CN111533865B - Anti-protein-adsorption self-cleaning block copolymer, preparation method and application thereof - Google Patents

Anti-protein-adsorption self-cleaning block copolymer, preparation method and application thereof Download PDF

Info

Publication number
CN111533865B
CN111533865B CN202010432143.5A CN202010432143A CN111533865B CN 111533865 B CN111533865 B CN 111533865B CN 202010432143 A CN202010432143 A CN 202010432143A CN 111533865 B CN111533865 B CN 111533865B
Authority
CN
China
Prior art keywords
peo
block copolymer
capillary
reaction
self
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202010432143.5A
Other languages
Chinese (zh)
Other versions
CN111533865A (en
Inventor
于冰
丛海林
王芳
申有青
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Qingdao University
Original Assignee
Qingdao University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Qingdao University filed Critical Qingdao University
Priority to CN202010432143.5A priority Critical patent/CN111533865B/en
Publication of CN111533865A publication Critical patent/CN111533865A/en
Application granted granted Critical
Publication of CN111533865B publication Critical patent/CN111533865B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F293/00—Macromolecular compounds obtained by polymerisation on to a macromolecule having groups capable of inducing the formation of new polymer chains bound exclusively at one or both ends of the starting macromolecule
    • C08F293/005—Macromolecular compounds obtained by polymerisation on to a macromolecule having groups capable of inducing the formation of new polymer chains bound exclusively at one or both ends of the starting macromolecule using free radical "living" or "controlled" polymerisation, e.g. using a complexing agent
    • 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
    • C07K1/24—Extraction; Separation; Purification by electrochemical means
    • C07K1/26—Electrophoresis
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F220/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical or a salt, anhydride ester, amide, imide or nitrile thereof
    • C08F220/02—Monocarboxylic acids having less than ten carbon atoms; Derivatives thereof
    • C08F220/10—Esters
    • C08F220/38—Esters containing sulfur
    • C08F220/382—Esters containing sulfur and containing oxygen, e.g. 2-sulfoethyl (meth)acrylate
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F220/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical or a salt, anhydride ester, amide, imide or nitrile thereof
    • C08F220/02—Monocarboxylic acids having less than ten carbon atoms; Derivatives thereof
    • C08F220/52—Amides or imides
    • C08F220/54—Amides, e.g. N,N-dimethylacrylamide or N-isopropylacrylamide
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
    • C09D153/00—Coating compositions based on block copolymers containing at least one sequence of a polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds; Coating compositions based on derivatives of such polymers
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
    • C09D5/00—Coating compositions, e.g. paints, varnishes or lacquers, characterised by their physical nature or the effects produced; Filling pastes
    • C09D5/16—Antifouling paints; Underwater paints
    • C09D5/1656—Antifouling paints; Underwater paints characterised by the film-forming substance
    • C09D5/1662—Synthetic film-forming substance
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
    • C08F2438/00—Living radical polymerisation
    • C08F2438/01—Atom Transfer Radical Polymerization [ATRP] or reverse ATRP
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P20/00—Technologies relating to chemical industry
    • Y02P20/50—Improvements relating to the production of bulk chemicals
    • Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids

Abstract

The invention discloses an anti-protein adsorption self-cleaning block copolymer, a preparation method and application thereof, wherein the block copolymer is prepared by taking three hydrophilic materials of polyethylene glycol monomethyl ether (PEO), N-isopropyl acrylamide (NIPAAm) and 3-sulfopropyl methacrylate (SPMAP) as raw materials and carrying out Atom Transfer Radical Polymerization (ATRP) on a macroinitiator. Each block in the triblock copolymer PEO-PNIPAAM-PSPMAP has good hydrophilicity, so that the protein adsorption resistance is guaranteed, and the temperature-sensitive block part provides a foundation for the self-cleaning performance. The copolymer and the coupling agent are crosslinked in a water phase in a layer-by-layer self-assembly mode to form a coating capillary with double effects of protein adsorption resistance and self-cleaning, and the application prospect is wide.

