CN108948385A - A kind of synthetic method of high adsorption capacity strong acid agar base chromatography media - Google Patents
A kind of synthetic method of high adsorption capacity strong acid agar base chromatography media Download PDFInfo
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Abstract
The present invention relates to the bio-separation of bioengineering and cell and its pharmaceutical carrier fields, and in particular to a kind of synthetic method of high adsorption capacity strong acid agar base chromatography media.The present invention is using agarose solution as water phase, toluene is oily phase, carries out functionalization and cross-linking reaction to the forming process of agarose microbeads using 3- chlorine-2-hydroxyl propanesulfonate and epoxychloropropane, strong acid functionalization, cross-linking reaction and one step of balling-up carry out, it is finally post-processed, obtains product.This invention simplifies preparation processes, save the preparation cost, and the strong acid agar base chromatography media mechanical strength of acquisition is good, function base is evenly distributed and adsorbance is high.
Description
Technical field
The present invention relates to the bio-separation of bioengineering and cell and its pharmaceutical carrier fields, and in particular to a kind of high absorption
Measure the synthetic method of strong acid agar base chromatography media.
Background technique
In recent years, upstream, cell culture and expression technology are constantly mature, production scale continuous enlargement, biotechnology it is swift and violent
Develop and bring huge pressure to downstream separation purifying, it is desirable that downstream has bigger treating capacity and higher production efficiency.
Ion-exchange chromatography (ion-exchange chromato-graphy, IEC) is with Ion Exchange Medium for fixation
Phase, according in mobile phase between ionic sample and medium charged group electrostatic interaction strong and weak difference, realize point of solute
From.The technology is widely applied in bioengineering due to the characteristics of its is easy to operate, applied widely, selectivity is high, easy regeneration
In each step of the separation of large biological molecule (such as protein, polypeptide, nucleic acid, polynucleotide), medium purifying and purification
In, it is one of the most effectual way of biological downstream process.
The development of ion-exchange chromatography depends on ion-exchange chromatography media.Select function admirable, low-cost
Separating medium is the key that chromatographic technique.The separating property of Ion Exchange Medium is mainly by the shadow of two factors of aglucon and matrix
It rings.Wherein, aglucon provides tradable ion, and matrix provides skeleton.Ago-Gel is high with its good hydrophilic property, biocompatibility
The characteristics of be used widely in bio-separation engineering field.
For highly acidic cation agarose chromatography medium, general preparation step is divided into three steps: (1) preparing agar
Sugared microballoon；It (2) will be micro-sphere crosslinked；(3) the bonded functional group on crosslinked microsphere.The not Ago-Gel Jing Guo any crosslinking
The pressure that microballoon is born is limited, and flow velocity is slower；The agarose gel microsphere being crosslinked, mechanical strength are remarkably reinforced, but due to
Common cross-linking agent is mostly oil-soluble, is difficult in the aperture for uniformly entering the agarose gel microsphere that moisture is 95% or so
Even crosslinking, therefore micro-sphere structure and surface-crosslinked degree have difference to influence its mechanical performance；Higher crosslinking degree simultaneously, can limit
Diffusion of the activating reagent processed into ball, cause active group and aglucon be unevenly distributed and the decline of density, and then influence
Product separating property.
Summary of the invention
The strong acid fine jade that the object of the present invention is to provide a kind of processing machinery intensity is good, function base is evenly distributed and adsorbance is high
The synthetic method of aliphatic radical chromatography media.
The present invention is achieved by the following technical solutions: using agarose solution as water phase, toluene is oily phase, using 3-
Chlorine-2-hydroxyl propanesulfonate and epoxychloropropane carry out functionalization and cross-linking reaction to the forming process of agarose microbeads, finally
It is post-processed, obtains better mechanical property, the strong acid agarose gel microsphere that function base is evenly distributed.This method mainly includes
The processes such as water phase preparation, the preparation of suspension, strong acid functionalization and cross-linking reaction, solidification and post-processing.
The specific steps are that:
A) prepared by water phase
The agarose solution that mass concentration is 4-8% is prepared, is heated to 90 DEG C or more, agarose all dissolves, and is added
3- chlorine-2-hydroxyl propanesulfonate solid is stirred to dissolution, obtains water phase；
B) prepared by suspension
Water phase is poured into 70-80 DEG C of Toluene oil phase, oil is mutually 6-8:1 with the volume ratio of water phase, obtains suspension；
C) strong acid functionalization and cross-linking reaction
Epoxychloropropane is added into step b) suspension, and the NaOH solution that mass concentration is 50% is added dropwise, and in 1h
Inside it is added dropwise；70-75 DEG C is maintained the temperature at, 3-6h is reacted, obtains cross-linking reaction product；
D) solidify
By step c) cross-linking reaction product, is condensed, be cooled to 10 DEG C or less；Partial size is prepared after being cleaned multiple times is
The cured product of 100-300 mesh；
E) it post-processes
The cured product prepared in step d) is weighed, is added in ethanol solution, is then added NaOH solution, 40-60 DEG C
Lower constant temperature stirs 8-16h, obtains product, i.e. strong acid agarose gel microsphere.
Wherein, the mass ratio of 3- chlorine-2-hydroxyl propanesulfonate and agarose described in step a) is 1.5-2.5:1；Institute
Potassium chloride is added in the agarose solution stated, and the mass concentration of potassium chloride is 10%.KCl ion reduces agarose charge
The charge and hydration of molecule discharge Free water, so that agarose viscosity declines.It is molten that the decline of viscosity is more favorable for agarose
Liquid is evenly dispersed in oily phase, is conducive to the hydroxyl on later period 3- chlorine-2-hydroxyl propanesulfonate and epoxychloropropane and agarose
Base contact carries out functionalization and cross-linking reaction, guarantees the uniformity and higher mechanical strength of the distribution of ball function base.
In the present invention, 3- chlorine-2-hydroxyl propanesulfonate provide strong acid group-sulfonic group, 3- chlorine-2-hydroxyl propanesulfonate with
The group after hydroxyl reaction on agarose is-CH2CH(OH)CH2SO3, there are a hydroxyls on sulfonic group adjacent bit C, increase
The hydrophily of the group is added.Hydroxyl can form hydrogen bond with albumen, and adhesion protein is stronger, and when elution can make hydrogen bond
It is more sensitive to the variation of salinity, it is easy to retain protein active and avoids being denaturalized.The amount of 3- chlorine-2-hydroxyl propanesulfonate is excessive,
Be relative to the hydroxyl on agarose it is excessive, cannot be reacted with hydroxyl, while will affect the viscosity of agarose and then leading to fine jade
The granularity of lipolysaccharide becomes smaller, and is unfavorable for the control of particle size uniformity, therefore, the quality of 3- chlorine-2-hydroxyl propanesulfonate and agarose
Than for 1.5-2.5:1.
In addition, the present invention is that 3- chlorine-2-hydroxyl propanesulfonate solid is added in the aqueous solution of agarose first, with fine jade
The aqueous solution of lipolysaccharide is poured into Toluene oil phase again after being sufficiently mixed.3- chlorine-2-hydroxyl propanesulfonate solid is added to
In the aqueous solution of agarose, it ensure that strong acid group in the uniformity of agar sugar ball inner part cloth, solves traditional strong acid ion
Group is in chromatography media the problem of distributing inhomogeneity.Water phase is added in oily phase, also ensures the uniformity of mixing.If
The aqueous solution of agarose is first poured into oily phase by reversed order, and agarose viscosity can be made to increase, add the chloro- 2- hydroxyl of 3- at this time
Base propanesulfonate is then difficult uniformly to mix with agarose again；Epoxychloropropane is added at this time, on epoxychloropropane and agarose
Hydroxyl carry out cross-linking reaction, also will affect the uniform mixability of 3- chlorine-2-hydroxyl propanesulfonate and agarose.
The polyvinyl acetate and oleic acid contained in Toluene oil phase in step b), quality are respectively the 1- of toluene quality
2% and 0.8-1.2%.The application of polyvinyl acetate and oleic acid ensure that the product mechanical strength finally obtained is good.
The mass ratio of epoxychloropropane described in step c) and agarose is 3:1；The body of the 50%NaOH solution of dropwise addition
Product is 1.5-3 times of volume of epoxychloropropane.In the step, the additional amount of sodium hydroxide solution is excessive, 70 DEG C at a temperature of
Aqueous slkali is damaged to a certain extent effect to the segment of agarose, causes agarose segment to be broken, agarose solution is thinning, in turn
The mechanical performance of ball after influence balling-up size and balling-up.Under conditions of mechanical stirring, water phase is dispersed in oily phase, is being collided
In merge again；- OH on agarose reacts under alkaline environment with 3- chlorine-2-hydroxyl propanesulfonate；While on agarose-
OH is crosslinked with the epoxychloropropane in organic phase to react (reaction equation is as follows), and slightly cross-linked water phase agarose may
It mixes again with next aqueous phase droplets, crosslinking continues；The additional amount of sodium hydroxide solution is too small, it is difficult to meet the chloro- 2- of 3-
Hydroxypropionate sodium and hydroxyl react and requirement of the epoxychloropropane cross-linking reaction to alkali.
Control 2 DEG C/min of cooling velocity is cooled to 10 DEG C or less to temperature in step d).
Ethanol solution described in step e) is 2.5-4.5 times of cured product volume；The NaOH solution is solidification
0.9-1.2 times of bulk product；The NaOH solution is 3.5-4.5mol/L.
During the cross-linking reaction of epoxychloropropane and agarose, epoxychloropropane be it is excessive, exist and do not hand over completely
The case where connection；Hydroxyl on one end and agarose of epoxychloropropane reacts (such as chemical equation 1, P represents agarose)
The other end does not react, and does not achieve the purpose that crosslinking.The cured product prepared is added in ethyl alcohol and sodium hydroxide solution,
In alkaline environment, one end that epoxychloropropane is not crosslinked reacts that (such as chemical equation 2, P represents fine jade with the hydroxyl of agarose
Lipolysaccharide)；
Volumes of aqueous ethanol is 2.5-4.5 times of cured product volume, and amount of alcohol is very little, cannot effectively disperse agarose
Promote the dispersion of alkali, if too many, the amount needs of alkali are correspondinglyd increase, and waste solvent；The NaOH solution of 3.5-4.5mol/L is added,
In an amount of from 0.9-1.2 times of cured product volume, NaOH solution concentration is too high, has destructiveness, NaOH to cured product skeleton
Solution concentration is too low, cannot effectively facilitate the cross-linking reaction of epoxychloropropane；The excessive concentration of NaOH solution volume is too low, unfavorable
In the cross-linking reaction of epoxychloropropane；The too small concentration of volume is too big, it is evenly dispersed in ball to be unfavorable for aqueous slkali, and then not
Conducive to the uniformity of crosslinking.
Key technology of the invention has at 3 points: key technology first is that the addition of 3- chlorine-2-hydroxyl propanesulfonate solid is suitable
Sequence.3- chlorine-2-hydroxyl propanesulfonate solid is added in the aqueous solution of agarose, is sufficiently mixed with the aqueous solution of agarose, is protected
Strong acid group has been demonstrate,proved in the uniformity of agar sugar ball inner part cloth, traditional strong acid ion group has been solved and is distributed in chromatography media
Inhomogenous problem.Key technology second is that strong acid functionalization, cross-linking reaction and balling-up carry out simultaneously, simplify reaction step, drop
Low cost.In the prior art, strong acid functionalization, cross-linking reaction and balling-up cannot carry out simultaneously.Because of strong acid functionalization and friendship
Connection reaction all must just can be carried out in alkaline environment, and at 70-75 DEG C, agarose skeleton is easy to be destroyed by aqueous slkali, existing
The addition of technology alkali is all excessive, therefore is infeasible；On the one hand the formation of agarose gel microsphere is to pass through agarose
On the other hand hydrogen bond action on skeleton is the crosslinked action by epoxychloropropane, prior art processes (alkali additional amount, temperature
Degree, reaction time) control unreasonable, the unsuccessful formation for influencing microballoon of crosslinking, alkali, epoxychloropropane and 3- chlorine-2-hydroxyl third
Excessive, the big influence hydrogen bond action of the ionic strength of formation is added in sodium sulfonate, and then influences the formation of microballoon.The present invention has passed through instead
It should match, the strict control in reaction time and temperature, be effectively controlled alkali to the destruction of agarose skeleton and ionic strength
The influence of hydrogen bond, thus the present invention is feasible.Key technology third is that the post-processing of reaction, due to added in cross-linking reaction
The epoxychloropropane entered be it is excessive, there are the hydroxyls above one end of epoxychloropropane and agarose to react in reaction,
And the responseless situation of the other end, responseless one end continues to have reacted with the hydroxyl on agarose in last handling process
At cross-linking process, the mechanical strength of agarose base chromatography media ensure that.
In conclusion the present invention uses the formation of 3- chlorine-2-hydroxyl propanesulfonate and epoxychloropropane to agarose microbeads
Process carries out functionalization and cross-linking reaction, and strong acid functionalization, cross-linking reaction and one step of balling-up carry out, and by testing crucial shadow
The control of the factor of sound prepares functionalization degree height, strong acid functional groups is evenly distributed, the microballoon of good mechanical property, simplifies its preparation
Technique saves the preparation cost.
Detailed description of the invention
Fig. 1 is the functional group structure of strong acid agarose chromatography medium；
Fig. 2 is the photo (amplification factor 100) that strong acid agarose interlayer analyses matter under optical microscopy；What the present invention obtained
Agarose gel microsphere surface is smooth, and ball-type is mellow and full, even particle size distribution；
Fig. 3 is cross-linked agarose gel microballoon and GE SP-Sepharose the FF stream of embodiment 1 and prior art preparation
The relation curve of speed and pressure；Reflect the pressure resistance for the agarose gel microsphere ratio GE SP-Sepharose FF that the present invention obtains
Performance is slightly good, and compared with the agarose gel microsphere of prior art preparation, mechanical strength is good.
Specific embodiment
Embodiment 1
A) prepared by water phase
The agarose solution (containing 10%KCl) that mass concentration is 4% is prepared, is heated to 92 DEG C, agarose is all molten
Solution is added 3- chlorine-2-hydroxyl propanesulfonate solid and stirs to dissolution, obtains water phase.The 3- chlorine-2-hydroxyl propanesulfonate and fine jade
The mass ratio of lipolysaccharide is 2:1.
B) prepared by suspension
Water phase is poured into 70 DEG C of Toluene oil phase, and oil is mutually 8:1 with the volume ratio of water phase, obtains suspension；Wherein in toluene
The polyvinyl acetate and oleic acid contained, quality are respectively the 2% and 0.8% of toluene quality；
C) strong acid functionalization and cross-linking reaction
Epoxychloropropane is added into step b) suspension, and the NaOH solution that mass concentration is 50% is added dropwise, and in 1h
Inside it is added dropwise；70 DEG C are maintained the temperature at, 3h is reacted, obtains cross-linking reaction product；The matter of the epoxychloropropane and agarose
Amount is than being 3:1；The volume of the 50%NaOH solution of dropwise addition is 2 times of volume of epoxychloropropane；
D) solidify
It by step c) cross-linking reaction product, is condensed, control 2 DEG C/min of cooling velocity to temperature is cooled to 8 DEG C；Repeatedly
The cured product that partial size is 100-300 mesh is prepared after cleaning.
E) it post-processes
The cured product prepared in step d) is weighed, is added in the ethanol solution of cured product volume three times, is added solid
Change the NaOH solution of the isometric 4mol/L of product, constant temperature stirs 12h at 50 DEG C, obtains product, i.e. strong acid agarose gel microsphere.
Embodiment 2
A) prepared by water phase
The agarose solution (containing 10%KCl) that mass concentration is 4% is prepared, is heated to 95 DEG C, agarose is all molten
Solution is added 3- chlorine-2-hydroxyl propanesulfonate solid and stirs to dissolution, obtains water phase.The 3- chlorine-2-hydroxyl propanesulfonate and fine jade
The mass ratio of lipolysaccharide is 2.5:1.
B) prepared by suspension
Water phase is poured into 70 DEG C of Toluene oil phase, oil is mutually 7:1 with the volume ratio of water phase, obtains suspension；Wherein toluene
In the polyvinyl acetate and oleic acid that contain, quality is respectively the 1.5% and 1% of toluene quality；
C) strong acid functionalization and cross-linking reaction
Epoxychloropropane is added into step b) suspension, and the NaOH solution that mass concentration is 50% is added dropwise, and in 1h
Inside it is added dropwise；70 DEG C are maintained the temperature at, 4h is reacted, obtains cross-linking reaction product；The matter of the epoxychloropropane and agarose
Amount is than being 3:1；The volume of the 50%NaOH solution of dropwise addition is 2.5 times of volume of epoxychloropropane；
D) solidify
It by step c) cross-linking reaction product, is condensed, control 2 DEG C/min of cooling velocity to temperature is cooled to 5 DEG C；Repeatedly
The cured product that partial size is 100-300 mesh is prepared after cleaning.
E) it post-processes
The cured product prepared in step d) is weighed, is added in the ethanol solution of cured product volume three times, is added solid
Change the NaOH solution of the isometric 4mol/L of product, constant temperature stirs 8h at 60 DEG C, obtains product, i.e. strong acid agarose gel microsphere.
Embodiment 3
A) prepared by water phase
The agarose solution (containing 10%KCl) that mass concentration is 4% is prepared, is heated to 93 DEG C, agarose is all molten
Solution is added 3- chlorine-2-hydroxyl propanesulfonate solid and stirs to dissolution, obtains water phase；The 3- chlorine-2-hydroxyl propanesulfonate and fine jade
The mass ratio of lipolysaccharide is 1.5:1.
B) prepared by suspension
Water phase is poured into 70 DEG C of Toluene oil phase, oil is mutually 6-8:1 with the volume ratio of water phase, obtains suspension；Wherein first
The polyvinyl acetate and oleic acid contained in benzene, quality are respectively the 1.8% and 1.2% of toluene quality；
C) strong acid functionalization and cross-linking reaction
Epoxychloropropane is added into step b) suspension, and the NaOH solution that mass concentration is 50% is added dropwise, and in 1h
Inside it is added dropwise；70 DEG C are maintained the temperature at, 6h is reacted, obtains cross-linking reaction product；The matter of the epoxychloropropane and agarose
Amount is than being 3:1；The volume of the 50%NaOH solution of dropwise addition is 2.5 times of volume of epoxychloropropane；
D) solidify
It by step c) cross-linking reaction product, is condensed, control 2 DEG C/min of cooling velocity to temperature is cooled to 2 DEG C；Repeatedly
The cured product that partial size is 100-300 mesh is prepared after cleaning；
E) it post-processes
The cured product prepared in step d) is weighed, is added in the ethanol solution of cured product volume three times, is added solid
Change the NaOH solution of the isometric 4mol/L of product, constant temperature stirs 16h at 40 DEG C, obtains product, i.e. strong acid agarose gel microsphere.
Embodiment 4
A) prepared by water phase
The agarose solution (containing 10%KCl) that mass concentration is 8% is prepared, is heated to 95 DEG C, agarose is all molten
Solution is added 3- chlorine-2-hydroxyl propanesulfonate solid and stirs to dissolution, obtains water phase；The 3- chlorine-2-hydroxyl propanesulfonate and fine jade
The mass ratio of lipolysaccharide is 1.8:1.
B) prepared by suspension
Water phase is poured into 80 DEG C of Toluene oil phase, oil is mutually 6.5:1 with the volume ratio of water phase, obtains suspension；Wherein first
The polyvinyl acetate and oleic acid contained in benzene, quality are respectively the 1% and 1% of toluene quality；
C) strong acid functionalization and cross-linking reaction
Epoxychloropropane is added into step b) suspension, and the NaOH solution that mass concentration is 50% is added dropwise, and in 1h
Inside it is added dropwise；75 DEG C are maintained the temperature at, 5h is reacted, obtains cross-linking reaction product；The matter of the epoxychloropropane and agarose
Amount is than being 3:1；The volume of the 50%NaOH solution of dropwise addition is 1.5 times of volume of epoxychloropropane；
D) solidify
It by step c) cross-linking reaction product, is condensed, control 2 DEG C/min of cooling velocity to temperature is cooled to 2 DEG C；Repeatedly
The cured product that partial size is 100-300 mesh is prepared after cleaning；
E) it post-processes
The cured product prepared in step d) is weighed, is added in 2.5 times of cured product volume of ethanol solution, is added solid
Change the NaOH solution of the 3.5mol/L of 0.9 times of volume of product, constant temperature stirs 10h at 45 DEG C, obtains product, i.e. strong acid Ago-Gel
Microballoon.
Embodiment 5
A) prepared by water phase
The agarose solution (containing 10%KCl) that mass concentration is 6% is prepared, is heated to 95 DEG C, agarose is all molten
Solution is added 3- chlorine-2-hydroxyl propanesulfonate solid and stirs to dissolution, obtains water phase；The 3- chlorine-2-hydroxyl propanesulfonate and fine jade
The mass ratio of lipolysaccharide is 2.2:1.
B) prepared by suspension
Water phase is poured into 75 DEG C of Toluene oil phase, oil is mutually 7.5:1 with the volume ratio of water phase, obtains suspension；Wherein first
The polyvinyl acetate and oleic acid contained in benzene, quality are respectively the 1.2% and 1.2% of toluene quality；
C) strong acid functionalization and cross-linking reaction
Epoxychloropropane is added into step b) suspension, and the NaOH solution that mass concentration is 50% is added dropwise, and in 1h
Inside it is added dropwise；It maintains the temperature at 70 DEG C, reacts 6h, obtain cross-linking reaction product；The epoxychloropropane and agarose
Mass ratio is 3:1；The volume of the 50%NaOH solution of dropwise addition is 3 times of volume of epoxychloropropane；
D) solidify
It by step c) cross-linking reaction product, is condensed, control 2 DEG C/min of cooling velocity to temperature is cooled to 4 DEG C；Repeatedly
The cured product that partial size is 100-300 mesh is prepared after cleaning；
E) it post-processes
The cured product prepared in step d) is weighed, is added in 4.5 times of cured product volume of ethanol solution, is added solid
Change the NaOH solution of the 4.5mol/L of 1.2 times of volumes of product, constant temperature stirs 16h at 40 DEG C, obtains product, i.e. strong acid Ago-Gel
Microballoon.
With reference to pertinent literature, the product of the product obtained using the present invention and prior art acquisition respectively adsorbs lysozyme
Performance is studied, and the results are shown in Table 1.The adsorbance of agarose gel microsphere prepared by the present invention is slightly above GE SP-
The adsorbance of Sepharose FF, hence it is evident that higher than the adsorbance of prior art preparation ball.
Table 1
Bibliography are as follows:
[1] Gao, D., Lin, D.Q., Yao, S.J.Measurement and correlation of protein
adsorption with mixed-mode adsorbents taking into account the influences of
salt concentration and pH[J].Journal of Chemical and Engineering Data,2006,51
(4):1205.
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Claims (7)
1. a kind of synthetic method of high adsorption capacity strong acid agar base chromatography media, which is characterized in that using agarose solution as water phase,
Toluene is oily phase, carries out function to the forming process of agarose microbeads using 3- chlorine-2-hydroxyl propanesulfonate and epoxychloropropane
Change and cross-linking reaction, are finally post-processed, obtain product.
2. a kind of synthetic method of high adsorption capacity strong acid agar base chromatography media according to claim 1, which is characterized in that
The specific steps are that:
A) prepared by water phase
The agarose solution that mass concentration is 4-8% is prepared, is heated to 90 DEG C or more, agarose all dissolves, and it is chloro- that 3- is added
2- hydroxypropionate sodium solid is stirred to dissolution, obtains water phase；
B) prepared by suspension
Water phase is poured into 70-80 DEG C of Toluene oil phase, oil is mutually 6-8:1 with the volume ratio of water phase, obtains suspension；
C) strong acid functionalization and cross-linking reaction
Epoxychloropropane is added into step b) suspension, and mass concentration is added dropwise and is 50% NaOH solution, and is dripped in 1h
It adds complete；70-75 DEG C is maintained the temperature at, 3-6h is reacted, obtains cross-linking reaction product；
D) solidify
By step c) cross-linking reaction product, is condensed, be cooled to 10 DEG C or less；It is 100- that partial size is prepared after being cleaned multiple times
The cured product of 300 mesh；
E) it post-processes
The cured product prepared in step d) is weighed, is added in ethanol solution, NaOH solution, perseverance at 40-60 DEG C is then added
Temperature stirring 8-16h, obtains product, i.e. strong acid agarose gel microsphere.
3. a kind of synthetic method of high adsorption capacity strong acid agar base chromatography media according to claim 1, which is characterized in that
The mass ratio of 3- chlorine-2-hydroxyl propanesulfonate and agarose described in step a) is 1.5-2.5:1；The agarose is water-soluble
Potassium chloride is added in liquid, and the mass concentration of potassium chloride is 10%.
4. a kind of synthetic method of high adsorption capacity strong acid agar base chromatography media according to claim 1, which is characterized in that
The polyvinyl acetate and oleic acid contained in Toluene oil phase in step b), quality are respectively the 1-2% and 0.8- of toluene quality
1.2%.
5. a kind of synthetic method of high adsorption capacity strong acid agar base chromatography media according to claim 1, which is characterized in that
The mass ratio of epoxychloropropane described in step c) and agarose is 3:1；The volume of the 50%NaOH solution of dropwise addition is epoxy
1.5-3 times of the volume of chloropropane.
6. a kind of synthetic method of high adsorption capacity strong acid agar base chromatography media according to claim 1, which is characterized in that
Control 2 DEG C/min of cooling velocity is cooled to 10 DEG C or less to temperature in step d).
7. a kind of synthetic method of high adsorption capacity strong acid agar base chromatography media according to claim 1, which is characterized in that
Ethanol solution described in step e) is 2.5-4.5 times of cured product volume；The NaOH solution is cured product volume
0.9-1.2 times；The NaOH solution is 3.5-4.5mol/L.
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