CN106824307B - mixed anion exchange medium and preparation method thereof - Google Patents
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Abstract
The invention discloses a mixed anion exchange medium and a preparation method thereof, and the method comprises the following steps: s1, carrying out ring-opening etherification reaction on epoxy chloropropane and phenol to form ligand phenyl glycidyl ether, and catalyzing by using a phase transfer catalyst PEG to obtain the phenyl glycidyl ether in one step; s2, reacting epichlorohydrin with trimethylamine hydrochloride to obtain ligand 2, 3-epoxypropyl trimethyl ammonium chloride; s3, replacing agarose with acetone aqueous solutions with different proportions, and crosslinking the synthesized ligand phenyl glycidyl ether to the surface of the agarose; and then crosslinking the synthesized phenyl agarose beads with 2, 3-epoxypropyltrimethylammonium chloride to obtain the anion exchange medium. The medium has the characteristics of hydrophobic interaction and strong anion exchange mode, improves the selectivity to target protein, can be applied to various fields in the biological pharmacy, particularly to the preparation of monoclonal antibodies, and has obvious effect on removing host DNA, host protein, polymers and viruses.
Description
Technical Field
the invention belongs to the technical field of biochemical engineering and high polymer materials, and particularly relates to a mixed anion exchange medium and a preparation method thereof.
Background
The biological separation medium is a biological separation material which is recognized at present and has the most promising development prospect, is widely used for producing enzymes with important medical values, growth factors, vaccines, monoclonal antibodies and other biological pharmaceutical industries, and becomes a key technology of the medical biological high-technology industry.
ion Exchange Chromatography (IEC) is a highly efficient separation method that relies mainly on charge-charge interactions to perform separation using small differences in charge in charged molecules, and has high separation capacity. Because the ion exchange chromatography has high resolution, large working capacity and easy operation, the ion exchange chromatography is widely applied to the fields of medicine, chemical industry, metallurgy, food and the like, and also becomes an important method for separating and purifying protein, polypeptide, nucleic acid and nucleic acid. The ion exchange medium is composed of three parts: (1) a crosslinked network backbone, i.e., a parent structure; (2) functional groups fixed on the framework, which are charged groups, mark the basic performance of the ion exchange chromatography medium; (3) mobile, exchangeable mobile ions (which may be referred to as counterions) of opposite charge to the functional groups. The anion exchange media commonly used in the market at present mainly include conventional anion exchange media: a positively charged compound Diethylaminoethyl (DEAE) as a ligand DEAE chromatographic medium and a positively charged compound quaternary ammonium salt (Q) as a ligand Q chromatographic medium, such as DEAE Sepharose FF and QSepharose FF of GE; mixed mode anion exchange media such as Capto adhere; mixed mode chromatography media refers to ligands that bind two or more different separation matrices on a filler matrix, i.e. bind different ion exchange, affinity, size exclusion and hydrophobic modes. Therefore, the modes of separating the proteins can be combined and utilized, the selectivity of purification is improved, impurities are removed, and the process is simplified.
Disclosure of Invention
In view of the above-mentioned deficiencies in the prior art, the present invention provides a mixed anion exchange medium and a method for preparing the same. The mixed anion exchange medium (Bestarose MIX-A DL) has the characteristics of hydrophobic interaction and strong anion exchange mode, can improve the selectivity of target protein, and simplifies the process.
In order to achieve the purpose, the invention adopts the following technical scheme:
A preparation method of a mixed type anion exchange medium comprises the following steps:
S1, synthesis of ligand phenyl glycidyl ether: performing ring-opening etherification reaction on epoxy chloropropane and phenol to form phenyl glycidyl ether, and catalyzing by using a phase transfer catalyst polyethylene glycol to obtain the phenyl glycidyl ether in one step;
S2, synthesis of ligand 2, 3-epoxypropyltrimethylammonium chloride: under the alkaline condition, reacting epichlorohydrin with trimethylamine hydrochloride to obtain 2, 3-epoxypropyltrimethylammonium chloride (GTA);
S3, crosslinking of ligand and agarose: firstly, gradually replacing agarose beads with acetone aqueous solution with volume fractions of 20 vol%, 40 vol%, 60 vol%, 80 vol% and 100 vol%, and crosslinking the synthesized ligand phenyl glycidyl ether to the surface of agarose; and then crosslinking the synthesized phenyl agarose beads with 2, 3-epoxypropyl trimethyl ammonium chloride to obtain the mixed anion exchange medium Bestarose MIX-A DL.