Description

Anti-protein-adsorption self-cleaning block copolymer, preparation method and application thereof
Technical Field
The invention relates to the technical field of antifouling coatings, in particular to an anti-protein adsorption self-cleaning block copolymer, a preparation method and application thereof.
Background
The protein is a bioactive macromolecule composed of amino acids, and the non-selective adsorption of the protein in a biological environment on the surface of a material can cause non-directional adsorption, which is a main cause for the surface pollution of the material. The materials should avoid as much as possible the induction of immune reactions in vivo in biomedical applications, and therefore have higher requirements. However, protein adsorption problems in the field of chemical analysis also affect the sensitivity of device detection and the efficiency of analysis.
Capillary electrophoresis has the characteristics of high sensitivity, high resolution, less sample introduction, low solvent consumption, high speed, various detection methods and the like, and is rapidly developed and widely applied in the fields of medicine detection, life science, environmental protection, food inspection and the like, particularly in the aspect of separation and analysis of various biological macromolecules such as protein, RNA and the like. However, the inner wall surface of the unmodified capillary generates nonspecific adsorption on the inner wall of the capillary due to the influence of electrostatic interaction, van der waals force, hydrogen bond interaction and the like with proteins, thereby influencing the separation and analysis of the proteins in a complex system.
The coating modification of the inner wall of the capillary is the most effective and common method for improving the separation effect and reproducibility of capillary electrophoresis and inhibiting the adsorption between an analyte and the inner wall of the capillary, so that a novel self-cleaning double-effect material with ultralow protein pollution behavior is designed and applied to the capillary coating, the optimization of protein separation and analysis is facilitated, and the cyclic regeneration and use of the coated column can be realized.
Disclosure of Invention
The invention aims to provide a block copolymer with double effects of protein adsorption resistance and self-cleaning, which can be used as a coating material for capillary electrophoresis and solves the problems of difficult protein adsorption and cleaning of the existing capillary wall coating material.
In order to solve the technical problems, the invention adopts the following technical scheme:
the block copolymer is a triblock copolymer PEO-PNIPAAm-PSPMAP prepared by using three hydrophilic materials of polyethylene glycol monomethyl ether (PEO), N-isopropyl acrylamide (NIPAAm) and methacrylic acid 3-sulfopropyl ester (SPMAP) as raw materials through an Atom Transfer Radical Polymerization (ATRP), and the structural formula of the triblock copolymer is shown as a formula (I):
Figure BDA0002500969850000021
wherein PEO is an initiation block, NIPAAm is a temperature sensitive block, and SPMAP is a cross-linking block;
in the formula (I), m is an integer of 16-35, and n is an integer of 8-15.
A preparation method of an anti-protein adsorption self-cleaning block copolymer comprises the following steps:
s1, under the protection of nitrogen atmosphere, adding 2-bromoisobutyryl bromide (BIBB), triethylamine (ETA) and dimethyl pyridine (DMAP) into a proper amount of organic solvent A, stirring to dissolve, and placing in an ice water bath to obtain a mixed solution B; dissolving polyethylene glycol monomethyl ether (PEO) in a proper amount of organic solvent A, mixing to obtain a mixed solution C, then dropwise adding the mixed solution C into the mixed solution B, reacting at room temperature for 10-20 h, extracting with NaCl solution for three times, drying an organic phase with anhydrous magnesium sulfate, filtering, removing the solvent by rotary evaporation, settling a product in glacial ethyl ether, filtering, and drying in vacuum to obtain a light yellow macromolecular initiator (PEO-Br);
s2, adding the initiators PEO-Br, NIPAAm and PMDETA prepared in the step S1 into a reaction vessel, adding a mixed solvent D for dissolving, vacuumizing and introducing nitrogen for three times, adding a catalyst under the protection of nitrogen, continuously introducing nitrogen for 10-20 min for sealing, placing the reaction vessel into an oil bath at 30 ℃, stirring for reacting for 3-5 h, and exposing the reaction vessel to air after the reaction is finished so as to terminate the reaction; THF is used as an eluent, the reaction mixture is subjected to alkaline alumina column, a large amount of solvent is removed by rotary evaporation, the product is dropped into ethyl glacial ether for sedimentation and washing for 3 times, and the product is filtered and dried in vacuum to obtain white powdery diblock copolymer PEO-PNIPAAM;
s3, adding the PEO-PNIPAAm-Br, the SPMAP and the PMDETA prepared in the step S2 into a reaction container, adding a mixed solvent E for dissolving, vacuumizing and introducing nitrogen for three times, adding a catalyst under the protection of nitrogen, continuously introducing nitrogen for 10-20 min for sealing, placing the reaction container into an oil bath at 60 ℃, stirring for reaction for 12-20 h, and exposing the reaction container to air after the reaction is finished to terminate the reaction; THF is used as eluent, the reaction mixture is passed through an alkaline alumina column, a large amount of solvent is removed by rotary evaporation, the product is dropped into iced ether for sedimentation and washing for 3 times, and the white powdery triblock copolymer PEO-PNIPAAm-PSPMAP is obtained by filtration and drying in vacuum.
The PEO in the step S1 is PEO2000。
The catalyst added in the step S2 and the step S3 is CuBr.
The molar ratio of the PEO, the BIBB, the ETA and the DMAP added in the step S1 is 1 (2-2.2) to 2-2.2: (2-2.2), the volume ratio of the added mixed liquid B to the added mixed liquid C is 1: (2-3); the molar ratio of the PEO-Br, the NIPAAm, the PMDETA and the catalyst added in the step S2 is 1: (60-80): (1-1.2): (1-1.5); the molar ratio of the PEO-PNIPAAm-Br, the SPMAP, the PMDETA and the catalyst added in the step S3 is 1: (80-100): (1-1.2): (1-1.5).
The mass ratio of the PEO to the organic solvent A in the mixed solution C in the step S1 is 1: (3-5).
The organic solvent A added in the step S1 is dichloromethane DCM; the mixed solvent D added in the step S2 is H2Mixing O and dimethylformamide DMF according to a volume ratio of 1 (1-1.5); the mixed solvent E added in the step S3 is H2O and isopropanol IPA in a volume ratio of 1: (1-1.5) mixing the above components.
The block copolymer is prepared into solution, and the solution is used as coupling agent to form covalent bonding coating which is self-assembled on the inner wall of a quartz capillary to form coating capillary separation protein.
The coated capillary tube is prepared by the following steps:
p1, washing the capillary column for 5min by using NaOH solution with the concentration of 1mol/L, and then standing for 15min to activate the silicon hydroxyl on the inner wall of the capillary column;
p2 sequentially using photosensitive diazo resin aqueous solution, water, block copolymer aqueous solution, water and N2Sequentially washing the capillary columns treated in the step P1 for 5min respectively; the mass concentration ratio of the photosensitive diazo resin to the block copolymer is 1 (1-2);
and P3, repeating the step P2 to recoat the capillary column, placing the capillary column in an oven at the temperature of 60-80 ℃ overnight, or placing the capillary column under an ultraviolet lamp with the wavelength of 365nm for exposure for 20min, and thus constructing the protein adsorption resistant self-cleaning covalent bonding coating.
The capillary column has an effective length of 41cm and an internal diameter of 50 or 75 μm; the diazo resin is diphenylamine-4-diazo resin.
Preferably, the capillary column has an inner diameter of 75 μm.
And (3) heating the coating capillary tube to 38 ℃, and then washing the coating with deionized water for 5min to realize self-cleaning of the coating capillary tube.
Compared with the prior art, the invention has the following beneficial effects:
(1) the protein adsorption resistant self-cleaning block copolymer is synthesized on a macroinitiator through an atom transfer radical polymerization method, each block has good hydrophilicity, the protein adsorption resistant performance is guaranteed, and the temperature-sensitive block part provides a foundation for the self-cleaning performance. The copolymer and the coupling agent are carried out in a water phase in a layer-by-layer self-assembly mode, and the preparation process is simple and quick.
