation grows, and regulates biological activitys such as body's immunity.What wherein, play immunologic function is the 32-36 amino acids of TP.(Arg-Lys-Asp-Va1-Tyr TP5) is the pentapeptide of synthetic to thymopeptide-5, and the 32-36 amino acids of its amino-acid sequence and structure and TP is formed identical, and biologic activity is identical.
In the process of synthetic thymopeptide-5, along with the prolongation of peptide chain, be linked at the active amino acid side chain on the peptide chain, if do not protected (sealing), will in reaction, produce a series of by product, product purification is brought difficulty.At present, utilize the thick peptide of solid phase synthesis technique preparation to be mostly to adopt inverse technique (RP-HPLC) to separate, this Technology Need carries out under condition of high voltage, and this must cause, and equipment energy consumption increases, cost improves.Therefore, searching is a kind of efficiently, purification process has huge economic cheaply.
Molecular imprinting is the effective and inexpensive method for preparing the polymer materials with special identification function fast.Has affinity and selectivity is good based on the imprinted polymer of this technology preparation, advantages such as the anti-adverse environment ability is strong, good stability, long service life and applied range, be widely used in chemical catalysis, sepn process (comprises film analysis and Solid-Phase Extraction, and chromatographic separation), Clinical Application Analysis (comprises slowly released and controlled-drug delivery system, the SCL administration, pharmaceutical carrier, activity is regulated drug delivery system, targeting drug delivery system and capture systems), sensing technology fields such as (comprising electrochemical sensor, Optochemical sensor and mass-sensitive sensor).
The method that adopts molecular imprinting to combine with magnetic microsphere prepares thymopeptide-5 molecular engram polymer magnetic microballoon, can make molecular engram microsphere have magnetic and specific recognition capability concurrently, distinctive " memory " reaches specific recognition to the thymopeptide-5 molecular structure by this compound, simultaneously can reach sharp separation again adding under the action of a magnetic field, greatly improve the separation efficiency of thymopeptide-5.
At present, the engram technology of biological micromolecules such as amino acid and derivative thereof, sugar, steroid, Nucleotide, hormone, VITAMIN is comparatively ripe, the investigator just is being devoted to molecular imprinting is applied to biomacromolecule, and the biomacromolecule that successfully is used for molecular imprinting has ribonuclease A, Fibrinogen, N,O-Diacetylmuramidase, glutamine synthetase, streptavidin, urease, albumin, myosin, oxyphorase, cytochrome C, lactoperoxidase, horseradish peroxidase, glucose oxidase, Uteracon, antidiuretic hormone.By literature search, do not find as yet to adopt molecular imprinting to prepare the relevant report of thymopeptide-5 molecular blotting magnetic microsphere.
Summary of the invention
The technical problem that solves
For fear of the deficiencies in the prior art part, the present invention proposes a kind of preparation method of thymopeptide-5 molecular blotting magnetic microsphere.