Preferably, in the synthesis of phenyl glycidyl ether in step S1: mixing phenol, epoxy chloropropane, PEG and trichloromethane, heating to 35-45 ℃, and reacting for 10 hours; after the reaction is finished, filtering, washing with water, distilling under reduced pressure of 4mmHg, and collecting a product of fraction at the temperature of 93-95 ℃; then detecting the content of the product by gas chromatography for later use. In the technical scheme, the phase transfer catalyst PEG is adopted, so that the etherification reaction is favorably carried out.
further, the molar ratio of the phenol to the epichlorohydrin is 1: 2; the volume ratio of the PEG to the chloroform solvent is 1: 10.
preferably, in the synthesis of ligand 2, 3-epoxypropyltrimethylammonium chloride in step S2: putting a certain amount of epoxy chloropropane into a reactor, cooling to 0-5 ℃, continuously stirring, adding a certain mass of trimethylamine hydrochloride, and reacting for 3-5 hours at normal temperature; and after the reaction is finished, filtering, washing for 3 to 5 times by using acetone, and drying in vacuum to obtain the GTA white solid.
Further, the molar ratio of the trimethylamine hydrochloride to the epichlorohydrin is 1: (4-6).
Further, the molar ratio of the trimethylamine hydrochloride to the epichlorohydrin is 1: 5.
Preferably, the crosslinking of the ligand and the agarose in step S3 comprises:
s31, phenyl-containing agarose bead synthesis: crosslinking phenyl glycidyl ether with agarose beads, taking 100ml of crosslinked beads with the agarose content of 6 wt%, loading into a chromatographic column, loading into the chromatographic column, replacing water in the beads with acetone aqueous solutions with different proportions, gradually adjusting the volume fractions of the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally passing through the column with 100 vol% acetone; pouring the replaced agarose beads into a reactor, adding 5ml of acetone and 0.3ml of BF3 diethyl ether solution, and uniformly stirring; then adding 5ml of acetone solution with 0.4M of phenyl glycidyl ether, and stirring for 1.5h at the temperature of 20 ℃ to finish the reaction; firstly, washing with acetone, then replacing acetone in the beads with acetone aqueous solutions with different proportions, gradually adjusting the volume fractions of the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally washing with water to obtain phenyl-containing agarose beads;
S32, MIX-A mixed mode medium synthesis: the agarose beads containing phenyl are crosslinked with 2, 3-epoxypropyltrimethylammonium chloride (GTA), 100ml of agarose beads containing phenyl are poured into a reactor, 100ml of 50 wt% NaOH is added, the mixture is fully mixed, GTA is added, the mixture is continuously stirred, the reaction is stopped after the mixture reacts for 3 hours at the temperature of 50 ℃, ethanol and deionized water are alternately used for washing for 3 times, and finally the product is obtained and stored in 20 wt% of ethanol.
the invention also provides a mixed anion exchange medium Bestarose MIX-A DL prepared by the preparation method.
The structure of the mixed anion exchange medium Bestarose MIX-A DL is shown as a formula (I),
The above-mentionedagarose beads are indicated.
The invention has the beneficial effects that:
1) the mixed anion exchange medium is prepared by reacting epoxy chloropropane phenol to obtain phenyl glycidyl ether, reacting epoxy chloropropane with trimethylamine hydrochloride to obtain 2, 3-epoxypropyl trimethyl ammonium chloride, and crosslinking the phenyl glycidyl ether and the 2, 3-epoxypropyl trimethyl ammonium chloride to the surface of agarose step by step under alkaline conditions (as shown in figure 1). The medium has strong anion exchange group (quaternary ammonium salt) and hydrophobic group (phenyl) crosslinked on the agarose microsphere, so that the medium has the characteristics of hydrophobic interaction and strong anion exchange mode, improves the selectivity of target protein, and further optimizes the process. According to the characteristics of the mixed type anion exchange medium, the mixed type anion exchange medium can be applied to various fields in the biological pharmacy, particularly in the preparation of monoclonal antibodies, and has remarkable effect on removing host DNA, host protein, polymers and viruses.
2) According to the preparation method, the phase transfer catalyst polyethylene glycol (PEG) is used for catalysis in the synthesis of the ligand phenyl glycidyl ether, so that the glycidyl ether can be obtained in one step, and the yield is improved.
