CN1059356C - Modified polysulphone super-filter membrane and its preparation and application - Google Patents
Modified polysulphone super-filter membrane and its preparation and application Download PDFInfo
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- CN1059356C CN1059356C CN94112724A CN94112724A CN1059356C CN 1059356 C CN1059356 C CN 1059356C CN 94112724 A CN94112724 A CN 94112724A CN 94112724 A CN94112724 A CN 94112724A CN 1059356 C CN1059356 C CN 1059356C
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Abstract
The present invention relates to a modified polysulfone ultrafiltration film which is prepared by carrying out a modification reaction to polysulfone with an acidylation-amination or chloromethylation-amination method. The ultrafiltration film has the following physical characteristics: the average pore diameter is from 20 to 80 mum; the pore density is 5*10<9> to 8*10<9> pore//cm<2>; the thickness of the skin layer of the film is from 5 to 12 mum; the thickness of a supporting layer is from 50 to 120 mum. The ultrafiltration film can be crosslinked with parenzyme under the crosslinked action of glutaraldehyde, and can also be treated through fixed carrying with parenzyme and prepared into a compatible ultrafiltration film through a diazo reactionat 0 to 4 DEG C. The film can be used for purified separation of trypsin inhibitors, and makes the purity of the inhibitors reach 98.5%.
Description
The present invention relates to a kind of membrane separation technique, specifically provide a kind of polysulphone super-filter membrane and preparation method thereof of modification.Simultaneously provide the method for utilizing this film to make the affinity ultrafiltration film again, this film can carry out purifies and separates to trypsin inhibitor.
Affine separation is owing to have very high specificity, good selectivity, high purifying multiple, so this technical development is very rapid, make multiple affine separation chromatography filler, be widely used in the research of the purifies and separates and the life science of bioengineering target product.Recently, along with affinity ligand is covalently bound on miillpore filter or the milipore filter have been appearred in film separation science develop rapidly, make affinity membrane, be applied on the purifies and separates of some large biological molecules such as enzyme, protein etc., used membrane material major part is cellulose membrane or cellulose blending film.Also have affinity ligand is covalently bound to (as polyacrylamide, poly-trimethylolpropene acid esters etc.) on some high polymer particles, adopt general milipore filter, large biological molecule is carried out purifies and separates, obtain effect preferably.Close with technology of the present invention, and people such as ALNguyen (BiotechnolBioeng, 1989,34:1186) adopt acryloyl chloride and aminobenzene carbonamidine and acrylamide under the condition of secluding air, to carry out liquid-phase bulk polymerization, make a kind of water miscible high polymer.This copolymer and trypsase are directly produced affinity interaction, made the water-soluble polymer of band trypsase affinity ligand.Utilize again that to hold back be that 100,000 daltonian general milipore filters carry out ultra-filtration and separation,, reach the purpose of purifies and separates trypsase or its inhibitor to remove other foreign protein.But adopt this method, toxicity such as used acryloyl chloride, benzenecarboximidamide are bigger when not only synthetic, and the formality complexity.Simultaneously, because copolymer and trypsase are direct-connected, do not have spacerarm, so separative efficiency being low again, only is 90% to the highest tryptic rejection, and binding capacity is 10.5mg/ml.
The purpose of this invention is to provide a kind of modified polysulphone super-filter membrane and preparation method.The operation of this technology of preparing is simple and easy, and production cost is low, and the product yield height is easy on the milipore filter of gained that enzyme or protein are carried out the immobilized affinity ultrafiltration film of making and can be used for the technical pure trypsin inhibitor is carried out purifies and separates.
Polysulphone super-filter membrane provided by the invention is that the employing polysulfones is that raw material obtains a kind of milipore filter through the modification processing, it is characterized in that being connected in this filter membrane band active function groups-NH
2Spacerarm possible constructions formula I or II represent:
The physical property of above-mentioned modified polysulphone super-filter membrane is: average pore size is at 20～80 μ m, and hole density is 5 * 10
9～8 * 10
9Hole/cm
2, the skin thickness of film is 5～12 μ m, supporting layer thickness is 50～120 μ m.