Technical scheme
A kind of preparation method of thymopeptide-5 molecular blotting magnetic microsphere is characterized in that step is as follows:
Step 1: according to mol ratio 1:2 with FeCl
24H
2O and FeCl
36H
2O is dissolved in distilled water, mixes under 30 ℃, nitrogen protection and stirs, and the NaOH solution that drips 1mol/L then makes pH reach 11, obtains the black turbid solution; Adopt hydrochloric acid black turbid solution pH=7, add the oleic acid of distilled water volume 5%, be warming up to 80 ℃, at 800rmpmin
-1Behind the mechanical stirring 2h, isolate the black solid material under the outside magnetic field effect under the rotating speed; Adopt distilled water and dehydrated alcohol to intersect washing repeatedly, at 30 ℃ of following vacuum-drying 24h, obtain the Fe of oleic acid modification
3O
4Magnetic nano-particle;
Step 2: with the Fe of oleic acid modification
3O
4Magnetic nano-particle adds ultra-sonic dispersion in the normal hexane according to 0.5g/mL, forms the oil base magnetic fluid;
Step 3: be the lauryl sodium sulfate aqueous solution of 3.5mg/mL with concentration, join in the oil base magnetic fluid, form magnetic template emulsion, add methyl methacrylate MMA and methacrylic acid MAA more successively, mechanical stirring 2h under 60～70 ℃, 800r/min rotating speed, the Potassium Persulphate KPS, the Ethylene glycol dimethacrylate EGDMA of 0.4～0.6mL that continue to add 0.01g, thermal-initiated polymerization 20h obtains the Fe of carboxyl-functional
3O
4Magnetic nano-particle; The volume ratio of described methyl methacrylate MMA and methacrylic acid MAA is 2:1～4:1;
Step 4: thymopeptide-5 TP-5 and acrylamide AM are dissolved in the distilled water, behind 25 ℃ of following prepolymerization 20～60min, add the Fe of 1.2% carboxyl-functional of this distilled water quality
3O
4Magnetic nano-particle, the ultrasonic 15min of room temperature forms the water mixed solution; The mol ratio of described thymopeptide-5 TP-5 and acrylamide AM is 1:20～1:28;
Step 5: under the 800r/min stirring action, the potassium persulfate solution 1mL that adds EGDMA, the 1.5mg/mL of 0.3～0.5mL in the water mixed solution successively, in 30 ℃ of water-baths behind thermal-initiated polymerization 5～7h, be cooled to room temperature, adopt distilled water and dehydrated alcohol to intersect washing repeatedly, at 30 ℃ of following vacuum-drying 24h, obtain molecular blotting magnetic microsphere;
Step 6: molecular blotting magnetic microsphere is joined in the mixing solutions of phosphate buffered saline buffer and methyl alcohol, at 25 ℃, 120rmpmin
-1Shaking table vibration 10h is with wash-out thymopeptide-5 TP-5 under the rotating speed; Again with distilled water wash repeatedly, at 25 ℃ of following vacuum-drying 24h, obtain the thymopeptide-5 molecular blotting magnetic microsphere; The volume ratio of described phosphate buffered saline buffer and methyl alcohol is 60:40.
The pH=7 of described phosphate buffered saline buffer.
Adopting distilled water and dehydrated alcohol to intersect in the step 1 washs 6 times.
Step 5 adopts distilled water and dehydrated alcohol to intersect and washs 3 times.
Step 6 distilled water wash 6 times.
Beneficial effect
The present invention proposes a kind of preparation method of thymopeptide-5 molecular blotting magnetic microsphere.The Fe that at first prepares the oleic acid modification
3O
4Magnetic nano-particle under sodium lauryl sulphate is made tensio-active agent, is initiator with the Potassium Persulphate then, and Ethylene glycol dimethacrylate is linking agent, utilizes methyl methacrylate and methacrylic acid to prepare the Fe of carboxyl-functional
3O
4Magnetic nano-particle.Last is that template molecule, acrylamide are that function monomer, Ethylene glycol dimethacrylate are the Fe of linking agent, carboxyl-functional with the thymopeptide-5
3O
4Be magnetic carrier, successfully prepare the thymopeptide-5 molecular blotting magnetic microsphere.Preparation method's reaction conditions gentleness of the present invention, trace efficient height, the magnetic microsphere particle diameter is controlled, and adsorptive capacity is high and easily separated.This preparation method unites two into one the single-minded evident characteristics of molecularly imprinted polymer with the characteristic that magnetic microsphere easily separates under extraneous the action of a magnetic field, therefore have a wide range of applications in fields such as separation, enrichment, detection, drug targeting treatments.
