CN113813935A - Temperature response type protein imprinting magnetic nano-microsphere and preparation method thereof - Google Patents
Temperature response type protein imprinting magnetic nano-microsphere and preparation method thereof Download PDFInfo
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- CN113813935A CN113813935A CN202111118440.3A CN202111118440A CN113813935A CN 113813935 A CN113813935 A CN 113813935A CN 202111118440 A CN202111118440 A CN 202111118440A CN 113813935 A CN113813935 A CN 113813935A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
- B01J20/268—Polymers created by use of a template, e.g. molecularly imprinted polymers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
- B01J20/28002—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their physical properties
- B01J20/28009—Magnetic properties
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
- B01J20/28014—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their form
- B01J20/28016—Particle form
- B01J20/28021—Hollow particles, e.g. hollow spheres, microspheres or cenospheres
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/305—Addition of material, later completely removed, e.g. as result of heat treatment, leaching or washing, e.g. for forming pores
- B01J20/3057—Use of a templating or imprinting material ; filling pores of a substrate or matrix followed by the removal of the substrate or matrix
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/76—Albumins
- C07K14/765—Serum albumin, e.g. HSA
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/795—Porphyrin- or corrin-ring-containing peptides
- C07K14/805—Haemoglobins; Myoglobins
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2462—Lysozyme (3.2.1.17)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01017—Lysozyme (3.2.1.17)
Abstract
The invention provides a temperature response type protein imprinting magnetic nano-microsphere and a preparation method thereof, belonging to the technical field of protein separation engineering. The preparation method of the temperature response type protein imprinting magnetic nano microsphere comprises the following steps: firstly, synthesizing a magnetic nano material by a coprecipitation method, then wrapping a silicon dioxide protective layer on the surface of a magnetic microsphere by an improved stober method, and further introducing halogen atoms on the surface of the protective layer; the halogen atom initiates the polymerization of the temperature sensitive functional monomer under the action of a ligand-copper catalytic system to obtain the protein imprinted magnetic nano-microsphere. The microsphere has uniform size and good stability, can control the adsorption and desorption of protein by changing the environmental temperature, has specific recognition capability on template protein, has the maximum adsorption capacity of 268mg/g, has adsorption equilibrium time of 5min, and can realize the rapid separation of protein under the action of a magnetic field. The preparation condition is mild, the equipment is simple, and the desorbed molecularly imprinted nanospheres can be recycled. Provides a rapid and efficient means for separating and purifying target protein in complex biological samples. Has good application prospect in the fields of separation and purification of biomacromolecules, medicine and other fields.
Description
Technical Field
The invention relates to the technical field of protein separation engineering, in particular to a temperature response type protein imprinting magnetic nano-microsphere and a preparation method thereof.
Background
Proteins are important substances in life activities and also important raw materials in the biomedical field. With the development of medical technology, the development of highly selective and efficient analytical methods has been the focus of attention, since proteins can provide solutions to most diseases. Thus, the market for protein separation materials continues to grow.
At present, high performance liquid chromatography, capillary electrophoresis and electrochromatography show great potential in the separation of complex biological samples. But the equipment is expensive and the operation is complex. Affinity chromatography is generally considered to be one of the most efficient methods for specifically capturing target proteins. However, the method has the disadvantages of easy inactivation, high cost, difficult antibody acquisition and the like, and the application of the technology is hindered. The affinity chromatographic column is prepared by a chemical synthesis method, the raw materials are cheap and easy to obtain, the quality consistency is good, the product performance is stable, and the defects can be overcome.
The molecular imprinting technology is a preparation technology which is integrally inosculated with a target molecule (template molecule) in space and a binding site, and the prepared polymer is called as a molecular imprinting polymer and has larger adsorption capacity and higher selectivity to the target molecule. However, when used for imprinting of larger protein molecules, the diffusion of the protein molecules in the polymer is slow or even impossible; proteins are only soluble in water and are labile to inactivation, and organic solvents commonly used in small molecular imprinting cannot be used. The surface molecular imprinting method is characterized in that a specific material is used as a carrier, in the preparation process, a functional monomer is combined with a template molecule at an emulsion interface, and after a cross-linking agent and the functional monomer are polymerized, the structure is fixed on the surface of a matrix or an imprinted polymer, so that imprinting with a certain recognition effect is generated. Compared with the traditional bulk polymerization method, the imprinted polymer prepared by the method has imprinted sites positioned on or close to the surface of the molecularly imprinted material, and the elution and recombination rates of the template molecules are higher; the preparation process can be carried out in aqueous solution, and the preparation method is simple to operate and high in stability. Especially for biological macromolecules with larger steric hindrance, the advantages of surface molecular imprinting are more obvious. In view of the above advantages, reports of the preparation of a polymer having a large molecular imprinting area by a surface imprinting method have been increasing in recent years.