The preparation method of modified polysulphone super-filter membrane of the present invention mainly contains two steps: 1. the chemical modification of pair polysulfones raw material is handled and the film forming of 2. milipore filters, it is characterized in that the chemical modification of polysulfones can select one of following two kinds of synthetic methods to carry out:
1) acidylate-amination method:
Acidylate-amination method is to be raw material with the polysulfones; under the catalytic action of alchlor; under 60～80 ℃; make itself and paranitrobenzoyl chloride carry out acylation reaction (1); the gained product is 40～60 ℃ times and hydrazine (diamine) carries out aminating reaction (2), makes and is connected to band polar functional group-NH on the polysulfones molecule
2The spacer groups of base, its reaction equation is:
In the above-mentioned reaction (1), the addition of catalyst be polysulfones heavy 1～8%, the addition of reactant paranitrobenzoyl chloride (molal quantity) should be 1～4 times of polysulfones, reaction time is 1～6 hour, in the reaction (2), the addition of reactant hydrazine should be 1～5 times of polysulfones, and the reaction time is 2～6 hours.After reaction (2) finished, the conversion ratio of polysulfones was 30～50%.
2) chloromethylation-amination method
Chloromethylation-amination method is that polysulfones is carried out chloromethylation (3) with chloromethyl ether earlier down at 20～30 ℃, again product and p-phenylenediamine (PPD) is carried out aminating reaction (4) under 55～65 ℃, makes the chemical modification polysulfones of band anilino-, and its reaction equation is
In the above-mentioned reaction (3), the addition of reactant chloromethyl ether (molal quantity) should be 1～4 times of polysulfones, reaction time is 1～6 hour, the addition of the middle reactant p-phenylenediamine (PPD) of reaction (4) should be 1～4 times of polysulfones, reaction time is 2～6 hours, and reaction (4) finishes the conversion ratio of back polysulfones 40～75%.
The film forming of milipore filter of the present invention, available phase inversion, technology is carried out routinely, it is characterized in that also can carrying out film preparation by following step:
With the chemical modification polysulfones under 45～60 ℃; by continuously evenly stirring and dissolving at N; in the dinethylformamide; fully after the uniform dissolution; under the protection of nitrogen vapour lysate is filtered, scraper striking film forming is used in the degassing then on glass plate; after in air, exposing 20～50 seconds, immerse film forming in the redistilled water that contains 2～10%NaCl.Make average pore size between 20～80 μ m through this process, hole density is 5 * 10
9～8 * 10
9Hole/cm
2, the skin thickness of film is 5～12 μ m, the back strut layer thickness is the milipore filter of 50～120 μ m.
Have active group-NH as above-mentioned by the prepared modified polysulphone super-filter membrane that goes out of method provided by the present invention
2, since can be under the effect of glutaraldehyde crosslinked with large biological molecule such as trypsase, also can by diazo-reaction under 0～4 ℃,, make affine separating medium (affinity ultrafiltration film) with its generation covalent bond.This affinity ultrafiltration film can be used for large biological molecule, carries out purifies and separates as tryptose ester inhibitor, below by example technology of the present invention is given to illustrate further.
Example 1, the preparation 1 of modified polysulphone super-filter membrane
Adopting acidylate-amination method that polysulfones is carried out modification handles; its process is as follows: PS-DL (1) (η=0.57) polysulfones that produces with Dalian first plastic molding and processing plant is a raw material; alchlor is catalyst (addition be polysulfones weight 5%); under 70 ℃; make itself and paranitrobenzoyl chloride react (addition is 3 times of polysulfones mole gram number); react after 2 hours, isolate product.Be under 50 ℃,, reacted 4 hours, isolate reactant, washed product with product hydrazine reaction (the hydrazine addition is 4 times of polysulfones mole gram number).The conversion ratio of its polysulfones reaches 45%, utilizes conventional phase inversion to make modified polysulphone super-filter membrane with above-mentioned product, and its film thickness is 100 μ m, and average pore size is the modified polysulphone super-filter membrane 1 of the band finger-like pore structure of 40 μ m.
