CN113372608B - PNIPAAm (AM)/DA composite imprinted gel and preparation method and application thereof - Google Patents

PNIPAAm (AM)/DA composite imprinted gel and preparation method and application thereof Download PDF

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CN113372608B
CN113372608B CN202110538529.9A CN202110538529A CN113372608B CN 113372608 B CN113372608 B CN 113372608B CN 202110538529 A CN202110538529 A CN 202110538529A CN 113372608 B CN113372608 B CN 113372608B
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pnipaam
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gel
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lysozyme
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CN113372608A (en
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柯渔
范家琛
敖宁建
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Jinan University
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Abstract

The invention belongs to the field of biological functional materials, and discloses PNIPAAm (AM)/DA composite imprinted gel, and a preparation method and application thereof. The preparation method introduces Polydopamine (PDA) into temperature-sensitive poly N-isopropylacrylamide (PNIPAAm) and Acrylamide (AM) composite gel in a free radical polymerization mode to prepare novel PNIPAAm (AM)/DA composite imprinted gel, and because PDA is introduced into the original PNIPAAm (AM) imprinted layer, the strength of the novel composite imprinted gel is greatly improved while the temperature-sensitive characteristic of PNIPAAm is kept, so that the damage of surface three-dimensional imprinted pores of the composite imprinted gel is avoided in a time period of temperature-responsive change, and the temperature-sensitive controlled release performance of the composite imprinted gel on template protein is finally improved.

Description

PNIPAAm (AM)/DA composite imprinted gel and preparation method and application thereof
Technical Field
The invention belongs to the field of biological functional materials, and particularly relates to PNIPAAm (AM)/DA composite imprinted gel, and a preparation method and application thereof.
Background
Biological macromolecules play a very important role in our living organisms, and continuous exploration of macromolecules such as peptides, proteins and the like not only can provide basic scientific basis for explaining the activity rule of substances in the organisms, but also provides necessary basic theoretical basis and solution for overcoming difficult and complicated diseases in the medical field in the future. When the biomedical material used by people enters into the body and plays a role, the first reaction is the nonspecific adhesion of macromolecular substances existing in blood or body fluid and the surface of the biological material, then different influences are generated on the interaction of the material and cell tissues in the body due to different types and quantities of adsorbed proteins, and finally a series of immune reactions are shown on a macroscopic level, so that the research is clear on how the macromolecular substances and the surface of the material interact, and the biomedical material has very important basic research value and invisible practical value in future clinical application. (Wu Z, Chen H, Liu X, et al. protein amplification on poly (N-vinylpyrrolidone) -modified silicon substrates prepared by surface-induced atom transfer, polymerization.2009; 25: 2900-6).
Since the nineties of the last century, the development and application of molecular imprinting technology has focused essentially on the area of design and preparation around the development of small molecules. After many years of continuous exploration and innovation, the molecular imprinting technology has been widely applied to many fields, including the fields of biomedical materials, biosensors, drug delivery and controlled release, etc. However, the development of the molecular imprinting polymer, which is particularly applicable to the biomedical field, is slow, so that the preparation of the low-cost molecular imprinting material with excellent structure and performance has important research value, the molecular imprinting material is firstly simple to prepare, has good specific identification performance on target macromolecules, can keep certain stability under some harsh conditions, and can meet the requirements to the maximum extent by using the molecular imprinting technology. (Miao X M CZY, Ma H Y, Li Z B, Xue N Wang P.A Novel and Label-free Platform for MicroRNA Detection Based on the Fluorescence queuing of positional Charge Gold Nanoparticles to Ag nanocrusters, Analytical chemistry 2017; 90: 1098-.
In the field of biomaterials, high molecular polymer materials having intelligent responsiveness have been receiving attention of researchers, and poly (N-isopropylacrylamide) (PNIPAAm) as a representative of high molecular materials can respond to external temperature changes, and further has a lower critical transition temperature LCST close to the body temperature of a human body. At present, poly (N-isopropylacrylamide) (PNIPAAm) has been applied to surface modification of various substrates in combination with molecular imprinting technology to prepare intelligent biomaterials with controlled release of macromolecular substances such as proteins (Itoga K, Okano T. the high functional release of temperature-responsive culture media for metabolic advanced cell housing engineering. journal of Materials Chemistry).