(2) The preparation method of the block copolymer provided by the invention utilizes a simplified atom transfer radical polymerization method: dissolving initiator, monomer and complexing agent in organic solvent, vacuumizing and introducing nitrogen for 3 times, adding catalyst in N2The reaction is carried out under protection, the complex experimental process of three times of liquid nitrogen freezing, vacuumizing, nitrogen introducing and unfreezing circulation is avoided, the reaction under the basic experimental condition is ensured, and the resource is saved.
(3) The invention realizes covalent bonding between sulfonic group of triblock copolymer and silicon hydroxyl on the inner wall of a capillary by photo-curing crosslinking reaction of photosensitive diazo resin, keeps the stability of a coating, and solves the problems of protein adsorption and coating cleaning.
(4) The preparation method of the capillary tube coating provided by the invention comprises the steps of utilizing photosensitive diazo resin to carry out self-assembly and curing crosslinking on a triblock copolymer to form a covalent bonding coating, sequentially flushing a capillary column with diazo resin solution, triblock copolymer solution and distilled water for 5min, assembling and growing the coating on the inner wall of a quartz capillary tube through hydrogen bond action, and after the self-assembly process is finished, pyrolyzing to enable the self-assembly coating and the inner wall of the capillary tube to carry out curing crosslinking to form the covalent bonding coating. The coating can be prepared in a water phase in a self-assembly mode, and the preparation process is simple and quick. The chemical bonding of the coating is realized by utilizing the pyrolysis, curing and crosslinking of diazo resin, so that the energy-intensive or environment-aggressive treatment is avoided, and a simple, rapid and environment-friendly method is provided for realizing the surface modification of the capillary.
(5) The coating on the inner wall of the capillary tube is formed by using diazo resin as a coupling agent and performing pyrolysis, curing and crosslinking, so that a coupling agent silanization reagent with high toxicity and high price is avoided, the process for preparing the capillary tube self-cleaning covalent bonding coating is simplified, the repeatability is good, the raw material price is low, and the production process is green and environment-friendly.
(6) The block copolymer coating has good hydrophilicity and excellent protein adsorption resistance, and the temperature-sensitive block can automatically clean the capillary, so that the cyclic regeneration and use of the coating are facilitated.
Drawings
FIG. 1 is an infrared spectrum of the protein adsorption resistant self-cleaning block copolymer obtained in example 1;
FIG. 2 is a plot of the minimum critical solution temperature (LCST) of the self-cleaning block copolymer against protein adsorption obtained in example 1;
FIG. 3 is a diagram showing the separation of mixed proteins before and after self-cleaning by the capillary tube with the protein adsorption-resistant self-cleaning coating obtained in example 8.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention will be described in further detail below. This invention may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete and will fully convey the concept of the invention to those skilled in the art, and the present invention will only be defined by the appended claims.
The invention provides an anti-protein adsorption self-cleaning block copolymer, which is a triblock copolymer prepared by taking three hydrophilic materials of polyethylene glycol monomethyl ether (PEO), N-isopropylacrylamide (NIPAAm) and 3-sulfopropyl methacrylate (SPMAP) as raw materials and adopting an Atom Transfer Radical Polymerization (ATRP), and the triblock copolymer has the simple formula: PEO-PNIPAAM-PSPMAP, the low critical solution temperature is 33 ℃, and the structural formula is shown as the formula (I).
Figure BDA0002500969850000061
Wherein PEO is an initiation block, NIPAAm is a temperature sensitive block, and SPMAP is a cross-linking block;
in the formula (I), m is an integer of 16-35, and n is an integer of 8-15.
The protein adsorption resistant self-cleaning block copolymer is synthesized on a macroinitiator through an atom transfer radical polymerization method, each block has good hydrophilicity, the protein adsorption resistant performance is guaranteed, and the temperature-sensitive block part provides a foundation for the self-cleaning performance. The copolymer is carried out in a water phase in a layer-by-layer self-assembly mode, and the preparation process is simple and quick. The polymer material is applied to capillary electrophoresis, and is used as a covalent coating through diazo resin crosslinking, so that the stability of the coating is maintained, and the problems of difficult protein adsorption and recycling of the existing capillary wall coating material are solved.
The invention also provides a preparation method of the protein adsorption resistant self-cleaning block copolymer, which comprises the following steps:
s1, under the protection of nitrogen atmosphere, adding 2-bromoisobutyryl bromide (BIBB), triethylamine (ETA) and dimethyl pyridine (DMAP) into a proper amount of organic solvent Dichloromethane (DCM), stirring for dissolving, and then placing in an ice water bath to obtain a mixed solution B; mixing polyethylene glycol monomethyl ether (PEO) and DCM according to the mass ratio 1 (3-5) to obtain a mixed solution C, then dropwise adding the mixed solution C into the mixed solution B, reacting at room temperature for 10-20 h, extracting with NaCl solution for three times, drying an organic phase with anhydrous magnesium sulfate, filtering, performing rotary evaporation to remove the solvent, settling a product in glacial ethyl ether, filtering, and performing vacuum distillationDrying in the air to obtain a faint yellow macromolecular initiator PEO-Br; wherein the PEO is preferably PEO2000；
S2, adding the initiators PEO-Br, NIPAAm and PMDETA prepared in the step S1 into a reaction container, adding a mixed solvent D for dissolving, vacuumizing and introducing nitrogen for three times, adding a catalyst CuBr under the protection of nitrogen, then continuously introducing nitrogen for 10-20 min for sealing, placing the reaction container into an oil bath at 30 ℃, stirring for reacting for 3-5 h, and exposing the reaction container to air after the reaction is finished to terminate the reaction; THF is used as an eluent, the reaction mixture is subjected to alkaline alumina column, a large amount of solvent is removed by rotary evaporation, the product is dropped into ethyl glacial ether for sedimentation and washing for 3 times, and the product is filtered and dried in vacuum to obtain white powdery diblock copolymer PEO-PNIPAAM; wherein the added mixed solvent D is H2And (3) mixing the O and the dimethyl formamide DMF according to the volume ratio of 1 (1-1.5).
S3, adding the PEO-PNIPAAm-Br, the SPMAP and the PMDETA prepared in the step S2 into a reaction container, adding a mixed solvent E for dissolving, vacuumizing and introducing nitrogen for three times, adding a catalyst CuBr under the protection of nitrogen, continuously introducing nitrogen for 10-20 min for sealing, placing the reaction container into an oil bath at 60 ℃, stirring for reaction for 12-20 h, and after the reaction is finished, exposing the reaction container to air to terminate the reaction; THF is used as eluent, the reaction mixture is processed by an alkaline alumina column, a large amount of solvent is removed by rotary evaporation, the product is dripped into ice ether for sedimentation and washing for 3 times, and the white powdery triblock copolymer PEO-PNIPAAm-PSPMAP is obtained by filtration and vacuum drying; wherein the added mixed solvent E is H2O and isopropanol IPA in a volume ratio of 1: (1-1.5) mixing the above components.
The preparation method of the block copolymer provided by the invention utilizes a simplified atom transfer radical polymerization method: dissolving initiator, monomer and complexing agent in organic solvent, vacuumizing and introducing nitrogen for 3 times, adding catalyst in N2The reaction is carried out under protection, the fussy experimental process of liquid nitrogen freezing, vacuumizing, nitrogen-introducing unfreezing circulation for three times is avoided, the reaction under the basic experimental condition is ensured, and the resource is saved.
The invention also provides an application of the anti-protein adsorption self-cleaning block copolymer in coating, the block copolymer is prepared into an aqueous solution, photosensitive diazo resin is used as a coupling agent to form a covalent bonding coating which is self-assembled on the inner wall of a quartz capillary tube to form a coating capillary tube, and the method specifically comprises the following steps:
p1, washing the capillary column with NaOH solution with concentration of 1mol/L for 5min, and standing for 15min to activate the silicon hydroxyl on the inner wall of the capillary column, wherein the capillary column has effective length of 41cm and inner diameter of 50 or 75 μm, preferably 75 μm;
p2, sequentially using photosensitive diazo resin solution, water, block copolymer solution, water and N2Sequentially washing the capillary columns treated in the step P1 for 5min respectively; the mass ratio of the added photosensitive diazo resin to the block copolymer is 1 (1-2), and the diazo resin is preferably diphenylamine-4-diazo resin;
and P3, repeating the step P1 to reactivate the capillary column, placing the capillary column in an oven at the temperature of 60-80 ℃ overnight, or placing the capillary column under an ultraviolet lamp with the wavelength of 365nm for exposure for 20min, and thus constructing the protein adsorption resistant self-cleaning covalent bonding coating.
When the separation effect is poor, the coating capillary is heated to 38 ℃, and then the coating capillary is washed by deionized water for 5min, so that the self-cleaning of the coating capillary can be realized.
The invention realizes covalent bonding between sulfonic group of triblock copolymer and silicon hydroxyl on the inner wall of a capillary by photo-curing crosslinking reaction of photosensitive diazo resin, keeps the stability of a coating, and solves the problems of protein adsorption and coating cleaning.
The preparation method of the capillary tube coating provided by the invention comprises the steps of utilizing photosensitive diazo resin to carry out self-assembly and curing crosslinking on triblock copolymer to form a covalent bonding coating, sequentially flushing a capillary column with diazo resin solution, triblock copolymer solution and distilled water for 5min, carrying out assembly growth on the inner wall of a quartz capillary tube through hydrogen bond action on the coating, and after the self-assembly process is finished, pyrolyzing the self-assembly coating to carry out curing crosslinking on the self-assembly coating and the inner wall of the capillary tube to form the covalent bonding coating. The coating can be prepared in a water phase in a self-assembly mode, and the preparation process is simple and quick. The chemical bonding of the coating is realized by utilizing the pyrolysis, curing and crosslinking of diazo resin, so that the energy-intensive or environment-aggressive treatment is avoided, and a simple, rapid and environment-friendly method is provided for realizing the surface modification of the capillary.
The coating on the inner wall of the capillary tube is formed by using diazo resin as a coupling agent and performing pyrolysis, curing and crosslinking, so that a coupling agent silanization reagent with high toxicity and high price is avoided, the process for preparing the capillary tube self-cleaning covalent bonding coating is simplified, the repeatability is good, the raw material price is low, and the production process is green and environment-friendly.
The block copolymer coating has good hydrophilicity and excellent protein adsorption resistance, and the temperature-sensitive block can automatically clean the capillary, so that the cyclic regeneration and use of the coating are facilitated.
The coating capillary tube is characterized in that under the condition that the temperature is lower than the critical dissolving temperature (preferably 25 ℃), the block copolymer material layer is in a stretching state, the coating is a hydrophilic coating, the protein adsorption resistance of the coating is stronger, and the coating capillary tube is adopted for carrying out electrophoretic separation on protein; when the anti-protein adsorption self-cleaning covalent bonding coating modified on the inner wall of the capillary is polluted, the separation effect is influenced. The polymer material layer is in a contracted state under the condition that the temperature is increased to be higher than the low critical solution temperature of the coating (preferably 38 ℃), so that the contact surface of protein and the coating is reduced, and the self-cleaning of the protein adsorption resistant coating capillary can be realized by flushing the triblock copolymer coating capillary with water.
Example 1
The preparation method of the protein adsorption resistant self-cleaning block copolymer provided by the embodiment comprises the following steps:
s1, adding 1.22g DMAP, 1.24ml BIBB, 1.4ml TEA and 15ml DCM into a 150ml three-neck flask, stirring and dissolving into an ice water bath, and under the protection of nitrogen, dropwise adding 10g PEO dissolved by a constant pressure dropping funnel200050ml of DCM solution was stirred slowly at room temperature for 12 h. After the reaction is finished, extracting the mixture for three times by using NaCl solution, drying the organic phase by using anhydrous magnesium sulfate, filtering, removing the solvent by rotary evaporation, settling the product in glacial ethyl ether, filtering, and drying in vacuum for 12 hours to obtain a light yellow ATRP macroinitiator PEO2000-Br。
S2, 0.5g of PEO as the initiator prepared in the step S12000-Br, 2.2632g NIPAAm, 0.043g PMDETA into a 150ml three-necked flask, 10ml DMF and H2Dissolving the O in a mixed solvent D mixed according to the volume ratio of 3:2, and performing vacuum pumping-nitrogen gas introduction circulation for three times. Adding 0.35g of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 30 ℃, and stirring for reacting for 4 hours. After the reaction was completed, the reaction system was terminated by exposure to air, THF was used as an eluent, the reaction mixture was passed through an alkaline alumina column, much of the solvent was removed by rotary evaporation, the solution was dropped into glacial ethyl ether to be settled and washed 3 times, filtered and dried in a vacuum oven for 12 hours to obtain a white powdery solid diblock copolymer PEO2000-PNIPAAm。
S3, 0.19g of PEO prepared in step S22000-PNIPAAm-Br, 1.231g SPMAP, 0.008g PMDETA into a 150ml three-necked flask, 10ml IPA and H2And stirring and dissolving the O in a mixed solvent E mixed according to the volume ratio of 3:2, and performing vacuum pumping-nitrogen gas introduction circulation for three times. Adding 0.0072g of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 60 ℃, and stirring for reacting for 15 hours. After completion of the reaction, the reaction system was terminated by exposure to air, THF was used as eluent, and the reaction mixture was passed through a basic alumina column. Removing a large amount of solvent by rotary evaporation, dropping the solution into glacial ethyl ether, settling and washing for 3 times, filtering, and drying in a vacuum drying oven for 12h to obtain white powder solid triblock copolymer PEO2000-PNIPAAm-PSPMAP。
FIG. 1 is an infrared spectrum of a triblock copolymer obtained in example 1, 1171cm in the spectrum-1、1539cm-1、1637cm-1Respectively corresponding to a sulfonic group absorption peak on 3-sulfopropyl methacrylate, an amide II band and an amide I band of N-isopropyl acrylamide, and indicating that the synthesis of the invention successfully obtains a triblock copolymer PEO2000-PNIPAAm-PSPMAP. FIG. 2 is a polymer PEO2000-minimum critical solution temperature (LCST) diagram of PNIPAAm-PSPMAP, with a solution permeability decreasing with increasing temperature, with a minimum critical solution temperature of 33 ℃.
Example 2
The preparation method of the protein adsorption resistant self-cleaning block copolymer provided by the embodiment comprises the following steps:
s1, adding 1.22g DMAP, 1.24ml BIBB, 1.4ml TEA and 15ml DCM into a 150ml three-neck flask, stirring and dissolving into an ice-water bath, and under the protection of nitrogen, dropwise adding a dropping funnel with constant pressure and 25g PEO dissolved into the mixture500050ml of DCM solution was stirred slowly at room temperature for 12 h. After the reaction is finished, extracting the mixture for three times by using NaCl solution, drying the organic phase by using anhydrous magnesium sulfate, filtering, removing the solvent by rotary evaporation, settling the product in glacial ethyl ether, filtering, and drying in vacuum for 12 hours to obtain a light yellow ATRP macroinitiator PEO5000-Br。
S2, 1.25g of PEO as the initiator prepared in the step S15000-Br, 2.663g NIPAAm, 0.043g PMDETA into a 150ml three-necked flask, 10ml DMF and H2Dissolving the O in a mixed solvent D mixed according to the volume ratio of 3:2, and performing vacuum pumping-nitrogen gas introduction circulation for three times. Adding 0.42g of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 30 ℃, and stirring for reacting for 4 hours. After the reaction was completed, the reaction system was terminated by exposure to air, THF was used as an eluent, the reaction mixture was passed through an alkaline alumina column, much of the solvent was removed by rotary evaporation, the solution was dropped into ethyl glacial ether to be settled and washed 3 times, filtered and vacuum-dried for 12 hours to obtain a white powdery solid diblock copolymer PEO5000-PNIPAAm。