Embodiment
Now in conjunction with the embodiments the present invention is further described:
Embodiment 1: with 0.01moL FeCl
24H
2O and 0.02moLFeCl
36H
2O is dissolved in 30mL distilled water, drips 1mol/L NaOH solution behind the mechanical stirring 1h to pH value of solution=11 under 30 ℃, nitrogen protection, continues to stir 2h, gets the black turbid solution.Transfer this pH value of solution=7 with hydrochloric acid, add 1.5mL oleic acid, be warming up to 80 ℃, 800rmpmin
-1Behind the mechanical stirring 2h, isolate the black solid material under the outside magnetic field effect under the rotating speed.Successively with distilled water, each 40mL washing of dehydrated alcohol 6 times, 30 ℃ of following vacuum-drying 24h get the Fe of oleic acid modification with it
3O
4Magnetic nano-particle.Take by weighing the Fe of 0.6g oleic acid modification
3O
4The magnetic nano-particle ultra-sonic dispersion forms the oil base magnetic fluid in the 1.2mL normal hexane.Configuration concentration is the lauryl sodium sulfate aqueous solution 20mL of 3.5mg/mL, changes it over to oil base magnetic fluid, forms magnetic template emulsion, adds 0.9mLMMA and 0.3mLMAA successively, at 70 ℃, 800rmpmin
-12h under the following mechanical stirring continue to add 0.01gKPS, 0.6mLEGDMA, and thermal-initiated polymerization 20h obtains the Fe of carboxyl-functional
3O
4Magnetic nano-particle.0.029gTP-5 and 1mmoLAM are dissolved in respectively in the 10mL distilled water, behind 25 ℃ of following prepolymerization 40min.The Fe that adds the 0.12g carboxyl-functional
3O
4Magnetic nano-particle, the ultrasonic 15min of room temperature is at 800rmpmin
-1Under the stirring action, add 0.4mLEGDMA, 1.5mg/mL potassium persulfate solution 1mL successively, in 30 ℃ of following water-baths behind the thermal-initiated polymerization 6h.Be cooled to room temperature, with distilled water, each 40mL washing of ethanol 3 times, 30 ℃ of following vacuum-drying 24h get molecular blotting magnetic microsphere successively.
Molecular blotting magnetic microsphere is added in the mixing solutions of 50mL phosphate buffered saline buffer (pH=7) and methyl alcohol, both amount ratios are 60:40 (volume ratio).At 25 ℃, 120rmpmin
-1Shaking table vibration 10h wash-out TP-5 under the rotating speed.With 40mL distilled water wash 6 times of molecular blotting magnetic microsphere behind the wash-out.25 ℃ of following vacuum-drying 24h obtain the thymopeptide-5 molecular blotting magnetic microsphere.
After testing, prepared thymopeptide-5 molecular blotting magnetic microsphere median size is 2.6 μ m.Saturated extent of adsorption is 15.63mg/g, and the trace factor is 1.46.
Embodiment 2: with 0.01moL FeCl
24H
2O and 0.02moLFeCl
36H
2O is dissolved in 30mL distilled water, drips 1mol/L NaOH solution behind the mechanical stirring 1h to pH value of solution=11 under 30 ℃, nitrogen protection, continues to stir 2h, gets the black turbid solution.Transfer this pH value of solution=7 with hydrochloric acid, add 1.5mL oleic acid, be warming up to 80 ℃, 800rmpmin
-1Behind the mechanical stirring 2h, isolate the black solid material under the outside magnetic field effect under the rotating speed.Successively with distilled water, each 40mL washing of dehydrated alcohol 6 times, 30 ℃ of following vacuum-drying 24h get the Fe of oleic acid modification with it
3O
4Magnetic nano-particle.Take by weighing the Fe of 0.6g oleic acid modification
3O
4The magnetic nano-particle ultra-sonic dispersion forms the oil base magnetic fluid in the 1.2mL normal hexane, configuration concentration is the lauryl sodium sulfate aqueous solution 20mL of 3.5mg/mL, change it over to oil base magnetic fluid, form magnetic template emulsion, add 0.8mLMMA and 0.2mLMAA successively, at 60 ℃, 800rmpmin
-12h under the following mechanical stirring continue to add 0.01gKPS, 0.4mLEGDMA, and thermal-initiated polymerization 20h obtains the Fe of carboxyl-functional
3O
4Magnetic nano-particle.0.024gTP-5 and 1mmoLAM are dissolved in respectively in the 10mL distilled water, behind 25 ℃ of following prepolymerization 60min.The Fe that adds the 0.12g carboxyl-functional
3O
4Magnetic nano-particle, the ultrasonic 15min of room temperature is at 800rmpmin
-1Under the stirring action, add 0.5mLEGDMA, 1.5mg/mL potassium persulfate solution 1mL successively, in 30 ℃ of following water-baths behind the thermal-initiated polymerization 7h.Be cooled to room temperature, with distilled water, each 40mL washing of ethanol 3 times, 30 ℃ of following vacuum-drying 24h get molecular blotting magnetic microsphere successively.