The preparation 2 of example 2 modified polysulphone super-filter membranes
Adopting chloromethylation-amination method that polysulfones is carried out modification handles, its process is as follows: PS-DL (1) (η=0.57) polysulfones that produces with Dalian first plastic molding and processing plant is a raw material, react (the chloromethyl ether addition is 4 times of polysulfones molal quantity) with chloromethyl ether earlier down at 25 ℃, reacted 1 hour, again with reactant and p-phenylenediamine (PPD) (addition is 3 times of polysulfones molal quantity) reaction, reacted 4 hours down at 60 ℃, separated product washs, and the conversion ratio of its polysulfones is 70%.
Utilize above-mentioned modification polysulfones to be raw material, under 50～60 ℃, be dissolved in N, in the N-dimethylformamide (weight ratio of polysulfones and dimethylformamide is 8～15%), at N
2Under the gas lysate is filtered, the degassing then, on glass plate with scraper striking film forming, in air, expose 40～50 seconds after, immerse film forming in the second distillation aqueous solution of 10%NaCl, its thickness is the modified polysulphone super-filter membrane of the band finger-like pore structure of 105 μ m.
Example 3, the preparation of the affine filter membrane of modification polysulfones
Use-case 1 and 2 prepared modified polysulphone super-filter membranes, utilize the diazotising method to carry out the immobilized reaction of tryptose ferment. its reaction temperature is 4 ℃, use the phosphate buffer (0.05mol/l of tryptic concentration as 0.5mg/ml, PH8.1), be reactant liquor, carry out immobilized reaction 4 hours, after reaction finishes, use deionized water successively, 0.1 the NaCl solution of～1.0mol/L, deionized water repeated multiple times cleaning filter membranes, up to cleaning fluid till trypsase can not detect in the 280mm place of ultraviolet specrophotometer.Immobilized enzyme membrane is measured: immobilized 13.5mg trypsase on every gram modification PS membrane on the affinity membrane of example 1, supported tryptic relative vigor is 13000u/g.Immobilized 15mg trypsase on every gram modification PS membrane on the affinity membrane of example 2, supported tryptic relative vigor is 18200u/g,
The application of the affine filter membrane of example 4 modification polysulfones
The affine filter membrane that utilizes example 3 to make, be assembled in the membrane separator, and use this separator, the technical pure soybean trypsin inhibitor is carried out purifying, purification condition is: the slow middle liquid of phosphoric acid that with concentration is 0.2mg/ml technical pure soybean trypsin inhibitor, flow with 0.5mol/min cycles through affinity membrane separator, fully washs the film storehouse with 0.05mol/L phosphate buffer and deionized water respectively after 1 hour, in flowing out liquid, can not detect the existence of trypsin inhibitor till.Urea liquid with 6mol/L passes through affinity membrane separator under same flow velocity again, collects eluent, analyzes with high performance liquid chromatography, and the purity of its trypsin inhibitor can reach 98.5%, has improved 68 times than former industrial trypsin inhibitor purity.
By above-mentioned example, the preparation method that modification polysulfones provided by the present invention plays filter membrane is simple, reaction condition is easy to control, and prepared milipore filter is easy to tryptic immobilized, and obtains the practical milipore filter that can be used for the purifies and separates trypsin inhibitor.
Claims (4)
1. a modified polysulphone super-filter membrane is characterized in that being connected in this filter membrane band active function groups-NH
2Spacer groups, represent with structural formula I or II:
2. according to the described milipore filter of claim 1, it is characterized in that this filter membrane has following physical property: average pore size: 20～80 μ m; Hole density: 5 * 10
9～8 * 10
9Hole/cm
2, the skin thickness of film is 5～12 μ m; Supporting layer thickness: 50～120 μ m.
3. preparation method according to the described modified polysulphone super-filter membrane of claim 1 comprises modification and two key steps of film forming of polysulfones, and it is special gives birth to the modification that is polysulfones and adopt acidylate-amination method or chloromethylation-amination method to carry out; It is so-called that acidylate-the amination method is that polysulfones is under the catalytic action of alchlor; in 60～80 ℃; make itself and paranitrobenzoyl chloride carry out acylation reaction, the gained product is 40～60 ℃ times and hydrazine carries out aminating reaction, makes and is connected to band polar functional group-NH on the polysulfones molecule
2The spacer groups of base;
So-called chloromethylation-amination method is that polysulfones carries out chloromethylation with chloromethyl ether earlier down at 20～30 ℃, again reactant and p-phenylenediamine (PPD) is carried out aminating reaction under 55～65 ℃, makes the modification polysulfones of band anilino-.
4. purifies and separates that is used for trypsin inhibitor according to the described modified polysulphone super-filter membrane of claim 1, its special giving birth to is to carry out crosslinked with trypsase this milipore filter under the effect of glutaraldehyde earlier, or by diazo-reaction 0～4 ℃ down and trypsase immobilized, make affine filter membrane; Be assembled in this affine filter membrane and be used on the membrane separator technical pure trypsin inhibitor is carried out purifies and separates.
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|---|---|---|---|
| CN94112724A CN1059356C (en) | 1994-12-28 | 1994-12-28 | Modified polysulphone super-filter membrane and its preparation and application |
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|---|---|---|---|
| CN94112724A CN1059356C (en) | 1994-12-28 | 1994-12-28 | Modified polysulphone super-filter membrane and its preparation and application |
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|---|---|
| CN1125635A CN1125635A (en) | 1996-07-03 |
| CN1059356C true CN1059356C (en) | 2000-12-13 |
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| CN1978038B (en) * | 2005-12-09 | 2010-05-05 | 中国科学技术大学 | Method for preparing homogeneous-phase anion exchange film |
| CN100395008C (en) * | 2006-03-31 | 2008-06-18 | 华南理工大学 | A hydrophilic-hydrophobic bipolar composite membrane and method for preparing same |
| KR100813249B1 (en) | 2006-10-31 | 2008-03-13 | 삼성에스디아이 주식회사 | Polysulfone, electrolyte membrane using the same, and fuel cell using the same |
| CN101100484B (en) * | 2007-06-12 | 2011-02-09 | 东华大学 | Method for fast purifying cysteine protease inhibitor |
| CN102935336A (en) * | 2012-11-26 | 2013-02-20 | 天津工业大学 | Manufacturing method of blended flat affinity filter membrane |
| CN108939957B (en) * | 2018-08-01 | 2020-11-27 | 中南大学湘雅医院 | Preparation method of Avacopan modified dialysis membrane |
| CN110943237B (en) * | 2018-09-21 | 2021-09-28 | 中国科学院大连化学物理研究所 | Application of ion-conducting membrane in flow battery |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63178806A (en) * | 1987-10-29 | 1988-07-22 | Toray Ind Inc | Production of semipermeable compound film having high performance |
| JPH05220389A (en) * | 1992-02-12 | 1993-08-31 | Showa Denko Kk | Affinity separation membrane |
| EP0569229A1 (en) * | 1992-05-05 | 1993-11-10 | W.R. Grace & Co. | High efficiency removal of low density lipoprotein-cholesterol from whole blood |
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1994
- 1994-12-28 CN CN94112724A patent/CN1059356C/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63178806A (en) * | 1987-10-29 | 1988-07-22 | Toray Ind Inc | Production of semipermeable compound film having high performance |
| JPH05220389A (en) * | 1992-02-12 | 1993-08-31 | Showa Denko Kk | Affinity separation membrane |
| EP0569229A1 (en) * | 1992-05-05 | 1993-11-10 | W.R. Grace & Co. | High efficiency removal of low density lipoprotein-cholesterol from whole blood |
Non-Patent Citations (1)
| Title |
|---|
| BIOTECHNOL BIOENG 34 1989.1.1 A.L.NGUYEN * |
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| CN1125635A (en) | 1996-07-03 |
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