Polymer PEO5000The minimum critical solution temperature of PNIPAAm is above 50 ℃, and the self-cleaning process is not beneficial to maintaining the activity of protein, so that the PNIPAAm is not selected as an anti-protein adsorption material and does not carry out subsequent reaction.
Example 3
The preparation method of the protein adsorption resistant self-cleaning block copolymer provided by the embodiment comprises the following steps:
s1, adding 0.672g DMAP, 0.744ml BIBB, 0.84ml TEA and 10ml DCM into a 150ml three-neck flask, stirring and dissolving into an ice-water bath, and dropwise adding 10g PEO dissolved in a constant pressure dropping funnel under the protection of nitrogen200030ml of DCM solution (b) are slowly stirred at room temperature for 18 h. Reaction ofAfter the completion, extracting with NaCl solution for three times, drying the organic phase with anhydrous magnesium sulfate, filtering, removing the solvent by rotary evaporation, settling the product in glacial ethyl ether, filtering and drying in vacuum for 12h to obtain light yellow ATRP macroinitiator PEO2000-Br。
S2, 1g of PEO as the initiator prepared in the step S12000-Br, 4.526g NIPAAm, 0.0952g PMDETA were put in a 150ml three-necked flask, and 10ml DMF and H were added2Dissolving the O in the mixed solvent D mixed according to the volume ratio of 1:1, vacuumizing, and introducing nitrogen for three times. 0.0776g of CuBr is added under the protection of nitrogen, nitrogen is introduced for 10min for sealing, the reaction is placed in an oil bath at 30 ℃, and the reaction is stirred for 4 hours. After the reaction was completed, the reaction system was terminated by exposure to air, THF was used as an eluent, the reaction mixture was passed through an alkaline alumina column, much of the solvent was removed by rotary evaporation, the solution was dropped into ethyl glacial ether to be settled and washed 3 times, filtered and vacuum-dried for 12 hours to obtain a white powdery solid diblock copolymer PEO2000-PNIPAAm。
S3, 0.19g of PEO prepared in step S22000-PNIPAAm-Br, 1.231g SPMAP, 0.008g PMDETA into a 150ml three-necked flask, 10ml IPA and H2And stirring and dissolving the mixed solvent E in which the O is mixed according to the volume ratio of 1:1, and performing vacuum pumping-nitrogen gas introduction circulation for three times. Adding 0.0072g of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 60 ℃, and stirring for reacting for 15 hours. After completion of the reaction, the reaction system was terminated by exposure to air, THF was used as eluent, and the reaction mixture was passed through a basic alumina column. Removing a large amount of solvent by rotary evaporation, dropping the solution into glacial ethyl ether, settling and washing for 3 times, filtering and vacuum drying for 12h to obtain white powder solid triblock copolymer PEO2000-PNIPAAm-PSPMAP。
Example 4
The preparation method of the protein adsorption resistant self-cleaning block copolymer provided by the embodiment comprises the following steps:
s1, adding 1.22g DMAP, 1.24ml BIBB, 1.4ml TEA and 20ml DCM into a 150ml three-neck flask, stirring and dissolving into an ice-water bath, and under the protection of nitrogen, dropwise adding 10g PEO dissolved in a constant pressure dropping funnel2000DCM solution of (1)50ml, stirred slowly at room temperature for 12 h. After the reaction is finished, extracting three times by NaCl solution, drying the organic phase by anhydrous magnesium sulfate, filtering, removing the solvent by rotary evaporation, settling the product in ice ether, filtering and drying in vacuum for 12 hours to obtain a light yellow ATRP macroinitiator PEO2000-Br。
S2, 0.5g of PEO as the initiator prepared in the step S12000adding-Br, 1.697g NIPAAm and 0.0516g PMDETA into a 150ml three-neck flask, adding 10ml DMF and H2Dissolving the O in a mixed solvent D mixed according to the volume ratio of 3:2, and performing vacuum pumping-nitrogen gas introduction circulation for three times. Adding 0.525g of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 30 ℃, and stirring for reaction for 4 hours. After the reaction was completed, the reaction system was terminated by exposure to air, THF was used as an eluent, the reaction mixture was passed through an alkaline alumina column, a large amount of the solvent was removed by rotary evaporation, the solution was dropped into iced ether to be settled and washed 3 times, filtered and vacuum-dried for 12 hours, to obtain a white powdery solid diblock copolymer PEO 2000-PNIPAAm.
S3, 0.19g of PEO prepared in step S22000-PNIPAAm-Br, 1.231g SPMAP, 0.0104g PMDETA into a 150ml three-necked flask, adding 10ml of a mixed solvent E prepared by mixing isopropanol IPA and H2O in a volume ratio of 3:2, stirring and dissolving, and vacuumizing and circulating with nitrogen for three times. 0.01076g of CuBr is added under the protection of nitrogen, nitrogen is introduced for 10min for sealing, the reaction is placed in an oil bath at 60 ℃, and the reaction is stirred for 15 hours. After completion of the reaction, the reaction system was terminated by exposure to air, THF was used as eluent, and the reaction mixture was passed through a basic alumina column. Removing a large amount of solvent by rotary evaporation, dropping the solution into glacial ethyl ether, settling and washing for 3 times, filtering and vacuum drying for 12h to obtain white powder solid triblock copolymer PEO2000-PNIPAAm-PSPMAP。
Example 5
The preparation method of the protein adsorption resistant self-cleaning block copolymer provided by the embodiment comprises the following steps:
s1, adding 1.22g DMAP, 1.24ml BIBB, 1.4ml TEA and 20ml DCM into a 150ml three-neck flask, stirring and dissolving into an ice-water bath, and under the protection of nitrogen, dropwise adding 10g PEO dissolved in a constant pressure dropping funnel200050ml of DCM solution was stirred slowly at room temperature for 12 h. After the reaction is finished, extracting the mixture for three times by using NaCl solution, drying the organic phase by using anhydrous magnesium sulfate, filtering, removing the solvent by rotary evaporation, settling the product in glacial ethyl ether, filtering and drying the product in vacuum for 12 hours to obtain a light yellow ATRP macroinitiator PEO2000-Br。
S2, 0.5g of PEO as the initiator prepared in the step S12000-Br, 2.8588g NIPAAm, 0.043g PMDETA into a 150ml three-necked flask, 10ml DMF and H2Dissolving the O in a mixed solvent D mixed according to the volume ratio of 3:2, and performing vacuum pumping-nitrogen gas introduction circulation for three times. Adding 0.35g of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 30 ℃, and stirring for reacting for 4 hours. After the reaction is finished, the reaction system is stopped by exposing to air, the reaction mixture is directly added into a dialysis bag with the molecular weight of 3500, deionized water is used for dialysis for 3-5 days until the dialysate is colorless and clear, and the white flocculent solid diblock copolymer PEO which is freeze-dried is used2000-PNIPAAm。
S3, 0.19g PEO prepared by step S22000-PNIPAAm-Br, 1.231g SPMAP, 0.008g PMDETA into a 150ml three-necked flask, 10ml IPA and H2And stirring and dissolving the O in a mixed solvent E mixed according to the volume ratio of 3:2, and performing vacuum pumping-nitrogen gas introduction circulation for three times. Adding 0.0072g of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 60 ℃, and stirring for reacting for 15 hours. After the reaction is finished, the reaction system is stopped by exposing to air, the reaction mixture is directly added into a dialysis bag with the molecular weight of 5000, deionized water is used for dialysis for 3-5 days until the dialysate is colorless and clear, and the white flocculent solid triblock copolymer PEO is freeze-dried2000-PNIPAAm-PSPMAP。
Example 6
The preparation method of the protein adsorption resistant self-cleaning block copolymer provided by the embodiment comprises the following steps:
s1, adding 0.01mol DMAP, 0.01mol BIBB, 0.01mol TEA and 15ml DCM into a three-neck flask, stirring and dissolving into an ice water bath, and dropwise adding 0.005mol PEO dissolved in a constant pressure dropping funnel under the protection of nitrogen200030ml of DCM solution in roomStirring slowly at room temperature for 18 h. After the reaction is finished, extracting the mixture for three times by using NaCl solution, drying the organic phase by using anhydrous magnesium sulfate, filtering, removing the solvent by rotary evaporation, settling the product in glacial ethyl ether, filtering and drying the product in vacuum for 12 hours to obtain a light yellow ATRP macroinitiator PEO2000-Br。