Molecular blotting magnetic microsphere is added in the mixing solutions of 50mL phosphate buffered saline buffer (pH=7) and methyl alcohol, both amount ratios are 60:40 (volume ratio).At 25 ℃, 120rmpmin
-1Shaking table vibration 10h wash-out TP-5 under the rotating speed.With 40mL distilled water wash 6 times of molecular blotting magnetic microsphere behind the wash-out.25 ℃ of following vacuum-drying 24h obtain the thymopeptide-5 molecular blotting magnetic microsphere.
After testing, prepared thymopeptide-5 molecular blotting magnetic microsphere median size is 2.3 μ m.Saturated extent of adsorption is 15.5mg/g, and the trace factor is 1.37.
Embodiment 3: with 0.01moL FeCl
24H
2O and 0.02moLFeCl
36H
2O is dissolved in 30mL distilled water, drips 1mol/L NaOH solution behind the mechanical stirring 1h to pH value of solution=11 under 30 ℃, nitrogen protection, continues to stir 2h, gets the black turbid solution.Transfer this pH value of solution=7 with hydrochloric acid, add 1.5mL oleic acid, be warming up to 80 ℃, 800rmpmin
-1Behind the mechanical stirring 2h, isolate the black solid material under the outside magnetic field effect under the rotating speed.Successively with distilled water, each 40mL washing of dehydrated alcohol 6 times, 30 ℃ of following vacuum-drying 24h get the Fe of oleic acid modification with it
3O
4Magnetic nano-particle.Take by weighing the Fe of 0.6g oleic acid modification
3O
4The magnetic nano-particle ultra-sonic dispersion forms the oil base magnetic fluid in the 1.2mL normal hexane, configuration concentration is the lauryl sodium sulfate aqueous solution 20mL of 3.5mg/mL, change it over to oil base magnetic fluid, form magnetic template emulsion, add 0.8mLMMA and 0.4mLMAA successively, at 65 ℃, 800rmpmin
-12h under the following mechanical stirring continue to add 0.01gKPS, 0.5mLEGDMA, and thermal-initiated polymerization 20h obtains the Fe of carboxyl-functional
3O
4Magnetic nano-particle.0.029gTP-5 and 1mmoLAM are dissolved in respectively in the 10mL distilled water, behind 25 ℃ of following prepolymerization 40min.The Fe that adds the 0.12g carboxyl-functional
3O
4Magnetic nano-particle, the ultrasonic 15min of room temperature is at 800rmpmin
-1Under the mechanical agitation, add 0.4mLEGDMA, 1.5mg/mL potassium persulfate solution 1mL successively, in 30 ℃ of water-baths behind the thermal-initiated polymerization 6h.Be cooled to room temperature, with distilled water, each 40mL washing of ethanol 3 times, 30 ℃ of following vacuum-drying 24h get molecular blotting magnetic microsphere successively.
Molecular blotting magnetic microsphere is added in the mixing solutions of 50mL phosphate buffered saline buffer (pH=7) and methyl alcohol, both amount ratios are 60:40 (volume ratio).At 25 ℃, 120rmpmin
-1Shaking table vibration 10h wash-out TP-5 under the rotating speed.With 40mL distilled water wash 6 times of molecular blotting magnetic microsphere behind the wash-out.25 ℃ of following vacuum-drying 24h obtain the thymopeptide-5 molecular blotting magnetic microsphere.
