CN108276526A - A kind of high carrying capacity large aperture polymer cation displacement chromatography medium and its preparation - Google Patents
A kind of high carrying capacity large aperture polymer cation displacement chromatography medium and its preparation Download PDFInfo
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- 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
- C08F265/00—Macromolecular compounds obtained by polymerising monomers on to polymers of unsaturated monocarboxylic acids or derivatives thereof as defined in group C08F20/00
- C08F265/04—Macromolecular compounds obtained by polymerising monomers on to polymers of unsaturated monocarboxylic acids or derivatives thereof as defined in group C08F20/00 on to polymers of esters
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- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J39/00—Cation exchange; Use of material as cation exchangers; Treatment of material for improving the cation exchange properties
- B01J39/08—Use of material as cation exchangers; Treatment of material for improving the cation exchange properties
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- C08F257/00—Macromolecular compounds obtained by polymerising monomers on to polymers of aromatic monomers as defined in group C08F12/00
- C08F257/02—Macromolecular compounds obtained by polymerising monomers on to polymers of aromatic monomers as defined in group C08F12/00 on to polymers of styrene or alkyl-substituted styrenes
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- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F267/00—Macromolecular compounds obtained by polymerising monomers on to polymers of unsaturated polycarboxylic acids or derivatives thereof as defined in group C08F22/00
- C08F267/06—Macromolecular compounds obtained by polymerising monomers on to polymers of unsaturated polycarboxylic acids or derivatives thereof as defined in group C08F22/00 on to polymers of esters
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- C08F2438/01—Atom Transfer Radical Polymerization [ATRP] or reverse ATRP
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Abstract
The invention discloses belong to polymer chromatography medium to prepare and a kind of high carrying capacity large aperture polymer cation displacement chromatography medium of applied technical field and its preparation.The cation-exchange chromatography medium is using polyacrylate or polystyrene type microballoon as matrix, microsphere surface grafting vinyl function monomer.Preparation method includes：Polyacrylate or polystyrene type microballoon are subjected to swelling treatment in vinyl function monomer solution；Atom transfer radical polymerization catalyst-ligand-system is added into the microballoon after swelling treatment, carries out graft polymerization reaction, after polymerisation, removes unreacted monomer, vacuum drying obtains cation-exchange chromatography medium.The cation-exchange chromatography medium ionic exchange capacity of preparation is high, protein binding carrying capacity is high, operation flow velocity is fast, is suitble to the fast separating and purifying of large biological molecule.
Description
Technical field
The invention belongs to the preparation of polymer chromatography medium and applied technical fields, and in particular to a kind of high carrying capacity large aperture is poly-
Close object cation-exchange chromatography medium and its preparation.
Background technology
With the rapid development of medicine biological technique, bio-pharmaceuticals production scale is growing, pure to separation downstream
Change proposes new challenge, and the purification process developed rapidly and efficiently is the task of top priority.Wherein separating medium is separating and purifying technology
Core, currently used separating medium is mostly using polysaccharide as matrix, and since quality itself is soft, (pressure resistance is less than such medium
0.3MPa), it the features such as aperture size small (30-50nm), is subject to certain restrictions in high-throughput isolating and purifying is applied.In order to
It realizes high-throughput separation, becomes the only selection of people by the macropore chromatography media of matrix of polymer.Macroporous polymer is micro-
Ball has high mechanical strength (pressure resistance is up to 10MPa), range of aperture size big (20-500nm), therefore can be in higher operation stream
It is detached under speed, substantially reduces the operating time.But the increase in the aperture with such polymer microballoon, specific surface area go out
Corresponding decline is showed, specific surface area is often in 10m2/ g is hereinafter, cause the protein load of unit volume medium to reduce, therefore be somebody's turn to do
Class macroporous matrix is difficult to take into account high flow rate and high carrying capacity in actual separation purifying, finally cannot achieve real high throughput.