3) The aglucone crosslinked by the epoxy group of the phenyl glycidyl ether and the agarose beads has the advantages of good stability and low aglucone shedding rate, which is very important for the properties of the medium and the product quality, and effectively ensures the application prospect of the medium.
Drawings
FIG. 1 is a flow chart of the preparation method of the present invention.
Detailed Description
the technical solution of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. The following examples are only for illustrating the technical solutions of the present invention more clearly, and the protection scope of the present invention is not limited thereby.
the preparation process of the invention is shown in the flow chart of figure 1.
example 1
S1, Synthesis of ligand 2, 3-epoxypropyltrimethylammonium chloride (GTA): putting 78.3ml of epoxy chloropropane into a reactor, cooling to 0 ℃, continuously stirring, adding 19.12g of trimethylamine hydrochloride (the molecular weight is 95.6) (the molar ratio of the trimethylamine hydrochloride to the epoxy chloropropane is 1: 5), and then reacting for 4 hours at normal temperature; after the reaction is finished, filtering, washing for 3 to 5 times by using acetone, and drying in vacuum to obtain a GTA white solid product, wherein the yield is 97 percent, and the purity is 96 percent;
S2, phenyl glycidyl ether synthesis: mixing 88ml of phenol, 157ml of epoxy chloropropane, 20ml of PEG and 2000ml of trichloromethane, heating to 40 ℃, and reacting for 10 hours; after the reaction is finished, filtering, washing with water, distilling under reduced pressure of 4mmHg, and collecting a product of fraction at the temperature of 93-95 ℃; detecting the content of the product by using gas chromatography for later use;
S3, phenyl-containing agarose bead synthesis: crosslinking phenyl glycidyl ether with agarose beads, loading 100ml of crosslinked beads with agarose content of 6 wt% into a chromatographic column, replacing water in the beads with acetone aqueous solutions with different proportions, gradually adjusting the volume fractions of the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally passing through the column with acetone; pouring the replaced agarose beads into a reactor, adding 5ml of acetone and 0.3ml of BF3 diethyl ether solution, and uniformly stirring; then adding 5ml of acetone solution with 0.4M of phenyl glycidyl ether, and stirring for 1.5h at the temperature of 20 ℃ to finish the reaction; firstly, washing with acetone, then replacing acetone in the beads with acetone aqueous solutions with different proportions, gradually adjusting the volume fractions of the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally washing with water to obtain phenyl-containing agarose beads;
s4, MIX-A mixed mode medium synthesis: the agarose beads containing phenyl and 2, 3-epoxypropyl trimethyl ammonium chloride (GTA) are crosslinked, 100ml of the agarose beads containing phenyl are poured into a reactor, 100ml of 50 wt% NaOH is added, the mixture is fully mixed, GTA is added, the mixture is continuously stirred, the reaction is stopped after 3 hours of reaction at the temperature of 50 ℃, ethanol and deionized water are alternately used for washing for 3 times, finally, the product mixed anion exchange medium Bestarose MIX-A DL is obtained, and the product is placed in 20 wt% ethanol for storage.
Example 2
s1, Synthesis of ligand 2, 3-epoxypropyltrimethylammonium chloride (GTA): putting 78.3ml of epoxy chloropropane into a reactor, cooling to 0 ℃, continuously stirring, adding 23.9g of trimethylamine hydrochloride (the molar ratio of the trimethylamine hydrochloride to the epoxy chloropropane is 1: 4), and then reacting for 4 hours at normal temperature; after the reaction is finished, filtering, washing for 3 to 5 times by using acetone, and drying in vacuum to obtain a GTA white solid product, wherein the yield is 97 percent, and the purity is 96 percent;
S2, phenyl glycidyl ether synthesis: mixing 88ml of phenol, 157ml of epoxy chloropropane, 20ml of PEG and 2000ml of trichloromethane, heating to 40 ℃, and reacting for 10 hours; after the reaction is finished, filtering, washing with water, distilling under reduced pressure of 4mmHg, and collecting a product of fraction at the temperature of 93-95 ℃; detecting the content of the product by using gas chromatography for later use;
s3, phenyl-containing agarose bead synthesis: crosslinking phenyl glycidyl ether with agarose beads, loading 100ml of crosslinked beads with agarose content of 6 wt% into a chromatographic column, replacing water in the beads with acetone aqueous solutions with different proportions, gradually adjusting the volume fractions of the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally passing through the column with acetone; pouring the replaced agarose beads into a reactor, adding 5ml of acetone and 0.3ml of BF3 diethyl ether solution, and uniformly stirring; then adding 5ml of acetone solution with 0.4M of phenyl glycidyl ether, and stirring for 1.5h at the temperature of 20 ℃ to finish the reaction; firstly, washing with acetone, then replacing acetone in the beads with acetone aqueous solutions with different proportions, gradually adjusting the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally washing with water to obtain phenyl-containing agarose beads;
s4, MIX-A mixed mode medium synthesis: the agarose beads containing phenyl and 2, 3-epoxypropyl trimethyl ammonium chloride (GTA) are crosslinked, 100ml of the agarose beads containing phenyl are poured into a reactor, 100ml of 50 wt% NaOH is added, the mixture is fully mixed, GTA is added, the mixture is continuously stirred, the reaction is stopped after 3 hours of reaction at the temperature of 50 ℃, ethanol and deionized water are alternately used for washing for 3 times, finally, the product mixed anion exchange medium Bestarose MIX-A DL is obtained, and the product is placed in 20 wt% ethanol for storage.