After testing, prepared thymopeptide-5 molecular blotting magnetic microsphere median size is 2.8 μ m.Saturated extent of adsorption is 15.52mg/g, and the trace factor is 1.42.
Embodiment 4: with 0.01moL FeCl
24H
2O and 0.02moLFeCl
36H
2O is dissolved in 30mL distilled water, drips 1mol/L NaOH solution behind the mechanical stirring 1h to solution PH=11 under 30 ℃, nitrogen protection, continues to stir 2h, gets the black turbid solution.Transfer this solution PH=7 with hydrochloric acid, add 1.5mL oleic acid, be warming up to 80 ℃, 800rmpmin
-1Behind the mechanical stirring 2h, isolate the black solid material under the outside magnetic field effect under the rotating speed.Successively with distilled water, each 40mL washing of dehydrated alcohol 6 times, 30 ℃ of following vacuum-drying 24h get the Fe of oleic acid modification with it
3O
4Magnetic nano-particle.Take by weighing the Fe of 0.6g oleic acid modification
3O
4The magnetic nano-particle ultra-sonic dispersion forms the oil base magnetic fluid in the 1.2mL normal hexane, configuration concentration is the lauryl sodium sulfate aqueous solution 20mL of 3.5mg/mL, change it over to oil base magnetic fluid, form magnetic template emulsion, add 0.8mLMMA and 0.2mLMAA successively, at 60 ℃, 800rmpmin
-12h under the following mechanical stirring continue to add 0.01gKPS, 0.4mLEGDMA, and thermal-initiated polymerization 20h obtains the Fe of carboxyl-functional
3O
4Magnetic nano-particle.0.029gTP-5 and 1mmoLAM are dissolved in respectively in the 10mL distilled water, behind 25 ℃ of following prepolymerization 40min.The Fe that adds the 0.12g carboxyl-functional
3O
4Magnetic nano-particle, the ultrasonic 15min of room temperature is at 800rmpmin
-1Under the stirring action, add 0.4mLEGDMA, 1.5mg/mL potassium persulfate solution 1mL successively, in 30 ℃ of water-baths behind the thermal-initiated polymerization 6h.Be cooled to room temperature, with distilled water, each 40mL washing of ethanol 3 times, 30 ℃ of following vacuum-drying 24h get molecular blotting magnetic microsphere successively.
Molecular blotting magnetic microsphere is added in the mixing solutions of 50mL phosphate buffered saline buffer (pH=7) and methyl alcohol, both amount ratios are 60:40 (volume ratio).At 25 ℃, 120rmpmin
-1Shaking table vibration 10h wash-out TP-5 under the rotating speed.With 40mL distilled water wash 6 times of molecular blotting magnetic microsphere behind the wash-out.25 ℃ of following vacuum-drying 24h obtain the thymopeptide-5 molecular blotting magnetic microsphere.
After testing, prepared thymopeptide-5 molecular blotting magnetic microsphere median size is 2.6 μ m.Saturated extent of adsorption is 14.1mg/g, and the trace factor is 1.35.