The Jiaxianu takes bovine hemoglobin (BHb) as a template molecule and dopamine as a functional monomer and a cross-linking agent to silanize Fe3O4The particles are used as reaction carriers, and bovine hemoglobin magnetic molecularly imprinted polymer (BHb-MIP) is synthesized by a surface imprinting method. The adsorption experiment results show that: when the concentration of the BHb solution is 0.2 mg/mL, the maximum adsorption capacity (4.65 +/-0.38 mg/g) is reached at 30min, and the imprinted polymer has higher selectivity (beta =2.19) and stability to the template protein. After removal of the template protein with the eluent SDS-acetic acid (0.1% w/v: 3% v/v), the imprinted polymer can be reused more than three times. (Jiaxianping. protein blotting technical research and application based on magnetic nano particles and temperature-sensitive hydrogel [ D)]University of Hunan.)
Double bond functionalized Fe of Guomilitary nepheline3O4The magnetic molecularly imprinted polymer (MIP-Fe) of the bovine hemoglobin is synthesized by the surface imprinting technology by taking the nano particles as a carrier, the bovine hemoglobin (BHb) as a template molecule, methacrylic acid (MAA) and Itaconic Acid (IA) as bifunctional monomers and N, N-Methylene Bisacrylamide (MBA) as a cross-linking agent3O4@ AA). The thermodynamic adsorption equilibrium concentration is 1.0 mg/mL, the theoretical maximum adsorption capacity is 117.27mg/mL, the imprinting factor is 4.43, and the good adsorption performance and specific selectivity performance are shown for the template protein. The template molecules can be recycled for at least 5 times after being eluted by shaking with 2% (w/v) SDS-2% (v/v) HAc solution. (Guoqianxian. preparation and identification performance research of protein imprinted polymer on surface of magnetic nano material carrier [ D]University of Hunan, 2016)
Qiubuixia firstly uses chemical precipitation method to prepare Fe3O4Nanoparticles, rear pair Fe3O4The nano particles are modified by O-Si-O, -COOH and other groups to obtain Fe3O4@SiO2-AA particles followed by the selection of methylpropanePreparing Fe by using olefine acid (MAA) and Itaconic Acid (IA) as functional monomers and N, N-Methylene Bisacrylamide (MBA) as a cross-linking agent through a molecular imprinting technology3O4@SiO2-AA-MIP particles. Adsorption kinetics shows that the adsorption of MIP particles to BHb is balanced when the MIP particles are adsorbed for 12 hours, the maximum adsorption amount of 116.69 mg/g particles is obtained, competitive adsorption experiments show that the MIP particles have specific selectivity to target protein, and 1% (w/v) SDS solution has the best elution effect on BHb molecules on the MIP particles. (Qiubuixia. magnetic particle composite molecular imprinting for protein separation and purification research [ D)]Jiangnan university.)
The above documents show that the conventional molecularly imprinted polymers require a large amount of salt, acid and alkali solutions to achieve elution of target molecules. Harsh elution conditions are easy to cause denaturation and inactivation of target protein, so that the protein loses use value. The use of a large amount of eluent does not accord with the development concept of green chemistry, the subsequent treatment cost is increased, and certain pressure is also caused to the environmental protection.
The critical transition temperature of the N-isopropylacrylamide is 32 ℃, the temperature is close to the physiological temperature of a human body, and when the external temperature is higher than the temperature, the polymer is in a shrinkage state; when the temperature is lower than this temperature, it assumes a swollen state. Therefore, the polymer state can be regulated and controlled by adjusting the external temperature, so that the capture and release of the template molecules are realized, and the polymer is a mainstream material of a protein molecular imprinting framework.
Chinese patent CNIO2532408A discloses a preparation method of temperature-sensitive magnetic western-blotting nanospheres, which uses bovine serum protein as template molecules, N-isopropyl acrylamide, acrylamide and N- (3-dimethylaminopropyl) methacrylamide as functional monomers, silicon-coated magnetic nanospheres as carriers, N-methylene bisacrylamide as a cross-linking agent, and Ammonium Persulfate (APS) and Tetramethylethylenediamine (TEMED) redox initiation system to polymerize and prepare the molecularly-blotting nanospheres. The molecular imprinting nanosphere has good selectivity on the template protein, and realizes the adsorption and desorption on the template protein through the change of temperature. However, the polymerization process has uncontrollable irregular imprinted sites and is easy to crosslink into large particles, which hinders the diffusion of the template protein and leads to the reduction of the adsorption capacity. Therefore, the development of the protein imprinting material with the specific recognition function, which has large adsorption capacity, can be quickly separated and is easy to desorb, has important practical application value.
Disclosure of Invention
Aiming at the problems, the invention provides a temperature response type protein imprinting magnetic nano microsphere and a preparation method thereof, and Fe is synthesized by a coprecipitation method3O4Then, a silicon dioxide protective layer is wrapped on the surface of the magnetic microsphere by adopting an improved stober method, halogen atoms are introduced on the surface of the protective layer, and finally, the polymerization reaction of the imprinted polymer is initiated on the surface of the magnetic microsphere at normal temperature. The obtained polymer has a definite topological structure and chemical composition, imprinting sites are uniformly distributed on the surface of the microsphere, and the polymer has a specific recognition function on template protein, is large in adsorption capacity, can be quickly separated and is easy to desorb. The preparation raw materials of the material are easy to obtain, the process is simple, the reaction condition is mild, the cost is low, and the quality consistency is good. In order to achieve the above object, the present invention adopts the following technical solutions: adding the magnetic microspheres introduced with halogen atoms, a certain amount of N-isopropylacrylamide, an auxiliary monomer, a crosslinking agent, template molecules and 10-100 mL of phosphoric acid buffer solution into a three-neck flask reactor, performing ultrasonic dispersion and self-assembly at 4-37 ℃ for 30-120 min, bubbling nitrogen for 20-180 min to remove oxygen in the system, finally adding a catalyst in a nitrogen atmosphere, and mechanically stirring and polymerizing at room temperature for 6-24 h. And collecting the synthesized temperature response type protein imprinting magnetic nano-microspheres by using an external magnetic field. Washing the product with deionized water for several times to remove unreacted monomers and impurities; and removing the template molecules by using eluent to obtain the molecularly imprinted nanospheres.
The invention has the advantages of
(1) The invention has the advantages of cheap and easily obtained raw materials, mild preparation conditions, simple equipment and simple process.
(2) The invention takes the initiator as the imprinting carrier, the imprinting sites are uniformly distributed on the surface of the microsphere, and the obtained polymer has a definite topological structure and chemical composition. Solves the contradiction that the protein molecular imprinting technology requires high specific surface area, large molecular diffusion coefficient and easy separation.
(3) The microsphere prepared by the invention is a core-shell structure taking ferroferric oxide nano microsphere as a core and temperature response copolymer as a shell layer. The shell layer contains complementary functional groups of amino acid residues such as N-isopropyl acrylamide, tert-butyl acrylamide, acrylic acid and the like, and a three-dimensional structure matched with the volume of a target molecule is constructed through template molecules. With the change of the ambient temperature, the molecularly imprinted polymer shows a reversible change of a shrinkage-swelling state. The method has the advantages that the adsorption and desorption of the protein can be controlled only by controlling the temperature change, and the use of salt, acid and alkali solution in the traditional separation method is avoided; the separation of the molecularly imprinted polymer from the protein can be realized through magnetic response.
(4) The microspheres prepared by the method have uniform size and good stability, have specific recognition capability on template protein, have the maximum adsorption capacity of 268mg/g, have adsorption equilibrium time of 5min, and can realize rapid separation of protein under the action of a magnetic field. The performance is superior to that of the protein imprinted polymer reported in the prior literature. Provides a rapid and efficient means for separating and purifying target protein in complex biological samples. Has good application prospect in the fields of separation and purification of biomacromolecules, medicine and other fields.
Drawings
FIG. 1 is a scanning electron microscope photograph of initiator functionalized nanospheres prepared in example 1; FIG. 2 is a scanning electron micrograph of the temperature-responsive Western-imprinted magnetic nanospheres prepared in example 1. The scanning electron microscope images 1 and 2 show that the molecular imprinting nanospheres prepared by the method have uniform size and good monodispersity. FIG. 3 is a FT-IR spectrum of the temperature-responsive Western-imprinted magnetic nanospheres prepared in example 1; FIG. 4 is the temperature response protein print magnetic nanometer microsphere prepared in example 11HNMR spectrogram. By FT-IR,1The molecular imprinting nanosphere structure prepared by the method is characterized by HNMR means, and the results of figures 3 and 4 show that the temperature response protein imprinting magnetic nanospheres are successfully prepared by the method. FIG. 5 is a temperature-responsive protein stamp prepared in example 1SDS-PAGE analysis chart of LYZ selective adsorption in egg white by the trace magnetic nano microspheres. Lane 1: a standard molecular weight protein band; lane 2: egg white protein strips; lane 3: egg white protein strips after molecular imprinting nanospheres are adsorbed; lane 4: desorbing the LYZ strip; SDS-PAGE analysis shows that the nano microsphere prepared by the method has specific adsorption capacity on LYZ and can achieve the aim of desorption by changing the temperature. FIG. 6 is a graph of the kinetic adsorption UV-Vis of the temperature response protein imprinted magnetic nanospheres prepared in example 1. As shown in FIG. 6, the molecularly imprinted nanospheres prepared by the method have a large adsorption capacity for LYZ, and can reach adsorption equilibrium within 5 min. FIG. 7 is a graph of the magnetic response separation effect of the temperature response Western blotting magnetic nanospheres prepared in example 1.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without any inventive step, are within the scope of the present invention.
Example 1
Dissolving NIPAM 0.747g, MBA 22.0mg, MAA 30.0 μ L, and LYZ 0.15g in 20.0ml PBS buffer solution (concentration 35mmoL/L, pH 7.4), adding Fe 1.00g3O4@SiO2Ultrasonically dispersing and self-assembling Br at 25 ℃ for 30min, bubbling nitrogen for 60min to remove oxygen in the reaction system, then adding 50.0mg of CuBr and 78.0 mu of LPMDETA into a nitrogen atmosphere, mechanically stirring and polymerizing at room temperature for 24h, then externally applying a magnetic field for separation, and carrying out self-assembly on the mixture by using acetic acid (2%, V/V) and SDS (2%, W/V) 1: 1, washing the mixed solution for many times to remove template molecules, and drying in vacuum at 60 ℃ to obtain the molecular imprinting nano-microspheres. The electron microscope test result shows that the particle size is uniform and monodisperse, and the average particle size is 120 nm. The microspheres are used for lysozyme adsorption experiments, the adsorption equilibrium time is 5min, and the maximum adsorption capacity reaches 268 mg/g. The nano-microsphere is used for lysozyme adsorption in egg white, and SDS-PAGE analysis results show that the nano-microsphere has specific selective adsorption capacity for lysozyme.
Example 2
Dissolving 0.747g NIPAM, 28.14mg EGDMA, 30.0 μ L MAA, and 0.16g LYZ in 20.0ml PBS buffer solution (concentration 30mmoL/L, pH 7.4), adding 1.00g Fe3O4@SiO2Ultrasonically dispersing and self-assembling Br at 25 ℃ for 40min, bubbling nitrogen for 80min to remove oxygen in the reaction system, then adding 50.0mg of CuBr and 78.0 mu L of PMDETA into nitrogen atmosphere, mechanically stirring and polymerizing for 20h at room temperature, then externally adding a magnetic field for separation, and carrying out self-assembly by using acetic acid (2%, V/V) and SDS (2%, W/V) 1: 1, washing the mixed solution for many times to remove template molecules, and drying in vacuum at 60 ℃ to obtain the molecular imprinting nano-microspheres. The electron microscope test result shows that the particle size is uniform and monodisperse, and the average particle size is 130 nm. The microspheres are used for lysozyme adsorption experiments, the adsorption equilibrium time is 5min, and the maximum adsorption capacity reaches 260 mg/g. The microspheres are used for lysozyme adsorption in egg white, and SDS-PAGE analysis results show that the nano microspheres have specific selective adsorption capacity on lysozyme.
Example 3
0.680g of NIPAM, 20.0mg of MBA, 30.0mg of AM, and 20.0mg of BSA were dissolved in 20.0ml of PBS buffer solution (concentration: 35mmoL/L, pH: 6.4), and 0.40g of Fe was added3O4@SiO2Ultrasonically dispersing and self-assembling Br at 25 ℃ for 40min, bubbling nitrogen for 60min to remove oxygen in the reaction system, then adding 50.0mg of CuBr and 78.0 mu L of PMDETA into nitrogen atmosphere, mechanically stirring and polymerizing for 24h at room temperature, then externally adding a magnetic field for separation, and carrying out self-assembly by using acetic acid (2%, V/V) and SDS (2%, W/V) 1: 1, washing the mixed solution for many times to remove template molecules, and drying in vacuum at 60 ℃ to obtain the molecular imprinting nano-microspheres. The electron microscope test result shows that the particle size is uniform and monodisperse, and the average particle size is 120 nm. The microspheres are used for bovine serum albumin adsorption experiments, the adsorption balance time is 5min, and the maximum adsorption capacity reaches 286 mg/g. The microsphere is used for competitive adsorption of mixed protein of lysozyme, bovine serum albumin and bovine hemoglobin, and SDS-PAGE analysis results show that the nano microsphere has specific selective adsorption capacity on bovine serum albumin.
Example 4
Dissolving NIPAM 0.70g, MBA 22.0mg, AM 30.0mg, MAA 25.0 μ L, and LYZ 0.20g in 20.0 mLP.0To a buffer solution of BS (30 mmoL/L concentration, pH 6.8), 1.00g of Fe was added3O4@SiO2Ultrasonically dispersing and self-assembling Br at 25 ℃ for 50min, bubbling nitrogen for 80min to remove oxygen in the reaction system, then adding 50.0mg of CuBr and 78.0 mu L of PMDETA into nitrogen atmosphere, mechanically stirring and polymerizing for 6h at room temperature, then externally adding a magnetic field for separation, and carrying out self-assembly by using acetic acid (2%, V/V) and SDS (2%, W/V) 1: 1, washing the mixed solution for many times to remove template molecules, and drying in vacuum at 60 ℃ to obtain the molecular imprinting nano-microspheres. The microspheres are used for lysozyme adsorption experiments, the adsorption equilibrium time is 5min, and the maximum adsorption capacity reaches 268 mg/g. The microspheres are used for lysozyme adsorption in egg white, and SDS-PAGE analysis results show that the nano microspheres have specific selective adsorption capacity on lysozyme.
Example 5
Dissolving 0.68g NIPAM, 22.0mg MBA, 10.0mg tBA, 30.0 μ L AAc, 0.15g LYZ in 20.0ml PBS buffer solution (concentration 35mmoL/L, pH 7.4), adding 1.00g Fe3O4@SiO2Ultrasonically dispersing and self-assembling Br at 25 ℃ for 30min, bubbling nitrogen for 60min to remove oxygen in the reaction system, then adding 50.0mg of CuBr and 78.0 mu L of PMDETA into nitrogen atmosphere, mechanically stirring and polymerizing for 12h at room temperature, then externally adding a magnetic field for separation, and carrying out self-assembly by using acetic acid (2%, V/V) and SDS (2%, W/V) 1: 1, washing the mixed solution for many times to remove template molecules, and drying in vacuum at 60 ℃ to obtain the molecular imprinting nano-microspheres. The microspheres are used for lysozyme adsorption experiments, the adsorption equilibrium time is 5min, and the maximum adsorption capacity reaches 268 mg/g. The microspheres are used for lysozyme adsorption in egg white, and SDS-PAGE analysis results show that the nano microspheres have specific selective adsorption capacity on lysozyme.
Example 6
0.70g of NIPAM, 10.0mg of 1, 4-butanediol dimethacrylate, 30.0. mu.L of MAA and 0.30g of BSA were dissolved in 20.0ml of a buffer solution (concentration: 35mmoL/L, pH 7.4) of PBS, and 1.00g of Fe was added3O4@SiO2Ultrasonically dispersing and self-assembling-Br at 25 deg.C for 60min, bubbling nitrogen for 90min to remove oxygen in the reaction system, adding 50.0mg CuBr and 78.0 μ L PMDETA in nitrogen atmosphere, mechanically stirring at room temperature for 20 hr, externally adding magnetic field for separation, and treating with acetic acid at low temperature(2%, V/V) and SDS (2%, W/V) 1: 1, washing the mixed solution for many times to remove template molecules, and drying in vacuum at 60 ℃ to obtain the molecular imprinting nano-microspheres. The electron microscope test result shows that the particle size is uniform and monodisperse, and the average particle size is 125 nm. The microspheres are used for bovine serum albumin adsorption experiments, the adsorption balance time is 5min, and the maximum adsorption capacity reaches 286 mg/g. The microsphere is used for competitive adsorption of mixed protein of lysozyme, bovine serum albumin and bovine hemoglobin, and SDS-PAGE analysis results show that the nano microsphere has specific selective adsorption capacity on bovine serum albumin.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (7)
1. A preparation method of temperature response type protein imprinting magnetic nano-microspheres comprises the following steps: firstly, synthesizing a magnetic nano material by a coprecipitation method, then wrapping a silicon dioxide protective layer on the surface of a magnetic microsphere by an improved stober method, and further introducing halogen atoms on the surface of the protective layer; and then adding the magnetic microspheres introduced with halogen atoms, a certain amount of N-isopropylacrylamide, an auxiliary monomer, a crosslinking agent, template molecules and 10-100 mL of phosphoric acid buffer solution into a three-neck flask reactor, performing ultrasonic dispersion self-assembly at 4-37 ℃ for 30-120 min, bubbling nitrogen for 20-180 min to remove oxygen in the system, finally adding a catalyst in a nitrogen atmosphere, and performing mechanical stirring polymerization at room temperature for 6-24 h.
2. And collecting the synthesized temperature response type protein imprinting magnetic nano-microspheres by using an external magnetic field.
3. Washing the product with deionized water for several times to remove unreacted monomers and impurities; and removing the template molecules by using eluent to obtain the protein imprinted nano-microspheres.
4. The temperature-responsive western-imprinted magnetic nanosphere according to claim 1, wherein: the microsphere takes ferroferric oxide nano microsphere as a core and takes a temperature response copolymer as a shell layer.
5. The shell layer contains complementary functional groups of amino acid residues such as N-isopropyl acrylamide, tert-butyl acrylamide, acrylic acid and the like, and has a three-dimensional structure matched with the volume of a target molecule.
6. The adsorption and desorption of protein molecules are realized along with the change of the external environment temperature; the separation of the molecularly imprinted polymer from the protein is realized by the change of the magnetic field.
7. The preparation method of the temperature-responsive western-imprinted magnetic nanosphere according to claim 1, wherein the preparation method comprises the following steps: the mol ratio of the N-isopropylacrylamide, the auxiliary monomer and the cross-linking agent is 94-75: 5-20: 1-5, and the amount of the template protein is 5-40% of the mass of the functional monomer;
the preparation method of the temperature-responsive western-imprinted magnetic nanosphere according to claim 1, wherein the preparation method comprises the following steps: the auxiliary monomer is one or a mixture of more of methacrylic acid, N-tertiary butyl acrylamide, acrylic acid and acrylamide;
the preparation method of the temperature-responsive western-imprinted magnetic nanosphere according to claim 1, wherein the preparation method comprises the following steps: the cross-linking agent is one or a mixture of more of N, N' -methylene bisacrylamide, ethylene glycol methacrylate, triethylene glycol dimethacrylate, 1, 3-butanediol dimethacrylate, 1, 4-butanediol dimethacrylate, 1, 6-hexanediol dimethacrylate and bisphenol A dimethacrylate;
the preparation method of the temperature-responsive western-imprinted magnetic nanosphere according to claim 1, wherein the preparation method comprises the following steps: preparing Fe3O4 particles by adopting a coprecipitation method, then coating a silicon dioxide thin layer on the surface of a seed solution by taking Fe3O4 particles as seeds, wherein a magnetic core coated by SiO2 has monodisperse superparamagnetism, and the particle size is 80-300 nm;
the preparation method of the temperature-responsive western-imprinted magnetic nanosphere according to claim 1, wherein the preparation method comprises the following steps: the magnetic core is functionalized with 2-bromoisobutyryl bromide;
the preparation method of the temperature-responsive western-imprinted magnetic nanosphere according to claim 1, wherein the preparation method comprises the following steps: the polymerization reaction of the molecularly imprinted polymer is initiated by the surface of the magnetic particle;
the preparation method of the temperature-responsive western-imprinted magnetic nanosphere according to claim 1, wherein the preparation method comprises the following steps: the buffer solution is a dipotassium hydrogen phosphate system and a potassium dihydrogen phosphate system, the concentration is 10-100 mmoL/L, and the pH value is 6-8;
the preparation method of the temperature-responsive western-imprinted magnetic nanosphere according to claim 1, wherein the preparation method comprises the following steps: the prepared molecularly imprinted nanospheres are applied to protein enrichment, separation and purification.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101347721A (en) * | 2008-09-17 | 2009-01-21 | 南开大学 | Method for preparing protein magnetic blotting nanospheres |
| CN101816927A (en) * | 2010-04-30 | 2010-09-01 | 南开大学 | Temperature-sensitive protein molecular engram monolithic column and preparation method and application thereof |
| CN102532408A (en) * | 2011-12-27 | 2012-07-04 | 南开大学 | Preparation method of temperature sensitive magnetic western-blotting nanosphere |
| US20120270964A1 (en) * | 2009-12-01 | 2012-10-25 | Cranfield University | Preparation of molecularly imprinted polymers |
| CN103100377A (en) * | 2013-02-27 | 2013-05-15 | 江苏大学 | Green preparation method of magnetic molecular imprinting sorbent |
| CN104844768A (en) * | 2015-04-22 | 2015-08-19 | 中南大学 | Preparation method and application of core-shell structure temperature sensitive magnetic protein imprinted microspheres |
| CN105017476A (en) * | 2015-07-14 | 2015-11-04 | 辽宁师范大学 | Preparation method for hemoglobin molecularly imprinted polymers through autocatalysis of ATRP |
| CN110189882A (en) * | 2019-06-24 | 2019-08-30 | 陕西科技大学 | A kind of carbon dioxide gas stimuli responsive magnetic Nano material and preparation method thereof with western blot |
-
2021
- 2021-09-24 CN CN202111118440.3A patent/CN113813935A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101347721A (en) * | 2008-09-17 | 2009-01-21 | 南开大学 | Method for preparing protein magnetic blotting nanospheres |
| US20120270964A1 (en) * | 2009-12-01 | 2012-10-25 | Cranfield University | Preparation of molecularly imprinted polymers |
| CN101816927A (en) * | 2010-04-30 | 2010-09-01 | 南开大学 | Temperature-sensitive protein molecular engram monolithic column and preparation method and application thereof |
| CN102532408A (en) * | 2011-12-27 | 2012-07-04 | 南开大学 | Preparation method of temperature sensitive magnetic western-blotting nanosphere |
| CN103100377A (en) * | 2013-02-27 | 2013-05-15 | 江苏大学 | Green preparation method of magnetic molecular imprinting sorbent |
| CN104844768A (en) * | 2015-04-22 | 2015-08-19 | 中南大学 | Preparation method and application of core-shell structure temperature sensitive magnetic protein imprinted microspheres |
| CN105017476A (en) * | 2015-07-14 | 2015-11-04 | 辽宁师范大学 | Preparation method for hemoglobin molecularly imprinted polymers through autocatalysis of ATRP |
| CN110189882A (en) * | 2019-06-24 | 2019-08-30 | 陕西科技大学 | A kind of carbon dioxide gas stimuli responsive magnetic Nano material and preparation method thereof with western blot |
Non-Patent Citations (2)
| Title |
|---|
| JINGJING ZHOU等: ""Surface molecularly imprinted thermo-sensitive polymers based on lightweight hollow magnetic microspheres for specific recognition of BSA"", 《APPLIED SURFACE SCIENCE》, vol. 486, pages 265 - 273 * |
| 盖青青等: ""牛血清白蛋白和溶菌酶为双模板蛋白质的表面印迹聚合物的制备及吸附性能"", 《色谱》, vol. 33, no. 5, pages 475 - 480 * |
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