example 3
s1, Synthesis of ligand 2, 3-epoxypropyltrimethylammonium chloride (GTA): putting 78.3ml of epoxy chloropropane into a reactor, cooling to 0 ℃, continuously stirring, adding 15.93g of trimethylamine hydrochloride (the molar ratio of the trimethylamine hydrochloride to the epoxy chloropropane is 1: 6), and then reacting for 4 hours at normal temperature; after the reaction is finished, filtering, washing for 3 to 5 times by using acetone, and drying in vacuum to obtain a GTA white solid product, wherein the yield is 97 percent, and the purity is 96 percent;
S2, phenyl glycidyl ether synthesis: mixing 88ml of phenol, 157ml of epoxy chloropropane, 20ml of PEG and 2000ml of trichloromethane, heating to 40 ℃, and reacting for 10 hours; after the reaction is finished, filtering, washing with water, distilling under reduced pressure of 4mmHg, and collecting a product of fraction at the temperature of 93-95 ℃; detecting the content of the product by using gas chromatography for later use;
S3, phenyl-containing agarose bead synthesis: crosslinking phenyl glycidyl ether with agarose beads, taking 100ml of crosslinked beads with the agarose content of 6 wt%, loading into a chromatographic column, loading into the chromatographic column, replacing water in the beads with acetone aqueous solutions with different proportions, gradually adjusting the volume fractions of the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally passing through the column with 100 vol% acetone; pouring the replaced agarose beads into a reactor, adding 5ml of acetone and 0.3ml of BF3 diethyl ether solution, and uniformly stirring; then adding 5ml of acetone solution with 0.4M of phenyl glycidyl ether, and stirring for 1.5h at the temperature of 20 ℃ to finish the reaction; firstly, washing with acetone, then replacing acetone in the beads with acetone aqueous solutions with different proportions, gradually adjusting the volume fractions of the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally washing with water to obtain phenyl-containing agarose beads;
S4, MIX-A mixed mode medium synthesis: the agarose beads containing phenyl and 2, 3-epoxypropyl trimethyl ammonium chloride (GTA) are crosslinked, 100ml of the agarose beads containing phenyl are poured into a reactor, 100ml of 50 wt% NaOH is added, the mixture is fully mixed, GTA is added, the mixture is continuously stirred, the reaction is stopped after 3 hours of reaction at the temperature of 50 ℃, ethanol and deionized water are alternately used for washing for 3 times, finally, the product mixed anion exchange medium Bestarose MIX-A DL is obtained, and the product is placed in 20 wt% ethanol for storage.