2010；20:8768-75.)。
The poly (N-isopropylacrylamide) (PNIPAAm) hydrogel can generate phase transition behavior under the stimulation of temperature, so that the poly (N-isopropylacrylamide) (PNIPAAm) hydrogel is expected to be applied to load fixation, controlled release and cyclic absorption of target active substances.
Although PNIPAAm has the characteristic of temperature response, the LCST of the PNIPAAm is close to the body temperature (Cho EC, Kim DH, Cho K. contacts of oils on soluble substrates in aqueous media: correction with AFM data on protein addition, Langmuir.2008; 24:9974-8), after the PNIPAAm is prepared into a temperature-sensitive type system and is swelled, permanent collapse and damage are easy to generate due to the notch sensitivity of the PNIPAAm, and the performance of controlling release of the PNIPAAm is finally reduced. Therefore, in recent years, research in the scientific field has been mainly focused on the improvement of the thermal response and mechanical properties of poly (N-isopropylacrylamide) (PNIPAAm).
Disclosure of Invention
In order to overcome the technical defects of the poly (N-isopropylacrylamide) (PNIPAAm) as an intelligent response material and according to the temperature-sensitive characteristics and application characteristics of the poly (N-isopropylacrylamide) (PNIPAAm), the invention mainly aims to provide a preparation method of PNIPAAm (AM)/DA composite imprinted gel. According to the method, Polydopamine (PDA) is introduced into poly (N-isopropyl acrylamide) (PNIPAAm) gel and Acrylamide (AM) so as to form intermolecular hydrogen bonds in a gel network, and the mechanical properties and the protein load controlled release performance of the finally prepared gel and an imprinting layer are improved. The whole reaction process is simple, the operation is convenient, and the prepared gel and the imprinted coating are uniform and stable and brown in color.
The second purpose of the invention is to provide PNIPAAm (AM)/DA composite blotting gel prepared by the method.
The third purpose of the invention is to provide the application of the PNIPAAm (AM)/DA composite blotting gel.
The primary purpose of the invention is realized by the following technical scheme:
a preparation method of PNIPAAm (AM)/DA composite imprinting gel comprises the following steps:
(1) dissolving Dopamine (DA) in an aqueous solution with the pH value of 8.0-8.5, and stirring and oxidizing in air to form a polydopamine solution;
(2) sequentially adding N-isopropylacrylamide, Acrylamide (AM), N-methylenebisacrylamide and lysozyme into the polydopamine solution prepared in the step (1), introducing nitrogen, exhausting oxygen in the mixed solution, adding an initiator ammonium persulfate, introducing nitrogen again, and stirring in a closed manner to form a pre-polymerization solution;
(3) adding N, N, N ', N' -tetramethylethylenediamine into the prepolymerization liquid in the step (2), and isolating oxygen for reaction to prepare a PNIPAAm (AM)/DA composite imprinted gel initial product;
(4) and (4) eluting the PNIPAAm (AM)/DA composite blotting gel primary product obtained in the step (3) in NaCI solution, and eluting lysozyme template protein on the surface of the primary product to obtain a PNIPAAm (AM)/DA composite blotting gel final product.
Preferably, the dopamine, N-isopropylacrylamide, acrylamide, N, N-methylenebisacryloyl, lysozyme, ammonium persulfate, N, N, N ', N' -tetramethylethylenediamine and the aqueous solution with the pH value of 8.0-8.5 are mixed according to the mass-to-volume ratio of 1.5-2.5 mg: 301.18-310 mg: 20-28 mg: 15-20 mg: 10 mg: 20-25 mg: 18-25 μ L: 5ml of the solution was prepared.
Preferably, the dopamine, N-isopropylacrylamide, acrylamide, N, N-methylenebisacryloyl, lysozyme, ammonium persulfate, N, N, N ', N' -tetramethylethylenediamine and the aqueous solution with the pH value of 8.0-8.5 are mixed according to the mass-to-volume ratio of 1.5-2.5 mg: 301.18 mg: 26.19 mg: 18 mg: 10 mg: 20 mg: 20 μ L of: 5ml of the solution was prepared.