S2, adding 0.0005mol of PEO as the initiator prepared in the step S12000adding-Br, 0.03mol NIPAAm and 0.0005mol PMDETA into a three-neck flask, adding 10ml DMF and H2Dissolving the O in the mixed solvent D in the volume ratio of 1:1.5, and circulating for three times by vacuumizing and introducing nitrogen. Adding 0.0005mol of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 30 ℃, and stirring for reacting for 4 hours. After the reaction was completed, the reaction system was terminated by exposure to air, THF was used as an eluent, the reaction mixture was passed through an alkaline alumina column, much of the solvent was removed by rotary evaporation, the solution was dropped into ethyl glacial ether to be settled and washed 3 times, filtered and vacuum-dried for 12 hours to obtain a white powdery solid diblock copolymer PEO2000-PNIPAAm。
S3, 0.0001mol of PEO prepared in the step S22000-PNIPAAm-Br, 0.008mol SPMAP, 0.0001mol PMDETA into a three-neck flask, 10ml IPA and H2And stirring and dissolving the O in a mixed solvent E mixed according to the volume ratio of 1:1.5, and performing vacuum pumping-nitrogen gas introduction circulation for three times. Adding 0.0001mol of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 60 ℃, and stirring for reaction for 15 hours. After completion of the reaction, the reaction system was terminated by exposure to air, THF was used as eluent, and the reaction mixture was passed through a basic alumina column. Removing a large amount of solvent by rotary evaporation, dropping the solution into glacial ethyl ether, settling and washing for 3 times, filtering and vacuum drying for 12h to obtain white powder solid triblock copolymer PEO2000-PNIPAAm-PSPMAP。
Example 7
The preparation method of the protein adsorption resistant self-cleaning block copolymer provided by the embodiment comprises the following steps:
s1, adding 0.011mol DMAP, 0.011mol BIBB, 0.011mol TEA and 15ml DCM into a three-neck flask, stirring and dissolving into an ice water bath, and using a constant-pressure liquid drop drain to drain under the protection of nitrogen0.005mol of PEO dissolved in the solution is dripped into a bucket200045ml of DCM solution were slowly stirred at room temperature for 15 h. After the reaction is finished, extracting the mixture for three times by using NaCl solution, drying the organic phase by using anhydrous magnesium sulfate, filtering, removing the solvent by rotary evaporation, settling the product in glacial ethyl ether, filtering and drying in vacuum to obtain light yellow ATRP macromolecular initiator PEO2000-Br。
S2, adding 0.0005mol of PEO as the initiator prepared in the step S12000adding-Br, 0.04mol NIPAAm and 0.0006mol PMDETA into a three-neck flask, adding 8ml DMF and H2Dissolving the O in a mixed solvent D mixed according to the volume ratio of 1:1.5, vacuumizing, and introducing nitrogen for three times. Adding 0.00075mol of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 30 ℃, and stirring for reaction for 4 hours. After the reaction was completed, the reaction system was terminated by exposure to air, THF was used as an eluent, the reaction mixture was passed through an alkaline alumina column, much of the solvent was removed by rotary evaporation, the solution was dropped into glacial ethyl ether to be settled and washed 3 times, filtered and vacuum-dried to obtain white powdery solid diblock copolymer PEO2000-PNIPAAm。
S3, 0.0001mol PEO prepared by the step S22000-PNIPAAm-Br, 0.01mol SPMAP, 0.00012mol PMDETA into a three-neck flask, 8ml IPA and H2And stirring and dissolving the O in a mixed solvent E mixed according to the volume ratio of 1:1.5, and performing vacuum pumping-nitrogen gas introduction circulation for three times. Adding 0.00015mol of CuBr under the protection of nitrogen, introducing nitrogen for 10min, sealing, placing the reaction in an oil bath at 60 ℃, and stirring for reaction for 15 hours. After completion of the reaction, the reaction system was terminated by exposure to air, THF was used as eluent, and the reaction mixture was passed through a basic alumina column. Removing a large amount of solvent by rotary evaporation, dropping the solution into glacial ethyl ether for settling and washing for 3 times, filtering and vacuum drying to obtain white powder solid triblock copolymer PEO2000-PNIPAAm-PSPMAP。
Example 8
In the application of the self-cleaning block copolymer with protein adsorption resistance in the coating aspect, the triblock copolymer PEO obtained in example 1 is used2000Preparing a 2mg/ml solution from-PNIPAAm-PSPMAP, and using photosensitive diazo resin as a coupling agentThe capillary with covalent bonding coating specifically comprises the following steps:
p1, washing the capillary column with NaOH solution with the concentration of 1mol/L for 5min, and standing for 15min to activate the silicon hydroxyl on the inner wall of the capillary column, wherein the effective length of the capillary column is 41cm, and the inner diameter is 75 μm;
p2, sequentially using photosensitive diazo resin solution, water, triblock copolymer solution, water and N2Sequentially washing the capillary columns treated in the step P1 for 5min respectively; wherein the mass ratio of the added photosensitive diazo resin to the triblock copolymer is 1: 2;
and P3, repeating the step P2 to reactivate the capillary column, and placing the capillary column in an oven with the temperature of 70 ℃ overnight to construct the protein adsorption resistant self-cleaning covalent bonding coating.
When the separation effect is poor, the coating capillary is heated to 38 ℃, and then the coating capillary is washed by deionized water for 5min, so that the self-cleaning of the coating capillary can be realized.
FIG. 3 is a separation diagram of the covalently bonded coated hybrid protein obtained in example 8 and a later self-cleaning characterization diagram thereof, and it can be seen from FIG. 3 that the self-cleaning operation of the coating is performed at 38 ℃ under the condition of keeping the protein activity temperature, so that the recycling of the coated capillary can be effectively realized.
Example 9
In the application of the self-cleaning block copolymer with protein adsorption resistance in the coating aspect, the triblock copolymer PEO obtained in example 1 is used2000-PNIPAAm-PSPMAP is prepared into a solution of 2mg/ml, photosensitive diazo resin is used as a coupling agent to form a covalently bonded coating capillary, and the method specifically comprises the following steps:
p1, washing the capillary column for 30min by using NaOH solution with the concentration of 1mol/L, and activating the silicon hydroxyl on the inner wall of the capillary column, wherein the effective length of the capillary column is 41cm, and the inner diameter of the capillary column is 50 mu m;
p2, sequentially using photosensitive diazo resin solution, water, triblock copolymer solution, water and N2Sequentially washing the capillary columns treated in the step P1 for 5min respectively; wherein the mass ratio of the added photosensitive diazo resin to the triblock copolymer is 1: 1;
and P3, repeating the step P2 to reactivate the capillary column, and placing the capillary column in an oven with the temperature of 60 ℃ overnight to construct the protein adsorption resistant self-cleaning covalent bonding coating.
The capillary column with the inner diameter of 50 mu m becomes smaller after being coated, which is more likely to cause protein adsorption, thus leading the electrophoretic separation effect to be worse, being not beneficial to the subsequent self-cleaning test, so the capillary column is not selected as a coating column.
Example 10
In the application of the self-cleaning block copolymer with protein adsorption resistance in the coating aspect, the triblock copolymer PEO obtained in example 5 is used2000-PNIPAAm-PSPMAP is prepared into a solution of 2mg/ml, photosensitive diazo resin is used as a coupling agent to form a covalently bonded coating capillary, and the method specifically comprises the following steps:
p1, washing the capillary column for 30min by using NaOH solution with the concentration of 1mol/L, and activating the silicon hydroxyl on the inner wall of the capillary column, wherein the effective length of the capillary column is 41cm, and the inner diameter is 75 mu m;
p2, sequentially using photosensitive diazo resin solution, water, triblock copolymer solution, water and N2Sequentially washing the capillary columns treated in the step P1 for 5min respectively; wherein the mass ratio of the added photosensitive diazo resin to the triblock copolymer is 1: 1;
and P3, repeating the step P1 to reactivate the capillary column, and exposing the capillary column under an ultraviolet lamp with the wavelength of 365nm for 20min to construct the protein adsorption resistant self-cleaning covalent bonding coating.
When the separation effect is poor, the coating capillary is heated to 38 ℃, and then the coating capillary is washed by deionized water for 5min, so that the self-cleaning of the coating capillary can be realized.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are intended to be within the scope of the invention.