example 4
S1, Synthesis of ligand 2, 3-epoxypropyltrimethylammonium chloride (GTA): 783ml of epoxy chloropropane is put into a reactor, is cooled to 0 ℃, is continuously stirred, is added with 191.2g of trimethylamine hydrochloride (the molar ratio of the trimethylamine hydrochloride to the epoxy chloropropane is 1: 5), and then reacts for 4 hours at normal temperature; after the reaction is finished, filtering, washing for 3 to 5 times by using acetone, and drying in vacuum to obtain a GTA white solid product, wherein the yield is 97 percent, and the purity is 96 percent;
S2, phenyl glycidyl ether synthesis: 880ml of phenol, 1570mol of epoxy chloropropane, 200ml of PEG and 20L of trichloromethane are mixed, heated to 35 ℃ and reacted for 10 hours; after the reaction is finished, filtering, washing with water, distilling under reduced pressure of 4mmHg, and collecting a product of fraction at the temperature of 93-95 ℃; detecting the content of the product by using gas chromatography for later use;
S3, phenyl-containing agarose bead synthesis: crosslinking phenyl glycidyl ether with agarose beads, replacing water in the beads with acetone aqueous solutions with different proportions, gradually adjusting the volume fractions of the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally passing through a column with 100 vol% acetone; pouring the replaced agarose beads into a reactor, adding 5ml of acetone and 0.3ml of BF3 diethyl ether solution, and uniformly stirring; then adding 5ml of acetone solution with 0.4M of phenyl glycidyl ether, and stirring for 1.5h at the temperature of 20 ℃ to finish the reaction; firstly, washing with acetone, then replacing acetone in the beads with acetone aqueous solutions with different proportions, gradually adjusting the volume fractions of the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally washing with water to obtain phenyl-containing agarose beads;
s4, MIX-A mixed mode medium synthesis: the agarose beads containing phenyl and 2, 3-epoxypropyl trimethyl ammonium chloride (GTA) are crosslinked, 1000ml of the agarose beads containing phenyl are poured into a reactor, 1000ml of 50 wt% NaOH is added, the mixture is fully mixed, GTA is added, the mixture is continuously stirred, the reaction is stopped after 3 hours of reaction at the temperature of 50 ℃, ethanol and deionized water are alternately used for washing for 3 times, finally, the product mixed anion exchange medium Bestarose MIX-A DL is obtained, and the product is placed in 20 wt% ethanol for storage.
The density of epichlorohydrin in the above examples of the invention was 1.1812 g/mL.
The structure of the mixed anion exchange medium Bestarose MIX-A DL prepared in the embodiment is shown as the formula (I),
The above-mentionedAgarose beads are indicated.
adsorption Effect of Bestarose MIX-A DL on different proteins in the above examples
the adsorption effect of Bestarose MIX-A DL on different proteins was characterized by using the 10% dynamic loading of Bestarose MIX-A DL on Bovine Serum Albumin (BSA), recombinant human serum albumin (r-HSA) and monoclonal antibody (human IgG) against the Bestarose MIX-A DL product prepared in the above example.
TABLE 1 dynamic 10% Loading of Bestarose MIX-A DL and Q Bestarose FF for different proteins in example 1
The adsorption capacity of the medium to the protein is characterized by a dynamic loading of 10% under the condition of no sodium chloride (NaCl). Under the same binding conditions (without NaCl), the 10% dynamic loading of Q Bestarose FF and Bestarose MIX-A DL are almost the same, and the 10% dynamic loading of BSA, r-HSA and human IgG is more than 50 mg/ml.
Under the condition of salt concentration in the presence of 0.1M NaCl, the dynamic loading of BSA (10%) is that Q Bestarose FF (23 mg/ml) is reduced by over 59%, while Bestarose MIX-A DL (47 mg/ml) is over 2 times of the loading of Q Bestarose FF (less than 10% compared with the loading without NaCl); in the presence of a salt concentration of 0.1M NaCl; for 10% of dynamic loading capacity of r-HSA, Q Bestarose FF is 19mg/ml and is reduced by more than 60%, while Bestarose MIX-A DL is 46mg/ml and is more than 2 times of the loading capacity of Q Bestarose FF, and is reduced by less than 10% compared with the condition without NaCl; in the presence of a salt concentration of 0.1M NaCl, Bestarose FF hardly bound to human IgG, whereas Bestarose MIX-A DL hardly changed the dynamic load of 10% to human IgG. The results show that: the combination condition of Bestarose MIX-A DL on protein adsorption can tolerate salt concentration with certain concentration, especially human IgG, so that Bestarose MIX-A DL can be widely applied to antibody drugs.
TABLE 2 dynamic 10% Loading of Bestarose MIX-A DL and Q Bestarose FF for different proteins in example 2
TABLE 3 dynamic 10% Loading of Bestarose MIX-A DL and Q Bestarose FF for different proteins in example 3
TABLE 4 10% dynamic Loading of Bestarose MIX-A DL and Q Bestarose FF for different proteins in example 4
The results, combined with tables 2 to 4, also show that: the binding condition of Bestarose MIX-A DL on protein adsorption can tolerate the salt concentration of a certain concentration, particularly for human IgG, so that the Bestarose MIX-A DL can be widely applied to antibody drugs.