Embodiment 5: with 0.01moL FeCl
24H
2O and 0.02moLFeCl
36H
2O is dissolved in 30mL distilled water, drips 1mol/L NaOH solution behind the mechanical stirring 1h to pH value of solution=11 under 30 ℃, nitrogen protection, continues to stir 2h, gets the black turbid solution.Transfer this pH value of solution=7 with hydrochloric acid, add 1.5mL oleic acid, be warming up to 80 ℃, 800rmpmin
-1Behind the mechanical stirring 2h, isolate the black solid material under the outside magnetic field effect under the rotating speed.Successively with distilled water, each 40mL washing of dehydrated alcohol 6 times, 30 ℃ of following vacuum-drying 24h get the Fe of oleic acid modification with it
3O
4Magnetic nano-particle.Take by weighing the Fe of 0.6g oleic acid modification
3O
4The magnetic nano-particle ultra-sonic dispersion forms the oil base magnetic fluid in the 1.2mL normal hexane, configuration concentration is the lauryl sodium sulfate aqueous solution 20mL of 3.5mg/mL, change it over to oil base magnetic fluid, form magnetic template emulsion, add 0.9mLMMA and 0.3mLMAA successively, at 70 ℃, 800rmpmin
-12h under the following mechanical stirring continue to add 0.01gKPS, 0.6mLEGDMA, and thermal-initiated polymerization 20h obtains the Fe of carboxyl-functional
3O
4Magnetic nano-particle.0.033gTP-5 and 1mmoLAM are dissolved in respectively in the 10mL distilled water, behind 25 ℃ of following prepolymerization 20min.The Fe that adds the 0.12g carboxyl-functional
3O
4Magnetic nano-particle, the ultrasonic 15min of room temperature is at 800rmpmin
-1Under the stirring action, add 0.3mLEGDMA, 1.5mg/mL potassium persulfate solution 1mL successively, in 30 ℃ of following water-baths behind the thermal-initiated polymerization 5h.Be cooled to room temperature, with distilled water, each 40mL washing of ethanol 3 times, 30 ℃ of following vacuum-drying 24h get molecular blotting magnetic microsphere successively.
Molecular blotting magnetic microsphere is added in the mixing solutions of 50mL phosphate buffered saline buffer (pH=7) and methyl alcohol, both amount ratios are 60:40 (volume ratio).At 25 ℃, 120rmpmin
-1Shaking table vibration 10h wash-out TP-5 under the rotating speed.With 40mL distilled water wash 6 times of molecular blotting magnetic microsphere behind the wash-out.25 ℃ of following vacuum-drying 24h obtain the thymopeptide-5 molecular blotting magnetic microsphere.
After testing, prepared thymopeptide-5 molecular blotting magnetic microsphere median size is 2.2 μ m.Saturated extent of adsorption is 12.6mg/g, and the trace factor is 1.32.
Embodiment 6: with 0.01moL FeCl
24H
2O and 0.02moLFeCl
36H
2O is dissolved in 30mL distilled water, drips 1mol/L NaOH solution behind the mechanical stirring 1h to pH value of solution=11 under 30 ℃, nitrogen protection, continues to stir 2h, gets the black turbid solution.Transfer this pH value of solution=7 with hydrochloric acid, add 1.5mL oleic acid, be warming up to 80 ℃, 800rmpmin
-1Behind the mechanical stirring 2h, isolate the black solid material under the outside magnetic field effect under the rotating speed.Successively with distilled water, each 40mL washing of dehydrated alcohol 6 times, 30 ℃ of following vacuum-drying 24h get the Fe of oleic acid modification with it
3O
4Magnetic nano-particle.Take by weighing the Fe of 0.6g oleic acid modification
3O
4The magnetic nano-particle ultra-sonic dispersion forms the oil base magnetic fluid in the 1.2mL normal hexane, configuration concentration is the lauryl sodium sulfate aqueous solution 20mL of 3.5mg/mL, change it over to oil base magnetic fluid, form magnetic template emulsion, add 0.9mLMMA and 0.3mLMAA successively, at 70 ℃, 800rmpmin
-12h under the following mechanical stirring continue to add 0.01gKPS, 0.6mLEGDMA, and thermal-initiated polymerization 20h obtains the Fe of carboxyl-functional
3O
4Magnetic nano-particle.0.024gTP-5 and 1mmoLAM are dissolved in respectively in the 10mL distilled water, behind 25 ℃ of following prepolymerization 60min.The Fe that adds the 0.12g carboxyl-functional
3O
4Magnetic nano-particle, the ultrasonic 15min of room temperature is at 800rmpmin
-1Under the stirring action, add 0.5mLEGDMA, 1.5mg/mL potassium persulfate solution 1mL successively, in 30 ℃ of following water-baths behind the thermal-initiated polymerization 7h.Be cooled to room temperature, with distilled water, each 40mL washing of ethanol 3 times, 30 ℃ of following vacuum-drying 24h get molecular blotting magnetic microsphere successively.