Preferably, the dopamine, N-isopropylacrylamide, acrylamide, N, N-methylenebisacrylamide, lysozyme, ammonium persulfate, N, N, N ', N' -tetramethylethylenediamine and the aqueous solution with the pH value of 8.0-8.5 are mixed according to a mass-to-volume ratio of 1.5 mg: 301.18 mg: 26.19 mg: 18 mg: 10 mg: 20 mg: 20 μ L of: 5ml of the solution was prepared.
Preferably, the dopamine, N-isopropylacrylamide, acrylamide, N, N-methylenebisacrylamide, lysozyme, ammonium persulfate, N, N, N ', N' -tetramethylethylenediamine and the aqueous solution with the pH value of 8.0-8.5 are mixed according to a mass-to-volume ratio of 2.0 mg: 301.18 mg: 26.19 mg: 18 mg: 10 mg: 20 mg: 20 μ L of: 5ml of the solution was prepared.
Preferably, the dopamine, N-isopropylacrylamide, acrylamide, N, N-methylenebisacrylamide, lysozyme, ammonium persulfate, N, N, N ', N' -tetramethylethylenediamine and the aqueous solution with the pH value of 8.0-8.5 are mixed according to a mass-to-volume ratio of 2.5 mg: 301.18 mg: 26.19 mg: 18 mg: 10 mg: 20 mg: 20 μ L of: 5ml of the solution was prepared.
Preferably, the oxidation time in step (1) is 0.5 h.
Preferably, the time for the first nitrogen gas introduction in the step (2) is 0.5h, and the time for the second nitrogen gas introduction is 15 min.
Preferably, the stirring temperature of the dopamine solution in the step (2) is 25-45 ℃, and the stirring speed is 100-150 r/min.
Preferably, the specific processing steps of step (4) are: and (3) transferring the PNIPAAm (AM)/DA composite imprinting gel prepared in the step (3) between two cleaned glass slides by using a pipette gun, removing the upper glass slide after the reaction is finished, shaking the lysozyme molecular imprinting layer in NaCI solution with the mass concentration of 0.9% by a shaking table, eluting lysozyme template protein on the surface, and preparing a PNIPAAm (AM)/DA composite imprinting gel final product.
Preferably, the reaction time is 6 hours, and the shaking time of the shaking table is 12 hours.
The second purpose of the invention is realized by the following technical scheme:
PNIPAAm (AM)/DA composite blotting gel prepared by the method.
The third purpose of the invention is realized by the following technical scheme:
application of PNIPAAm (AM)/DA composite imprinted gel in biomedical materials
Preferably, the applications include applications in the fields of enzyme immobilization, drug controlled release, separation, and cyclic absorption.
The mechanism of the invention is as follows: when the temperature of the traditional PNIPAAm (AM) gel is changed (during heating), the amide group originally combined with water molecules and forming hydrogen bonds in the PNIPAAm is broken to form intramolecular hydrogen bonds, the polymer is shrunk due to the pulling of the acting force of the hydrogen bonds formed in the molecule, and the gel is possibly broken in the shrinking process.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the PNIPAAm (AM)/DA composite imprinted gel obtained by the method has the temperature response characteristic, can control and release protein macromolecules loaded on the surface of an imprinted layer by adjusting the temperature, has better mechanical property and cannot be easily damaged like the traditional PNIPAAm hydrogel in the phase change process;
(2) the PNIPAAm (AM)/DA composite imprinted gel obtained by the method is 0.3-0.5 mm;
(3) the PNIPAAm (AM)/DA composite imprinted gel obtained by the method is uniform and stable.
Drawings
FIG. 1 is a diagram of the appearance of PNIPAAm (AM)/DA composite imprinted gel prepared in example 1, example 2 and example 3 before and after temperature rise at 25 ℃ and 45 ℃;
FIG. 2 is a graph of the appearance and morphology of the PNIAAm (AM) conventional non-imprinted complex gel prepared in comparative example 1 at 25 deg.C (left) and 45 deg.C (right);
FIG. 3 is a bar graph of the adsorption performance of PNIPAAm (AM)/DA complex blotting gels prepared in example 1, example 2 and example 3 on lysozyme protein;
FIG. 4 is a bar graph showing the adsorption of PNIPAAm (AM)/DA composite blotting gel prepared in example 2 at a PDA content of 0.4g/L to lysozyme at different temperatures;
FIG. 5 is an IR spectrum of PDA, PNIPAAm (AM)/DA in example 2;
fig. 6 is a graph showing the compression performance test of the gel prepared in example 2.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
The reagents used in the examples are commercially available without specific reference.
Example 1: preparation of PNIPAAm (AM)/DA composite blotting gel
(1) 1.5mg dopamine (from Sigma) was dissolved in 5ml aqueous solution pH 8.5 and subsequently the dopamine solution was oxidised in air with stirring for 0.5h to form a polydopamine solution.
(2) 301.18mg of N-isopropylacrylamide, 26.19mg of acrylamide, 18mg of N, N-methylenebisacrylamide and 10mg of lysozyme were dissolved in the polydopamine solution, and then the mixture was stirred under sealed conditions and purged with nitrogen for 0.5 h. After nitrogen is introduced to exhaust oxygen in the solution, 20mg of initiator ammonium persulfate is added, then nitrogen is introduced again for 15min, and after the nitrogen introduction is finished, a pre-polymerization solution is formed; finally, 20 mu L N of N, N ', N' -tetramethylethylenediamine is added into the solution to prepare a PNIPAAm (AM)/DA composite imprinted gel initial product; and (3) quickly transferring the gel liquid between two cleaned glass slides by using a liquid transfer gun, removing the upper glass slide after 6 hours of reaction, shaking the lysozyme molecular imprinting layer in NaCI solution with the mass concentration of 0.9% for 12 hours in a shaking table, and eluting lysozyme template protein on the surface to prepare a PNIPAAm (AM)/DA composite imprinting gel final product.
Example 2: preparation of PNIPAAm (AM)/DA composite blotting gel
(1) 2.0mg dopamine (from Sigma) was dissolved in 5ml aqueous solution pH 8.5 and subsequently the dopamine solution was oxidised in air with stirring for 0.5h to form a polydopamine solution.
(2) 301.18mg of N-isopropylacrylamide, 26.19mg of acrylamide, 18mg of N, N-methylenebisacrylamide and 10mg of lysozyme were dissolved in the polydopamine solution, and then the mixture was stirred under sealed conditions and purged with nitrogen for 0.5 h. After nitrogen is introduced to exhaust oxygen in the solution, 20mg of initiator ammonium persulfate is added, then nitrogen is introduced again for 15min, and after the nitrogen introduction is finished, a pre-polymerization solution is formed; finally, 20 mu L N of N, N ', N' -tetramethylethylenediamine is added into the solution to prepare a PNIPAAm (AM)/DA composite imprinted gel initial product; and (3) quickly transferring the gel liquid between two cleaned glass slides by using a liquid transfer gun, removing the upper glass slide after 6 hours of reaction, shaking the lysozyme molecular imprinting layer in NaCI solution with the mass concentration of 0.9% for 12 hours in a shaking table, and eluting lysozyme template protein on the surface to prepare a PNIPAAm (AM)/DA composite imprinting gel final product. FIG. 4 is a bar graph showing the adsorption of prepared PNIPAAm (AM)/DA composite imprinted gel with PDA content of 0.4g/L to lysozyme at different temperatures in the present example, wherein the larger the IF (imprinting factor) value, the higher the Western blotting efficiency of the composite imprinted layer, the MIP represents imprinted gel, and the NIP represents non-imprinted gel, so that it can be seen from FIG. 4 that the gel has obvious temperature response characteristics to the lysozyme protein adsorption capacity; as shown in fig. 5, which is an infrared spectrum diagram of PDA, PNIPAAm (AM)/DA in this embodiment, it can be seen that the characteristic absorption peaks of the PNIPAAm (AM)/DA composite gel substantially cover the characteristic absorption peaks of PDA and PNIPAAm (AM), indicating that the PNIPAAm (AM)/DA composite gel has been successfully synthesized. FIG. 6 shows the compression property test chart of the composite gel prepared in this example. As can be seen from FIG. 6, with the introduction of PDA, the mechanical properties of PNIPAAm (AM)/DA composite gel are also improved to a certain extent.
Example 3: preparation of PNIPAAm (AM)/DA composite blotting gel
(1) 2.5mg dopamine (from Sigma) was dissolved in 5ml aqueous solution pH 8.5 and subsequently the dopamine solution was oxidised in air with stirring for 0.5h to form a polydopamine solution.
(2) 301.18mg of N-isopropylacrylamide, 26.19mg of acrylamide, 18mg of N, N-methylenebisacrylamide and 10mg of lysozyme were dissolved in the polydopamine solution, and then the mixture was stirred under sealed conditions and purged with nitrogen for 0.5 h. After nitrogen is introduced to exhaust oxygen in the solution, a pre-polymerization solution is formed; adding 20mg of initiator ammonium persulfate, then introducing nitrogen for 15min again, and finally adding 20 mu L N of N, N ', N' -tetramethylethylenediamine into the solution after the nitrogen introduction is finished to prepare a PNIPAAm (AM)/DA composite imprinted gel initial product; and (3) quickly transferring the gel liquid between two cleaned glass slides by using a liquid transfer gun, removing the upper glass slide after 6 hours of reaction, shaking the lysozyme molecular imprinting layer in NaCI solution with the mass concentration of 0.9% for 12 hours in a shaking table, and eluting lysozyme template protein on the surface to prepare a PNIPAAm (AM)/DA composite imprinting gel final product.
Comparative example 1: preparation of PNIPAAm (AM) traditional non-blotting composite gel
301.18mg of N-isopropylacrylamide, 26.19mg of acrylamide, 18mg of N, N-methylenebisacrylamide and 10mg of lysozyme were dissolved in deionized water, and then the mixture was stirred under sealed conditions and purged with nitrogen for 0.5 h. After nitrogen is introduced to exhaust oxygen in the solution, 20mg of initiator ammonium persulfate is added, then nitrogen is introduced again for 15min, after nitrogen introduction is finished, 20 mu L N of N, N ', N' -tetramethyl ethylenediamine is finally added into the solution, the gel liquid is rapidly transferred between two cleaned slides by using a liquid transfer gun, after 6 hours of reaction, the upper slide is removed, the gel is shaken in a NaCI solution with the mass concentration of 0.9% for 12 hours, and the preparation of PNIPAAm (AM) traditional non-imprinted composite gel is completed. Figure 2 shows the appearance and appearance of the PNIAAm (AM) traditional non-imprinted complex gel prepared in the comparative example at 25 deg.C (left) and 45 deg.C (right).
Comparative example 2: preparation of PNIPAAm (AM)/DA non-blotting composite gel
(1) 2.0mg dopamine (from Sigma) was dissolved in 5ml aqueous solution pH 8.5 and subsequently the dopamine solution was oxidised in air with stirring for 0.5h to form a polydopamine solution.
(2) 301.18mg of N-isopropylacrylamide, 26.19mg of acrylamide and 18mg of N, N-methylenebisacrylamide were dissolved in the polydopamine solution, and then the mixture was stirred with a closed state and purged with nitrogen for 0.5 h. After nitrogen is introduced to exhaust oxygen in the solution, 20mg of initiator ammonium persulfate is added, then nitrogen is introduced again for 15min, after nitrogen introduction is finished, 20 mu L N of N, N ', N' -tetramethyl ethylenediamine is finally added into the solution, the gel liquid is rapidly transferred between two cleaned glass slides by using a liquid transfer gun, after reaction is carried out for 6h, the upper glass slide is removed, the gel is shaken in a NaCI solution with the mass concentration of 0.9% for 12h, and the preparation of PNIPAAm (AM)/DA traditional non-imprinted composite gel is completed.
FIG. 1 shows the morphology of the PNIPAAm (AM)/DA complex imprinting gel prepared in example 1, example 2 and example 3 before and after temperature rise at 25 ℃ and 45 ℃; figure 2 shows the appearance and appearance of the PNIAAm (AM) traditional non-imprinted complex gel prepared in comparative example 1 at 25 ℃ (left) and 45 ℃ (right); from fig. 1, it can be seen that the novel composite blotting gel is in macroscopic contrast with the pnipaam (am) conventional non-blotting gel of fig. 2, which causes volume change due to temperature change;
FIG. 3 is a bar graph showing the adsorption performance of PNIPAAm (AM)/DA complex blotting gels prepared in examples 1, 2 and 3 on lysozyme protein; as can be seen from the figure, all three composite blotting gels maintain certain adsorption capacity on lysozyme protein, and the blotting efficiency of example 2 is highest.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims (9)

1. A preparation method of PNIPAAm (AM)/DA composite imprinting gel is characterized by comprising the following steps:
(1) dissolving dopamine in an aqueous solution with the pH value of 8.0-8.5, and stirring and oxidizing in air to form a polydopamine solution;
(2) sequentially adding N-isopropylacrylamide, acrylamide, N-methylenebisacrylamide and lysozyme into the polydopamine solution prepared in the step (1), introducing nitrogen, exhausting oxygen in the mixed solution, adding an initiator ammonium persulfate, introducing nitrogen again, and stirring in a closed manner to form a pre-polymerization solution;
(3) adding N, N, N ', N' -tetramethylethylenediamine into the prepolymerization liquid in the step (2), and isolating oxygen for reaction to prepare a PNIPAAm (AM)/DA composite imprinted gel initial product;
(4) eluting the PNIPAAm (AM)/DA composite imprinted gel primary product obtained in the step (3) in a NaCl solution, and eluting lysozyme template protein on the surface of the primary product to prepare a PNIPAAm (AM)/DA composite imprinted gel final product;
the dopamine, N-isopropyl acrylamide, N, N-methylene diacryloyl, lysozyme, ammonium persulfate, N, N, N ', N' -tetramethyl ethylenediamine and the aqueous solution with the pH value of 8.0-8.5 are mixed according to the mass-to-volume ratio of 1.5-2.5 mg: 301.18-310 mg: 20-28 mg: 15-20 mg: 10 mg: 20-25 mg: 18-25 μ L: 5ml of the solution was prepared.
2. The PNIPAAm (AM)/DA composite imprinted gel according to claim 1, wherein the dopamine, N-isopropylacrylamide, acrylamide, N, N-methylenebisacrylamide, lysozyme, ammonium persulfate, N, N, N ', N' -tetramethylethylenediamine, and the aqueous solution having a pH of 8.0 to 8.5 are mixed in a mass-to-volume ratio of 1.5 to 2.5 mg: 301.18 mg: 26.19 mg: 18 mg: 10 mg: 20 mg: 20 μ L of: 5ml of the solution was prepared.
3. The PNIPAAm (AM)/DA composite imprinted gel according to claim 1, wherein the dopamine, N-isopropylacrylamide, acrylamide, N, N-methylenebisacrylamide, lysozyme, ammonium persulfate, N, N, N ', N' -tetramethylethylenediamine, and the aqueous solution having a pH of 8.0 to 8.5 are mixed in a mass-to-volume ratio of 1.5 mg: 301.18 mg: 26.19 mg: 18 mg: 10 mg: 20 mg: 20 μ L of: 5ml of the solution was prepared.
4. The method for preparing PNIPAAm (AM)/DA composite blotting gel according to claim 1, wherein the time for the first nitrogen gas injection in step (2) is 0.5h, and the time for the second nitrogen gas injection is 15 min.
5. The preparation method of PNIPAAm (AM)/DA composite imprinted gel according to claim 1, wherein the stirring temperature of the dopamine solution in the step (2) is 25-45 ℃, and the stirring speed is 100-150 r/min.
6. The preparation method of PNIPAAm (AM)/DA composite blotting gel according to claim 1, wherein the specific processing steps of the step (4) are as follows: and (3) transferring the PNIPAAm (AM)/DA composite imprinting gel prepared in the step (3) between two cleaned slides by using a pipette gun, removing the upper slide after the reaction is finished, shaking the lysozyme molecular imprinting layer in a NaCl solution with the mass concentration of 0.9% by using a shaking table, and eluting lysozyme template protein on the surface to prepare a PNIPAAm (AM)/DA composite imprinting gel final product.
7. PNIPAAm (AM)/DA composite blotting gel prepared by the preparation method according to any one of claims 1 to 6.
8. Use of the PNIPAAm (AM)/DA composite blotting gel according to claim 7 in biomedical materials.
9. The PNIPAAm (AM)/DA composite imprinted gel according to claim 8, wherein the application comprises the application in the fields of enzyme immobilization, drug controlled release, separation and cyclic absorption.
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