The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.
Claims (8)
1. A preparation method of a mixed type anion exchange medium is characterized by comprising the following steps:
S1, synthesis of ligand phenyl glycidyl ether: performing ring-opening etherification reaction on epoxy chloropropane and phenol to form phenyl glycidyl ether, and catalyzing by using a phase transfer catalyst polyethylene glycol to obtain the phenyl glycidyl ether in one step;
S2, synthesis of ligand 2, 3-epoxypropyltrimethylammonium chloride: reacting epichlorohydrin with trimethylamine hydrochloride to obtain 2, 3-epoxypropyltrimethylammonium chloride GTA;
s3, crosslinking of a ligand and an agarose bead: firstly, gradually replacing agarose beads with 20 vol%, 40 vol%, 60 vol%, 80 vol% and 100 vol% acetone aqueous solution, and crosslinking synthesized ligand phenyl glycidyl ether to the surface of the agarose beads; then crosslinking the synthesized phenyl agarose beads with 2, 3-epoxypropyl trimethyl ammonium chloride to obtain the mixed anion exchange medium;
In the step S3, the crosslinking of the ligand and the agarose beads comprises:
S31, phenyl-containing agarose bead synthesis: crosslinking phenyl glycidyl ether with agarose beads, taking 100ml of crosslinked beads with the agarose content of 6 wt%, loading into a chromatographic column, replacing water in the beads with acetone aqueous solutions with different proportions, gradually adjusting the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally passing through the column with 100 vol% acetone; the displaced agarose beads were poured into a reactor, 5ml of acetone and 0.3ml of BF were added3 stirring the ether solution uniformly; then adding 5ml of acetone solution with 0.4M of phenyl glycidyl ether, and stirring for 1.5h at the temperature of 20 ℃ to finish the reaction; firstly, washing with acetone, then replacing acetone in the beads with acetone aqueous solutions with different proportions, gradually adjusting the acetone aqueous solutions to 25 vol%, 50 vol% and 75 vol%, and finally washing with water to obtain phenyl-containing agarose beads;
S32, mixed-mode medium synthesis: the agarose beads containing phenyl are crosslinked with 2, 3-epoxypropyltrimethylammonium chloride, 100ml of the agarose beads containing phenyl are poured into a reactor, 100ml of 50 wt% NaOH is added, the mixture is fully mixed, GTA is added, the mixture is continuously stirred, the reaction is stopped after 3 hours of reaction at the temperature of 50 ℃, ethanol and deionized water are alternately used for washing for 3 times, and finally the product is obtained and stored in 20 wt% ethanol.
2. The method of claim 1, wherein in the step of synthesizing phenyl glycidyl ether in step S1: mixing phenol, epoxy chloropropane, PEG and trichloromethane, heating to 35-45 ℃, and reacting for 8-12 h; and after the reaction is finished, filtering, washing with water, distilling under reduced pressure, and collecting a product of a fraction at 93-95 ℃ to obtain the phenyl glycidyl ether.
3. The method of claim 2, wherein the anion exchange medium is selected from the group consisting of: the molar ratio of the phenol to the epichlorohydrin is 1: 2; the volume ratio of the PEG to the chloroform solvent is 1: 10.
4. The method of claim 1, wherein in the step of synthesizing the ligand 2, 3-epoxypropyltrimethylammonium chloride of step S2: putting a certain amount of epoxy chloropropane into a reactor, cooling to 0-5 ℃, continuously stirring, adding a certain amount of trimethylamine hydrochloride, and reacting for 3-5 hours at normal temperature; and after the reaction is finished, filtering, washing for 3-5 times by using acetone, and drying in vacuum to obtain the GTA white solid.
5. The method of claim 4, wherein the anion exchange medium is selected from the group consisting of: the molar ratio of trimethylamine hydrochloride to epichlorohydrin is 1: (4-6).
6. the method of claim 5 for preparing a mixed anion exchange media, wherein: the molar ratio of trimethylamine hydrochloride to epichlorohydrin is 1: 5.
7. A mixed anion exchange medium prepared according to the preparation method of any one of claims 1 to 6.
8. a mixed anion exchange medium has a structure shown in formula (I),
The above-mentionedAgarose beads are indicated.
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