Claims (10)

1. The protein adsorption resistant self-cleaning block copolymer is characterized in that the block copolymer is a triblock copolymer PEO-PNIPAAm-PSPMAP prepared by taking three hydrophilic materials of polyethylene glycol monomethyl ether PEO, N-isopropyl acrylamide NIPAAm and 3-sulfopropyl methacrylate SPMAP as raw materials through Atom Transfer Radical Polymerization (ATRP), and the structural formula of the triblock copolymer PEO-PNIPAAm-PSPMAP is shown as the formula (I):
Figure 369961DEST_PATH_IMAGE001
（Ⅰ）
wherein PEO is an initiation block, NIPAAm is a temperature sensitive block, and SPMAP is a cross-linking block;
in the formula (I), m is an integer of 16-35, and n is an integer of 8-15.
2. The protein adsorption resistant self-cleaning block copolymer of claim 1, characterized in that the low critical solution temperature of the block copolymer is 33 ℃.
3. A method for preparing the protein adsorption resistant self-cleaning block copolymer as claimed in claim 1 or 2, characterized by comprising the steps of:
s1, under the protection of nitrogen atmosphere, adding 2-bromoisobutyryl bromide (BIBB), triethylamine (ETA) and dimethyl pyridine (DMAP) into a proper amount of organic solvent A, stirring to dissolve, and placing in an ice water bath to obtain a mixed solution B; dissolving polyethylene glycol monomethyl ether (PEO) in a proper amount of organic solvent A, mixing to obtain a mixed solution C, then dropwise adding the mixed solution C into the mixed solution B, reacting at room temperature for 10-20 h, extracting with NaCl solution for three times, drying an organic phase with anhydrous magnesium sulfate, filtering, removing the solvent by rotary evaporation, settling a product in glacial ethyl ether, filtering, and drying in vacuum to obtain a faint yellow macromolecular initiator (PEO-Br);
s2, adding the initiators PEO-Br, NIPAAm and PMDETA prepared in the step S1 into a reaction vessel, adding a mixed solvent D for dissolving, vacuumizing and introducing nitrogen for three times, adding a catalyst under the protection of nitrogen, continuously introducing nitrogen for 10-20 min for sealing, placing the reaction vessel into an oil bath at 30 ℃, stirring for reacting for 3-5 h, and exposing the reaction vessel to air after the reaction is finished so as to terminate the reaction; THF is used as an eluent, the reaction mixture is subjected to alkaline alumina column, a large amount of solvent is removed by rotary evaporation, the product is dropped into ethyl glacial ether for sedimentation and washing for 3 times, and the product is filtered and dried in vacuum to obtain white powdery diblock copolymer PEO-PNIPAAM-Br;
s3, adding the PEO-PNIPAAm-Br, SPMAP and PMDETA prepared in the step S2 into a reaction container, adding a mixed solvent E for dissolving, vacuumizing and introducing nitrogen for three times, adding a catalyst under the protection of nitrogen, continuously introducing nitrogen for 10-20 min for sealing, placing the reaction container into an oil bath at 60 ℃, stirring for reacting for 12-20 h, and introducing air to terminate the reaction after the reaction is finished; THF is used as eluent, the reaction mixture is passed through an alkaline alumina column, a large amount of solvent is removed by rotary evaporation, the product is dropped into ethyl glacial ether for sedimentation and washing for 3 times, and the white powdery triblock copolymer PEO-PNIPAAm-PSPMAP is obtained after filtration and drying in vacuum.
4. The method for preparing a protein adsorption-resistant self-cleaning block copolymer as claimed in claim 3, wherein the PEO in the step S1 is PEO2000(ii) a The catalyst added in the step S2 and the step S3 is CuBr.
5. The method for preparing the protein adsorption resistant self-cleaning block copolymer as claimed in claim 3, wherein the molar ratio of the PEO, the BIBB, the ETA and the DMAP added in the step S1 is 1 (2-2.2) to (2-2.2): (2-2.2), the volume ratio of the added mixed liquid B to the added mixed liquid C is 1: (2-3); the molar ratio of the PEO-Br, the NIPAAm, the PMDETA and the catalyst added in the step S2 is 1: (60-80): (1-1.2): (1-1.5); the molar ratio of the PEO-PNIPAAm-Br, the SPMAP, the PMDETA and the catalyst added in the step S3 is 1: (80-100): (1-1.2): (1-1.5).
6. Preparation of the protein adsorption resistant self-cleaning block copolymer according to claim 3The method is characterized in that the organic solvent A added in the step S1 is dichloromethane DCM; the mixed solvent D added in the step S2 is H2A mixture of O and dimethylformamide DMF according to a volume ratio of 1 (1-1.5); the mixed solvent E added in the step S3 is H2O and isopropanol IPA in a volume ratio of 1: (1-1.5).
7. The use of the self-cleaning block copolymer with protein adsorption resistance as claimed in claim 1 or 2, wherein the block copolymer is prepared into a solution, and a covalent bonding coating is formed by using photosensitive diazo resin as a coupling agent, and the covalent bonding coating is self-assembled on the inner wall of a quartz capillary to form a coating capillary separation protein.
8. Use of a protein adsorption resistant self-cleaning block copolymer as claimed in claim 7 for coating, characterized in that the coated capillary is prepared by the following steps:
p1, washing the capillary column for 5min by using NaOH solution with the concentration of 1mol/L, and then standing for 15min to activate the silicon hydroxyl on the inner wall of the capillary column;
p2, sequentially using photosensitive diazo resin aqueous solution, water, block copolymer aqueous solution, water and N2Sequentially washing the capillary columns treated in the step P1 for 5min respectively; the mass concentration ratio of the photosensitive diazo resin to the block copolymer is 1 (1-2);
and P3, repeating the step P2 to recoat the capillary column, placing the capillary column in an oven at the temperature of 60-80 ℃ overnight, or placing the capillary column under an ultraviolet lamp with the wavelength of 365nm for exposure for 20min, and thus constructing the protein adsorption resistant self-cleaning covalent bonding coating.
9. Use of the protein adsorption resistant self-cleaning block copolymer in coatings according to claim 8, characterized in that the capillary column has an effective length of 41cm and an internal diameter of 50 or 75 μ ι η; the diazo resin is diphenylamine-4-diazo resin.
10. The use of the protein adsorption resistant self-cleaning block copolymer in coating according to claim 7, wherein the self-cleaning of the coating capillary is achieved by rinsing the coating with deionized water for 5min after the coating capillary is heated to 38 ℃.
CN202010432143.5A 2020-05-20 2020-05-20 Anti-protein-adsorption self-cleaning block copolymer, preparation method and application thereof Active CN111533865B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010432143.5A CN111533865B (en) 2020-05-20 2020-05-20 Anti-protein-adsorption self-cleaning block copolymer, preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010432143.5A CN111533865B (en) 2020-05-20 2020-05-20 Anti-protein-adsorption self-cleaning block copolymer, preparation method and application thereof