Molecular blotting magnetic microsphere is added in the mixing solutions of 50mL phosphate buffered saline buffer (pH=7) and methyl alcohol, both amount ratios are 60:40 (volume ratio).At 25 ℃, 120rmpmin
-1Shaking table vibration 10h wash-out TP-5 under the rotating speed.With 40mL distilled water wash 6 times of molecular blotting magnetic microsphere behind the wash-out.25 ℃ of following vacuum-drying 24h obtain the thymopeptide-5 molecular blotting magnetic microsphere.
After testing, prepared thymopeptide-5 molecular blotting magnetic microsphere median size is 2.5 μ m.Saturated extent of adsorption is 15.2mg/g, and the trace factor is 1.36.
Claims (5)
1. the preparation method of a thymopeptide-5 molecular blotting magnetic microsphere is characterized in that step is as follows:
Step 1: according to mol ratio 1:2 with FeCl
24H
2O and FeCl
36H
2O is dissolved in distilled water, mixes under 30 ℃, nitrogen protection and stirs, and the NaOH solution that drips 1mol/L then makes pH reach 11, obtains the black turbid solution; Adopt hydrochloric acid black turbid solution pH=7, add the oleic acid of distilled water volume 5%, be warming up to 80 ℃, at 800rmpmin
-1Behind the mechanical stirring 2h, isolate the black solid material under the outside magnetic field effect under the rotating speed; Adopt distilled water and dehydrated alcohol to intersect washing repeatedly, at 30 ℃ of following vacuum-drying 24h, obtain the Fe of oleic acid modification
3O
4Magnetic nano-particle;
Step 2: with the Fe of oleic acid modification
3O
4Magnetic nano-particle adds ultra-sonic dispersion in the normal hexane according to 0.5g/mL, forms the oil base magnetic fluid;
Step 3: be the lauryl sodium sulfate aqueous solution of 3.5mg/mL with concentration, join in the oil base magnetic fluid, form magnetic template emulsion, add methyl methacrylate MMA and methacrylic acid MAA more successively, mechanical stirring 2h under 60～70 ℃, 800r/min rotating speed, the Potassium Persulphate KPS, the Ethylene glycol dimethacrylate EGDMA of 0.4～0.6mL that continue to add 0.01g, thermal-initiated polymerization 20h obtains the Fe of carboxyl-functional
3O
4Magnetic nano-particle; The volume ratio of described methyl methacrylate MMA and methacrylic acid MAA is 2:1～4:1;
Step 4: thymopeptide-5 TP-5 and acrylamide AM are dissolved in the distilled water, behind 25 ℃ of following prepolymerization 20～60min, add the Fe of 1.2% carboxyl-functional of this distilled water quality
3O
4Magnetic nano-particle, the ultrasonic 15min of room temperature forms the water mixed solution; The mol ratio of described thymopeptide-5 TP-5 and acrylamide AM is 1:20～1:28;
Step 5: under the 800r/min stirring action, the potassium persulfate solution 1mL that adds EGDMA, the 1.5mg/mL of 0.3～0.5mL in the water mixed solution successively, in 30 ℃ of water-baths behind thermal-initiated polymerization 5～7h, be cooled to room temperature, adopt distilled water and dehydrated alcohol to intersect washing repeatedly, at 30 ℃ of following vacuum-drying 24h, obtain molecular blotting magnetic microsphere;
Step 6: molecular blotting magnetic microsphere is joined in the mixing solutions of phosphate buffered saline buffer and methyl alcohol, at 25 ℃, 120rmpmin
-1Shaking table vibration 10h is with wash-out thymopeptide-5 TP-5 under the rotating speed; Again with distilled water wash repeatedly, at 25 ℃ of following vacuum-drying 24h, obtain the thymopeptide-5 molecular blotting magnetic microsphere; The volume ratio of described phosphate buffered saline buffer and methyl alcohol is 60:40.
2. according to the preparation method of the described thymopeptide-5 molecular blotting magnetic microsphere of claim 1, it is characterized in that: the pH=7 of described phosphate buffered saline buffer.
3. according to the preparation method of the described thymopeptide-5 molecular blotting magnetic microsphere of claim 1, it is characterized in that: adopt distilled water and dehydrated alcohol to intersect in the step 1 and wash 6 times.
4. according to the preparation method of the described thymopeptide-5 molecular blotting magnetic microsphere of claim 1, it is characterized in that: step 5 adopts distilled water and dehydrated alcohol to intersect and washs 3 times.
5. according to the preparation method of the described thymopeptide-5 molecular blotting magnetic microsphere of claim 1, it is characterized in that: step 6 distilled water wash 6 times.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103816876A (en) * | 2014-02-25 | 2014-05-28 | 浙江大学宁波理工学院 | Preparation method of composite adsorption material capable of removing plasticizer in liquor |
| CN108376608A (en) * | 2018-02-10 | 2018-08-07 | 青岛大学 | A kind of magnetic nano-particle and its purposes for preparing Magnetic solid phases carrier |
| CN109021173A (en) * | 2018-06-21 | 2018-12-18 | 西北工业大学 | A kind of preparation method of thymopeptide-5 molecular engram core-shell particles |
| CN109810281A (en) * | 2018-12-27 | 2019-05-28 | 江苏大学 | It is a kind of based on polymethyl methacrylate nano ball-molecular engram fluorescent material preparation method and its usage |
| CN110711566A (en) * | 2019-09-30 | 2020-01-21 | 重庆东渝中能实业有限公司 | Molecular imprinting material of vitamin K1, and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080221248A1 (en) * | 2007-03-05 | 2008-09-11 | Sungkyunkwan University Foundation For Corporate Collaboration | Method for preparing surface-imprinted polymer microspheres in the form of core-shell for selective separation of heavy metal ions |
| CN102585119A (en) * | 2012-02-23 | 2012-07-18 | 宁波市疾病预防控制中心 | Preparation method of magnetic nanometer molecular imprinting composite material related to estrogen |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080221248A1 (en) * | 2007-03-05 | 2008-09-11 | Sungkyunkwan University Foundation For Corporate Collaboration | Method for preparing surface-imprinted polymer microspheres in the form of core-shell for selective separation of heavy metal ions |
| CN102585119A (en) * | 2012-02-23 | 2012-07-18 | 宁波市疾病预防控制中心 | Preparation method of magnetic nanometer molecular imprinting composite material related to estrogen |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103816876A (en) * | 2014-02-25 | 2014-05-28 | 浙江大学宁波理工学院 | Preparation method of composite adsorption material capable of removing plasticizer in liquor |
| CN108376608A (en) * | 2018-02-10 | 2018-08-07 | 青岛大学 | A kind of magnetic nano-particle and its purposes for preparing Magnetic solid phases carrier |
| CN109021173A (en) * | 2018-06-21 | 2018-12-18 | 西北工业大学 | A kind of preparation method of thymopeptide-5 molecular engram core-shell particles |
| CN109810281A (en) * | 2018-12-27 | 2019-05-28 | 江苏大学 | It is a kind of based on polymethyl methacrylate nano ball-molecular engram fluorescent material preparation method and its usage |
| CN109810281B (en) * | 2018-12-27 | 2021-08-03 | 江苏大学 | Preparation method and application of polymethyl methacrylate nanosphere-based molecularly imprinted fluorescent material |
| CN110711566A (en) * | 2019-09-30 | 2020-01-21 | 重庆东渝中能实业有限公司 | Molecular imprinting material of vitamin K1, and preparation method and application thereof |
| CN110711566B (en) * | 2019-09-30 | 2022-06-17 | 重庆东渝中能实业有限公司 | Molecular imprinting material of vitamin K1, and preparation method and application thereof |
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