To solve the above-mentioned problems, generally use increases the method for material surface functional group quantity, such as open source literature
(Journal of Chromatography A, 2014,1343,109-118) is reported with containing multiple functional groups (amino)
Macromolecular polyethyleneimine (PEI) modify macropore polyacrylate microballoon, by the epoxy group of amino and microsphere surface into
Then row coupling reaction is again branched it using the excessive crosslinking agent with bis-epoxy, after branched, then using excessive
PEI be bonded, in this way, by 3 steps react, finally improve the function radix amount of microsphere surface, improve the egg of medium
Bai Zailiang.Reactions steps of this method is more, and since the reaction of every step is difficult to quantitative control, therefore needs that excessive reaction reagent is added,
A certain amount of waste is caused, manufacturing cost is increased.Therefore, exploitation one kind is easy to operate, can accurately control material surface work(
Can the method for radical amount promote the protein load of medium, be the technology being badly in need of in bio-chemistry separation purification art.
Invention content
The purpose of the present invention is to propose to a kind of high carrying capacity large aperture polymer cation displacement chromatography medium and its preparations.Tool
Body technique scheme is as follows：
A kind of high carrying capacity large aperture polymer cation displacement chromatography medium, it is micro- with polyacrylate or polystyrene type
Ball is matrix, microsphere surface grafting vinyl function monomer.
The polyacrylate or modified product, derivative that polystyrene type microballoon is polyacrylate or polystyrene
Product or graft copolymer.
The polyacrylate or polystyrene type microballoon are glycidyl methacrylate and divinylbenzene
Copolymerization microsphere, the copolymerization microsphere of glycidyl methacrylate and ethylene glycol dimethacrylate, styrene and divinyl
The copolymerization microsphere of base benzene, the copolymerization microsphere of styrene and ethylene glycol dimethacrylate, ethylene glycol dimethacrylate
The autohemagglutination microballoon of autohemagglutination microballoon or divinylbenzene.
The polyacrylate or polystyrene type microballoon are prepared using atom transfer radical polymerization method, described micro-
The aperture size of ball is 100-3000nm, grain size 20-200um, specific surface area 5-200m2/ g, operating pressure reach
10MPa。
The vinyl function monomer be acrylic acid, 2- acrylamidos -2- methyl-1s-propane sulfonic acid, methacrylic acid and
One or more of methylpropene sodium sulfonate.
The preparation method of above-mentioned high throughput large aperture polymer cation displacement chromatography medium, includes the following steps：
(1) polyacrylate or polystyrene type microballoon are subjected to swelling treatment in vinyl function monomer solution；
(2) atom transferred free radical being made of inorganic halides and multiple tooth amine is added into the microballoon after swelling treatment
Polymerization catalyst ligand system, is stirred, and carries out graft polymerization reaction, after polymerisation, remove unreacted monomer and
Other impurities, vacuum drying, obtain cation-exchange chromatography medium.
The operation of swelling treatment described in step (1) is：Under air-proof condition, 80~120rpm, 0.5~2h of mechanical agitation, the phase
Between be filled with high pure nitrogen and protected.
Vinyl function monomer described in step (1) is acrylic acid, 2- acrylamidos -2- methyl-1s-propane sulfonic acid, methyl
One or more of acrylic acid and methylpropene sodium sulfonate；
The vinyl function monomer solution solvent for use is dioxane, methanol, dimethyl sulfoxide (DMSO), dimethylformamide
With it is more than one or two kinds of in deionized water；
The volumetric usage of the solvent is 3~10 times of the microspheres quality, i.e. microspheres quality：Solvent volume=1:3~
10(g/mL)。
The reaction temperature of polymerisation described in step (2) be 30 DEG C~80 DEG C, the reaction time be 6~for 24 hours.
Inorganic halides described in step (2) are one or both of cuprous bromide, stannous chloride, and addition is poly-
1/70~1/20 times of esters of acrylic acid or polystyrene type microspheres quality；The multiple tooth amine 2,2- bipyridines, tetramethyl
Ethylenediamine, N, N, N, ' N, " N, " ' in-five methyl diethylentriamine and 1,1,4,7,10,10 '-hexamethyl triens
One or more, addition is 3 times of inorganic halide amount of substance.
It is filtered and is washed with water by decompression in step (2) to remove unreacted monomer and other impurities；The vacuum is dry
Dry temperature is 40 DEG C~60 DEG C, and the time is 20~28h.
Beneficial effects of the present invention are：
(1) present invention is in order to promote the protein load of polyacrylate or polystyrene type micro-sphere material, using atom
The method of transferring free-radical polymerization (ATRP) causes vinyl function monomer in polyacrylate or polystyrene type microballoon table
Face and internal channel surfaces are graft-polymerized；Polyacrylate or the micro- atomic radicals that may be used of polystyrene type polymerize
Prepared by method, microsphere surface remains the initiator group (halides) of ATRP, such group can continue to cause vinyl monomer
It is polymerize；The ion-exchange group quantity of microsphere surface after grafting increases, and ion exchange capacity improves, and it is micro- to increase macropore
The protein binding carrying capacity of ball.The range of aperture size of polyacrylate or polystyrene type micro-sphere material is 100-3000nm,
Grain size is 20-200um, specific surface area ranging from 5-200m2/ g has higher mechanical strength and fluid mass-transfer performance；
Ion exchange capacity is 1.2-4.0mmol/g after material surface and internal channel surfaces grafting vinyl function monomer, and albumen carries
Amount ranging from 48-234mg/mL media have the characteristics that ion exchange capacity is high, protein binding carrying capacity is high, operation flow velocity is fast, make
It is suitably applied high-throughput isolation purifying biological macromolecular for cation-exchange chromatography medium, is applied to chromatographic isolation field.
(2) for the vinyl function monomer used for small molecule monomer, reaction steric hindrance is small, can diffuse to porous material completely
Duct inside, solve the problems, such as that material duct built-in function group caused by previous macromolecular steric hindrance is non-uniform；Simultaneously
ATRP is a kind of activity/controllable polymerization mode, the grafting chain length of material surface can be effectively controlled, so as to accurately control material
Expect the functional group quantity of surface and internal channel surfaces.
Description of the drawings
Fig. 1 is that large aperture polymer microballoon surface graft copolymerization reaction prepares the signal of cation-exchange chromatography medium process
Figure.
Fig. 2 be the gained of embodiment 1 polymer microballoon before modified after electron scanning micrograph, wherein A is to be modified
Preceding microballoon, B are modified microballoon.
Fig. 3 is the frontal analysis chromatogram of cation exchange medium prepared by embodiment 1.
Fig. 4 is the frontal analysis chromatogram of cation exchange medium prepared by embodiment 2.
Fig. 5 is the frontal analysis chromatogram of cation exchange medium prepared by embodiment 3.
Fig. 6 is the frontal analysis chromatogram of cation exchange medium prepared by embodiment 4.
Fig. 7 is the frontal analysis chromatogram of cation exchange medium prepared by embodiment 5.
Fig. 8 is the frontal analysis chromatogram of cation exchange medium prepared by embodiment 6.
Specific implementation mode
Following embodiment facilitates a better understanding of the present invention.
Embodiment 1：High carrying capacity large aperture PGMA-EDMA cation-exchange chromatography media
Macroporous polymer microsphere surface graft copolymerization prepares cation-exchange chromatography medium process schematic such as Fig. 1.
The preparation method of high carrying capacity large aperture PGMA-EDMA cation-exchange chromatography media, includes the following steps：
(1) large aperture poly (glycidyl methacrylate)-ethylene glycol dimethacrylate (PGMA-EDMA) microballoon
(average pore size 128nm) swelling treatment in acrylic acid solution
It accurately weighs PGMA-EDMA microballoons 1.0g to be put into three mouthfuls of reaction bulbs of 50mL, the diformazan of acrylic acid is then added
It is swollen 2.0h under base sulfoxide solution 3.0mL, 100rpm mechanical agitation, being filled with high pure nitrogen during swelling treatment is protected.
(2) PGMA-EDMA microsphere surfaces graft polymerization acrylic acid
Weigh 0.014g CuCl (1/70 microspheres quality) and 0.042g 2,2- bipyridines (Bpy) (3 times of CuCl mass)
It is added in the microballoon after step (1) swelling treatment, keeps 30 DEG C of temperature, keep reaction 6h at this temperature, after reaction
While hot, decompression suction filtration is carried out with G3 sand core funnels, while is washed with 500mL deionized waters colourless to microballoon.It is micro- after grafting
Ball surface and channel surfaces have polyacrylic acid long-chain molecule, since the polymer lateral chain carries a large amount of-COOH group,
It can be directly used as weak cation exchange chromatography media.Fig. 2 be the present embodiment obtained by polymer microballoon before modified after electronics
Flying-spot microscope photo, wherein A and B be respectively before modified after polymer microballoon.Fig. 3 is cation prepared by the present embodiment
The frontal analysis chromatogram of exchange media.
The grafting amount of gravimetric detemination microsphere surface polyacrylic acid is 108mg/g, and measuring its ion exchange capacity is
1.20mmol/g, it is 180mg/mL media that lysozyme, which adsorbs carrying capacity,.
The amount of the graft polymerization of every gram of microballoon is calculated with gravimetric method, calculation formula is as follows：
Ion exchange capacity measures：The modified microballoons of 1.0g are loaded in the glass chromatography column that internal diameter is 10mm, are divided such as
Under several steps carry out ion exchange capacity measurement：
1) it is washed with the aqueous hydrochloric acid solution 20mL of 1.0mol/L, medium is transformed into Hydrogen；2) it is washed with deionized water
It is neutrality to wash to outflow cleaning solution；3) sodium hydroxide solution for accurately measuring 50mL 0.1mol/L is added in chromatographic column, is allowed to certainly
So drips and collect the lye；4) sodium-chloride water solution of 20mL 1.0mol/L is added, gives free rein to drip, and collects with 4)
In lye merge；5) two drop methyl orange indicators are added into the collection liquid in 4), are titrated with the hydrochloric acid solution of 0.1mol/L
Merge collection liquid, the ion exchange capacity of calculation medium.
The measurement of matrix protein combination carrying capacity：Appropriate cation-exchange chromatography medium is taken to be packed intoChromatographic column
In (medium volume 1.0mL), the chromatographic column filled is connected on liquid chromatograph, chromatographic condition is as follows：
The lysozyme soln of sample concentration 2mg/mL；Pump head loading；Flow velocity 1mL/min；Mobile phase A pH=7.0,50mM
Phosphate buffer solution；Mobile phase B 1mol/L NaCl pH=7.0 50mM phosphate buffer solutions；Color is balanced with mobile phase A first
10 column volumes (CV) of column are composed, then pump head loading protein solution to 100% flows through, then is carried out to adhesion protein with Mobile phase B
Elution, records corresponding chromatogram and flows through value according to flow through curve calculation medium 5%, obtain the dynamic protein binding of medium
Amount.
5% penetration volume v5% is recorded, the dynamic adsorbance of lysozyme is calculated, formula is as follows：
F tests flow velocity, mL/min；C, lysozyme sample concentration, mg/mL；V, medium volume, mL.
Embodiment 2：High carrying capacity large aperture PEDMA cation-exchange chromatography media
The preparation method of high carrying capacity large aperture PEDMA cation-exchange chromatography media, includes the following steps：
(1) macropore PEDMA microballoons (average pore size 687nm) are molten in 2- acrylamidos -2- methyl-1s-propanesulfonic acid solutions
Swollen processing
It accurately weighs PEDMA microballoons 1.0g to be put into three mouthfuls of reaction bulbs of 50mL, 2- acrylamido -2- first is then added
It is swollen 1.0h under dimethyl formamide solution 5.0mL, the 100rpm mechanical agitation of base -1- propane sulfonic acid, is filled with during swelling treatment
High pure nitrogen is protected.
(2) PEDMA microsphere surfaces graft polymerization 2- acrylamidos -2- methyl-1s-propane sulfonic acid
Weigh 0.049g CuCl (1/20 microspheres quality) and 0.147g tetramethylethylenediamines (TMEDA) (3 times of CuCl mass)
It is added in the microballoon after step (1) swelling treatment, keeps temperature 60 C, keep reaction for 24 hours at this temperature, after reaction
While hot, decompression suction filtration is carried out with G3 sand core funnels, while is washed with 500mL deionized waters colourless to microballoon.It is micro- after grafting
Ball surface and channel surfaces have poly- 2- acrylamidos -2- methyl-1s-propane sulfonic acid long-chain molecule, due to the polymer side chain belt
There is a large amount of sulfonic acid group, therefore strong cation exchange chromatography media can be directly used as.Fig. 4 be the present embodiment prepared by sun from
The frontal analysis chromatogram of sub- exchange media.
The grafting amount of the poly- 2- acrylamidos -2- methyl-1s-propane sulfonic acid of gravimetric detemination microsphere surface is 416mg/g, is surveyed
Its fixed ion exchange capacity is 1.9mmol/g, and it is 174mg/mL media that lysozyme, which adsorbs carrying capacity,.Grafting amount, ion exchange capacity
The assay method of carrying capacity is adsorbed with embodiment 1 with lysozyme.
Embodiment 3：High carrying capacity large aperture PSt-EDMA cation-exchange chromatography media
The preparation method of high carrying capacity large aperture PSt-EDMA cation-exchange chromatography media, includes the following steps：
(1) macropore PSt-EDMA microballoons (average pore size 2100nm) pre-process in methacrylic acid solution
It accurately weighs PSt-EDMA microballoons 1.0g to be put into three mouthfuls of reaction bulbs of 50mL, is then added the two of methacrylic acid
It is swollen 0.5h under six ring solution 10mL, 100rpm mechanical agitation of oxygen, being filled with high pure nitrogen during swelling treatment is protected.
(2) macropore PSt-EDMA microsphere surfaces graft polymerization methacrylic acid
0.016g CuCl (1/60 microspheres quality) and 0.048g N, N, N are weighed, ' N, " N, " '-pentamethyl diethylidene three
Amine (PMDETA) (3 times of CuCl mass) is added in the microballoon after step (1) swelling treatment, 40 DEG C of temperature is kept, in this temperature
Lower holding reaction 10h carries out decompression suction filtration, while being carried out with 500mL deionized waters with G3 sand core funnels after reaction while hot
Washing is colourless to microballoon.Fig. 5 is the frontal analysis chromatogram of cation exchange medium prepared by the present embodiment.
Microsphere surface and channel surfaces have polymethylacrylic acid long-chain molecule after grafting, since the polymer lateral chain carries
Largely-COOH group, therefore weak cation exchange chromatography media can be directly used as.The poly- methyl of gravimetric detemination microsphere surface
The grafting amount of acrylic acid is 367mg/g, and it is 4.0mmol/g to measure its ion exchange capacity, and it is 228mg/ that lysozyme, which adsorbs carrying capacity,
ML media.The assay method of grafting amount, ion exchange capacity and lysozyme absorption carrying capacity is the same as embodiment 1.
Embodiment 4：High carrying capacity large aperture PSt-DVB cation-exchange chromatography media
The preparation method of high carrying capacity large aperture PSt-DVB cation-exchange chromatography media, includes the following steps：
(1) macropore PSt-DVB microballoons (average pore size 3010nm) pre-process in methacrylic acid solution
It accurately weighs PSt-DVB microballoons 1.0g to be put into three mouthfuls of reaction bulbs of 50mL, the first of methacrylic acid is then added
It is swollen 1.0h under alcoholic solution 10mL, 100rpm mechanical agitation, being filled with high pure nitrogen during swelling treatment is protected.
(2) macropore PSt-DVB microsphere surfaces graft polymerization methacrylic acid
Weigh 0.019g CuCl (1/50 microspheres quality) and 0.058g 1,1,4,7,10,10 '-hexamethyls triethylene four
Amine (HMETETA) (3 times of CuCl mass) is added in the microballoon after step (1) swelling treatment, temperature 50 C is kept, in this temperature
Lower holding reaction 15h carries out decompression suction filtration, while being carried out with 500mL deionized waters with G3 sand core funnels after reaction while hot
Washing is colourless to microballoon.Microsphere surface and channel surfaces have polymethylacrylic acid long-chain molecule after grafting, due to the polymer
Side chain can be directly used as weak cation exchange chromatography media with a large amount of-COOH group.Fig. 6 is the present embodiment institute
Prepare the frontal analysis chromatogram of cation exchange medium.
The grafting amount of gravimetric detemination microsphere surface polymethylacrylic acid is 245mg/g, and measuring its ion exchange capacity is
2.2mmol/g, it is 48mg/mL media that lysozyme, which adsorbs carrying capacity,.The survey of grafting amount, ion exchange capacity and lysozyme absorption carrying capacity
Method is determined with embodiment 1.
Embodiment 5：High carrying capacity large aperture PDVB cation-exchange chromatography media
The preparation method of high carrying capacity large aperture PDVB cation-exchange chromatography media, includes the following steps：
(1) macropore PDVB microballoons (average pore size 1203nm) pre-process in acrylic acid solution
It accurately weighs PDVB microballoons 1.0g to be put into three mouthfuls of reaction bulbs of 50mL, the dimethyl sulfoxide (DMSO) of acrylic acid is then added
It is swollen 1.5h under solution 10mL, 100rpm mechanical agitation, being filled with high pure nitrogen during swelling treatment is protected.
(2) macropore PDVB microsphere surfaces graft polymerization acrylic acid
Weigh 0.019g CuBr (1/50 microspheres quality) and 0.058g 1,1,4,7,10,10 '-hexamethyls triethylene four
Amine (HMETETA) (3 times of CuBr mass) is added in the microballoon after step (1) swelling treatment, 30 DEG C of temperature is kept, in this temperature
Lower holding reaction 20h carries out decompression suction filtration, while being carried out with 500mL deionized waters with G3 sand core funnels after reaction while hot
Washing is colourless to microballoon.Microsphere surface and channel surfaces have polyacrylic acid long-chain molecule after grafting, due to the polymer lateral chain
With a large amount of-COOH group, therefore weak cation exchange chromatography media can be directly used as.Fig. 7 is prepared by the present embodiment
The frontal analysis chromatogram of cation exchange medium.
The grafting amount of gravimetric detemination microsphere surface polymethylacrylic acid is 256mg/g, and measuring its ion exchange capacity is
2.9mmol/g, it is 87mg/mL media that lysozyme, which adsorbs carrying capacity,.The survey of grafting amount, ion exchange capacity and lysozyme absorption carrying capacity
Method is determined with embodiment 1.
Embodiment 6：High carrying capacity large aperture PGMA-DVB cation-exchange chromatography media
The preparation method of high carrying capacity large aperture PGMA-DVB cation-exchange chromatography media, includes the following steps：
(1) macropore PGMA-DVB microballoons (average pore size 978nm) pre-process in methylpropene sodium sulfonate solution
It accurately weighs PDVB microballoons 1.0g to be put into three mouthfuls of reaction bulbs of 50mL, the water of methylpropene sodium sulfonate is then added
It is swollen 0.5h under solution 6mL, 100rpm mechanical agitation, being filled with high pure nitrogen during swelling treatment is protected.
(2) macropore PGMA-DVB microballoons (average pore size 978nm) surface grafting polymerization methylpropene sodium sulfonate
Weigh 0.024g CuBr (1/40 microspheres quality) and 0.071g 1,1,4,7,10,10 '-hexamethyls triethylene four
Amine (HMETETA) (3 times of CuBr mass) is added in the microballoon after step (1) swelling treatment, 40 DEG C of temperature is kept, in this temperature
Lower holding reaction for 24 hours, after reaction while hot, carries out decompression suction filtration, while being carried out with 500mL deionized waters with G3 sand core funnels
Washing is colourless to microballoon.Microsphere surface and channel surfaces have polymethyl sodium sulfonate long-chain molecule after grafting, since this is poly-
It closes object side chain and carries a large amount of sodium group, therefore strong cation exchange chromatography media can be directly used as.Fig. 8 is this implementation
The frontal analysis chromatogram of cation exchange medium prepared by example.
The grafting amount of gravimetric detemination microsphere surface polymethyl sodium sulfonate is 356mg/g, measures its ion exchange appearance
Amount is 2.0mmol/g, and it is 234mg/g media that lysozyme, which adsorbs carrying capacity,.Grafting amount, ion exchange capacity and lysozyme adsorb carrying capacity
Assay method with embodiment 1.
Claims (11)
1. a kind of high carrying capacity large aperture polymer cation displacement chromatography medium, which is characterized in that the cation-exchange chromatography
Medium is using polyacrylate or polystyrene type microballoon as matrix, microsphere surface grafting vinyl function monomer.
2. polymer cation displacement chromatography medium according to claim 1, which is characterized in that the polyacrylate
Or modified product, derivative products or the graft copolymer that polystyrene type microballoon is polyacrylate or polystyrene.
3. polymer cation displacement chromatography medium according to claim 2, which is characterized in that the polyacrylate
Or polystyrene type microballoon is sweet for copolymerization microsphere, the Glycidyl methacrylate of glycidyl methacrylate and divinylbenzene
The copolymerization microsphere of grease and ethylene glycol dimethacrylate, the copolymerization microsphere of styrene and divinylbenzene, styrene and second
Copolymerization microsphere, the autohemagglutination microballoon of ethylene glycol dimethacrylate or the autohemagglutination of divinylbenzene of diol dimethacrylate
Microballoon.
4. polymer cation displacement chromatography medium according to claim 2, which is characterized in that the polyacrylate
Or polystyrene type microballoon is prepared using atom transfer radical polymerization method, the aperture size of the microballoon is 100-
3000nm, grain size 20-200um, specific surface area 5-200m2/ g, operating pressure reach 10MPa.
5. polymer cation displacement chromatography medium according to claim 1, which is characterized in that the vinyl function list
Body be acrylic acid, one kind in 2- acrylamidos -2- methyl-1s-propane sulfonic acid, methacrylic acid and methylpropene sodium sulfonate or
It is two or more.
6. the preparation method of any one of the claim 1-5 high-throughput large aperture polymer cation displacement chromatography media,
It is characterized in that, includes the following steps：
(1) polyacrylate or polystyrene type microballoon are subjected to swelling treatment in vinyl function monomer solution；
(2) atom transfer radical polymerization being made of inorganic halides and multiple tooth amine is added into the microballoon after swelling treatment
Catalyst-ligand-system is stirred, and carries out graft polymerization reaction, after polymerisation, remove unreacted monomer and other
Impurity, vacuum drying, obtains cation-exchange chromatography medium.
7. preparation method according to claim 6, which is characterized in that the operation of swelling treatment described in step (1) is：It is close
Under the conditions of envelope, during which 80~120rpm, 0.5~2h of mechanical agitation are filled with high pure nitrogen and are protected.
8. preparation method according to claim 6, which is characterized in that vinyl function monomer described in step (1) is third
One or both of olefin(e) acid, 2- acrylamidos -2- methyl-1s-propane sulfonic acid, methacrylic acid and methylpropene sodium sulfonate with
On；
The vinyl function monomer solution solvent for use is dioxane, methanol, dimethyl sulfoxide (DMSO), dimethylformamide and goes
It is more than one or two kinds of in ionized water；
The volumetric usage of the solvent is 3~10 times of the microspheres quality.
9. preparation method according to claim 6, which is characterized in that the reaction temperature of polymerisation described in step (2)
Be 30 DEG C~80 DEG C, the reaction time be 6~for 24 hours.
10. preparation method according to claim 6, which is characterized in that inorganic halides described in step (2) are protobromide
One or both of copper, stannous chloride, addition be polyacrylate or polystyrene type microspheres quality 1/70~
1/20 times；The multiple tooth amine 2,2- bipyridines, tetramethylethylenediamine, N, N, N, ' N, " N, " '-pentamethyl diethylidene three
Amine and 1, Isosorbide-5-Nitrae, one or more of 7,10,10 '-hexamethyl triens, addition are inorganic halides
3 times of quality.
11. preparation method according to claim 6, which is characterized in that filter and be washed with water by depressurizing in step (2)
To remove unreacted monomer and other impurities；The vacuum drying temperature is 40 DEG C~60 DEG C, and the time is 20~28h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810078358.4A CN108276526B (en) | 2018-01-26 | 2018-01-26 | High-load large-aperture polymer cation exchange chromatography medium and preparation thereof |
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| CN109265613A (en) * | 2018-08-20 | 2019-01-25 | 南通乐道环保技术有限公司 | A kind of functional polystyrene microballoon and its preparation method and application |
| CN113683727A (en) * | 2021-05-30 | 2021-11-23 | 苏州星谱生物科技有限公司 | Cation exchange medium and preparation method thereof |
| CN114014913A (en) * | 2022-01-10 | 2022-02-08 | 浙江湃肽生物有限公司南京分公司 | Purification method of triptorelin acetate |
| CN114989359A (en) * | 2022-06-22 | 2022-09-02 | 苏州纳微科技股份有限公司 | Cation exchange chromatography medium and preparation method thereof |
| CN115850792A (en) * | 2022-09-30 | 2023-03-28 | 北京石油化工学院 | Preparation method of high-load anion exchange chromatography medium |
| CN116120561A (en) * | 2022-12-26 | 2023-05-16 | 宁波争光树脂有限公司 | Phosphoric acid resin and preparation method thereof |
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| CN115850792B (en) * | 2022-09-30 | 2024-01-30 | 北京石油化工学院 | Preparation method of high-loading anion exchange chromatography medium |
| CN116120561A (en) * | 2022-12-26 | 2023-05-16 | 宁波争光树脂有限公司 | Phosphoric acid resin and preparation method thereof |
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