Publications (2)

Publication Number Publication Date
CN111533865A CN111533865A (en) 2020-08-14
CN111533865B true CN111533865B (en) 2022-07-05

Family

ID=71972753

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010432143.5A Active CN111533865B (en) 2020-05-20 2020-05-20 Anti-protein-adsorption self-cleaning block copolymer, preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN111533865B (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116217792A (en) * 2022-12-03 2023-06-06 西北工业大学 Preparation method of super-hydrophilic material and durable wetting gradient surface

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007078819A2 (en) * 2005-12-16 2007-07-12 Arkema Inc. Low surface energy block co-polymer preparation methods and applications
CN104262555A (en) * 2014-10-22 2015-01-07 山东交通学院 Block polymer with multi-response property for temperature and carbon dioxide and preparation method thereof

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101022879A (en) * 2001-03-13 2007-08-22 戴斯－分析公司 Heat and moisture exchange device

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007078819A2 (en) * 2005-12-16 2007-07-12 Arkema Inc. Low surface energy block co-polymer preparation methods and applications
CN104262555A (en) * 2014-10-22 2015-01-07 山东交通学院 Block polymer with multi-response property for temperature and carbon dioxide and preparation method thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Bio-Inspired Design and Fabrication of Micro/Nano-Brush Dual Structural Surfaces for Switchable Oil Adhesion and Antifouling;Tao Du;《Small》;20171231;1602020 *
共价键合毛细管电泳涂层柱的高效制备与应用;陈垚;《中国优秀博硕士学位论文全文数据库（硕士）工程科技I辑》;20190115;B014-781 *
聚(N-异丙基丙烯酰胺)基共聚物结构对抗蛋白吸附性能的影响;刘娜;《中国组织工程研究与临床康复》;20090917;第13卷(第38期);7442-7446 *

Also Published As

Publication number Publication date
CN111533865A (en) 2020-08-14

Similar Documents

Publication Publication Date Title
CN108484825B (en) Reversible light-operated hydrophobic azobenzene fluorine-containing copolymer and membrane preparation method thereof
CN106905158B (en) Benzophenone type molecule for directly modifying substrate and application thereof
CN102580574B (en) Antipollution self-cleaning polyvinylidene fluoride membrane and method for producing same
CN112375191B (en) Block copolymer, preparation method and application thereof
CN107629379B (en) Reversible light-operated hydrophobic composite membrane of one kind and preparation method thereof
CN105884988B (en) The adjustable high perforation hexagonal columnar film of P4VP-b-PMA (Az) block copolymer, column diameter and preparation method
CN102952236A (en) Molecularly imprinted polymeric microsphere resin applicable to aqueous solution system and preparing method thereof
CN106497302A (en) Photocatalytic self-cleaning parents' network marine anti-pollution coating and preparation method thereof
CN110652890B (en) Surface hydrophilic modification method for polyacrylonitrile separation membrane
CN111533865B (en) Anti-protein-adsorption self-cleaning block copolymer, preparation method and application thereof
CN102952245A (en) Environment-friendly preparation method of multi-hydroxyl polymer molecular brush
CN104628974B (en) A kind of amphipathic copolymer and preparation method thereof assigning membrane material pH responses
CN105001413B (en) A kind of preparation method of polyethylene glycol diazonium polymer
CN106496464B (en) A kind of amphipathic network of UV light anti-fouling type and preparation method thereof
Li et al. Biofiber waste derived zwitterionic and photocatalytic dye adsorbent: Switchable selectivity, in-situ degradation and multi-tasking application
CN102617770B (en) Preparation method of linkage polymer based on cucurbituril [6]
CN105566587A (en) Preparation method and application of polyvinyl alcohol diazo polymer
CN104558543A (en) Modification method of silica gel
CN107434851B (en) Preparation method of photosensitive hydrogel
CN104592532A (en) Amphiphilic copolymer network and preparation method thereof
CN104892873B (en) Polymer of multi-functional amphipathic low-surface-energy and preparation method and application
CN110372836B (en) Vinyl sulfone surface initiated aqueous phase anion polymerization method
CN112876617B (en) Preparation method of porous molecularly imprinted sustained-release material
JP6027972B2 (en) Compounds, polymers, film coatings, and medical devices
CN110156615B (en) Compound DA-I2959, Polymer DA-Polymer, preparation method and use method thereof, and oil lubricating material

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant