CN105753976A - Method for purifying ulinastatin based on cation exchange resin - Google Patents
Method for purifying ulinastatin based on cation exchange resin Download PDFInfo
- Publication number
- CN105753976A CN105753976A CN201610323431.0A CN201610323431A CN105753976A CN 105753976 A CN105753976 A CN 105753976A CN 201610323431 A CN201610323431 A CN 201610323431A CN 105753976 A CN105753976 A CN 105753976A
- Authority
- CN
- China
- Prior art keywords
- ulinastatin
- cation exchange
- exchange resin
- membrane
- filtrate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/81—Protease inhibitors
- C07K14/8107—Endopeptidase (E.C. 3.4.21-99) inhibitors
- C07K14/811—Serine protease (E.C. 3.4.21) inhibitors
- C07K14/8114—Kunitz type inhibitors
Abstract
The invention belongs to the technical field of medicines, and in particular relates to a method for purifying ulinastatin based on cation exchange resin. The method comprises the following steps: mixing chitosan and alginate with primarily filtered urine, performing ultrafiltration, and purifying the trapped fluid obtained through ultrafiltration through cation exchange resin and ultrafiltration in sequence. By adoption of the method for purifying ulinastatin based on the cation exchange resin, the purification process is shortened, the yield and the purity of the ulinastatin are high, the yield of the ulinastatin is 75% or above, and the specific activity of the ulinastatin is up to 4750 IU/mg protein.
Description
Technical field
The invention belongs to pharmaceutical technology field, be specifically related to a kind of method based on cation exchange resin purification ulinastatin.
Background technology
Ulinastatin system separates a kind of acidoglycoprotein being purified into from male urine.Very wide zymogram is pressed down, so trypsin, phospholipase A can be suppressed simultaneously owing to the Liang Ge active function district in structure is respectively provided with2, hyaluronidase, the multiple hydrolytic enzyme such as elastoser activity；And, the low molecular weight compositions that ulinastatin is decomposed to form also has the very strong effect suppressing hydrolytic enzyme.Ulinastatin is not only the protease inhibitor of a kind of wide spectrum, also have suppress myocardial depressant factor generation, recurrent state when improving shock, stablize the function such as release of lysosome membrane, suppression inflammatory mediator.A large amount of clinical test results show, ulinastatin has important effect in treating acute pancreatitis, auxiliary treatment shock, improving operation to stimulate the immunologic function degression caused, especially minimizing complication of extracorporeal circulation etc., plus itself deriving from human body, non-immunogenicity, safety is high.
But owing in urine, the content of ulinastatin is less, it separates and purifying process becomes the key factor restricting its application.
About separation and the purification process of ulinastatin, domestic report is less.Through retrieval, it has been disclosed that only 21 sections of Chinese patent application document.The method that Chinese patent application CN2015108204251 " a kind of resin regeneration improves the method for post effect purification ulinastatin and improves the pharmaceutical composition of ulinastatin solubility " and CN2014108077092 " a kind of resin regeneration improves the method for post effect ulinastatin and the pharmaceutical composition containing ulinastatin " disclosed in the Kang Yuan pharmaceutcal corporation, Ltd of Qingdao all utilizes the method that anion-exchange column, affinity column and ultrafiltration combine that ulinastatin crude product is purified.The applicant adopts anion-exchange column, ultrafiltration and metal chelating column, drainage column and gel column to combine in patent documentation CN2006100002002 " ulinastatin of purification and preparation method thereof and the pharmaceutical composition containing this ulinastatin " before and ulinastatin crude product is purified.
The purification process of ulinastatin disclosed above all adopts repeatedly resin column to be purified.But for the urine of certain volume, purifying process is more complicated, can improve the purity of ulinastatin to a certain extent, and the yield of ulinastatin is affected.How to coordinate purifying process and the relation of its yield and purity of ulinastatin, become scientific research personnel's problem demanding prompt solution.
Cation exchange resin, as a kind of conventional purified material, can be used for the enrichment of the purification of material, separation and product.Difference according to purposes, the resin of optional different types of structure.Such as, " strong cation-exchanging resin application in protein separation " (" the Ningxia engineering " that Bu Chunmiao et al. delivered in 2006, in March, 2006, the 5th volume the 1st phase) namely it is prepared for the good strong cation-exchanging resin of a kind of hydrophilic and for Purification of Lysozyme from Ovum Gallus domesticus album and from Cor Sus domestica, extracts cytochrome G5 and achieve good effect.
But up to now, there are no the relevant report of the purification that cation exchange resin is used for ulinastatin.
Summary of the invention
It is an object of the invention to for the above-mentioned problems in the prior art, a kind of method based on cation exchange resin purification ulinastatin is provided,, while the purity ensureing ulinastatin and yield, to shorten its purifying process, reach to save the purpose of cost, minimizing time and human cost.
In order to realize foregoing invention purpose, the invention provides a kind of method based on cation exchange resin purification ulinastatin, it specifically comprises the following steps that
1) take clarification urine, regulate pH to 6-6.5, add silica gel, stirring, filter, obtain filtrate, after regulating filtrate pH to 3-5, add chitosan and alginate, stirring, obtain mixed liquor 1, be 5 × 10 by mixed liquor 1 with molecular cut off4-7×104Ultrafilter membrane 1 ultrafiltration obtain trapped fluid 1；
2) regulate the pH to 7-8 of trapped fluid 1, filter, obtain filtrate 1, upper cation exchange resin column after filtrate 1 is adjusted pH to 4-8, then with eluent 1 eluting, collect effluent and obtain effluent A；
3) it is 3 × 10 by effluent A molecular cut off4Ultrafilter membrane 2 ultrafiltration obtain trapped fluid 2；
4) by trapped fluid 2 lyophilization, pure ulinastatin is obtained.
Preferably, the method based on cation exchange resin purification ulinastatin provided by the invention, it specifically comprises the following steps that
1) take clarification urine, regulate pH to 6-6.5, with silica gel: the ratio of urine=8-12kg/t adds silica gel, stirring, filters, obtains filtrate, regulate filtrate pH to 3-5, with chitosan: the ratio of urine=8-12kg/t adds chitosan in filtrate, with weight ratio for alginate: the ratio of chitosan=20-50% adds alginate, stirring, obtain mixed liquor 1, mixed liquor 1 is 0.1-0.7MPa at pressure, when temperature is 4-30 DEG C, is 5 × 10 with molecular cut off4-7×104Ultrafilter membrane 1 ultrafiltration obtain trapped fluid 1；
2) pH to 7-8 of trapped fluid 1 is regulated, filter, obtain filtrate 1, after filtrate 1 is adjusted pH to 4-8, the speed with 80-120mL/min flows through cation exchange resin, collect effluent, above-mentioned effluent is flow through cation exchange resin with the speed of 80-120mL/min again, repeats 2-3 time, collect effluent and obtain effluent A；
3) by effluent A when pressure is 0.1-0.5MPa, pH=7-8, it is 3 × 10 with molecular cut off4Ultrafilter membrane 2 ultrafiltration obtain trapped fluid 2；
4) by trapped fluid 2 lyophilization, pure ulinastatin is obtained.
Wherein, the application step 2) in used cation exchange resin before the use routinely step carry out pretreatment, such as, sequentially include the following steps: and use distilled water immersion 24h, and it is washed till the clarification of water liquid with distilled water, incline and after anhydrating, add 1mol/LNaOH solution soaking 24h, be washed to neutrality, add 1mol/LHCl solution soaking 24h, wash with water to neutrality.
When two kinds of mixed with resin use, respectively mixing after the means pretreatment routinely of two kinds of resins is used.
Cation exchange resin before use, balances resin column with eluent.
Chitosan, obtains through deacetylation for chitin, the difference according to deacetylation degree, is divided into water-soluble chitosan and water-insoluble chitosan, it is preferable that be the chitosan more than 90% for deacetylation, and molecular weight is 2-5 ten thousand.
Preferably, described ultrafilter membrane is the composite membrane of a kind of or at least two in poly (ether sulfone) film, polypropylene screen, regenerated cellulose film, cellulose acetate membrane, polysulfone membrane and polyvinylidene fluoride film；The more preferably composite membrane of poly (ether sulfone) film and cellulose acetate membrane.
Preferably, the structure of described ultrafilter membrane is the one in hollow fiber form, flat, tubular type and rolling；More preferably inner pressed hollow fiber ultrafiltration membrane.
Preferably, described cation exchange resin is a kind of mixing in AmberliteIR120 and Amberlite200, AmberliteIRA118, AmberliteIRC84, Amberjet1200 and AmberliteIRC54；
More preferably AmberliteIR120 and Amberlite200, AmberliteIRA118, one in AmberliteIRC84, Amberjet1200 and AmberliteIRC54 is with mass ratio for (1-5): 1 mixing.
Preferably, step 2) in balance described cation exchange resin column solution be phosphate buffer or Tris-HCl buffer.
Under faintly acid, neutrality or alkali condition (pH=4--8), acidic protein ulinastatin is electronegative, will not with cation exchange resin generation exchange interaction, and the little Molecular Adsorption of impurity is on resin column, thus separating with ulinastatin.
Compared with prior art, the provided herein method separating purification ulinastatin has the advantage that (1) enriches the way of purification of ulinastatin, shorten purifying process, the crude separation step of the ulinastatin of the application substantially increases organic efficiency and the purity of ulinastatin, shortens the technological process of whole purification step.
(2) improve the response rate and the activity of ulinastatin, the response rate of ulinastatin reaches more than 75%；Rate activity reaches 4750IU/mg albumen, and (1 IU is defined as: with BAEE for substrate, when 25 degree, per minute makes Δ A253The amount of the ulinastatin needed for increase by 0.001).
Detailed description of the invention
Below in conjunction with embodiment, the present invention is further elaborated.These embodiments are only for illustrative purposes, and do not limit the scope of the invention and essence.Wherein, resin used herein is commercial.
Embodiment 1
1) take 1t and clarify urine, regulate pH to 6, add the silica gel of 8kg, stirring, with kieselguhr for media filtration, filtrate, regulate filtrate pH to 3, (molecular weight of chitosan is 50,000 to add the chitosan of 8kg in filtrate, deacetylation is 93%) and 0.4kg potassium alginate (molecular weight is 50,000), stirring 1h, obtains mixed liquor 1, by mixed liquor 1 pressure to be 0.1MPa, temperature be 30 DEG C, molecular cut off be 7 × 104When ultrafiltration, obtain trapped fluid 1；
2) pH to 7.5 of trapped fluid 1 is regulated, kieselguhr is media filtration, obtain filtrate 1, flow through cation exchange resin (AmberliteIR-120Na:AmberliteIRC54 mass ratio mixes for 5: 1) with the speed of 80mL/min after filtrate 1 is adjusted pH to 4, collect effluent and obtain effluent A；
3) be 0.2MPa, pH=7.2, molecular cut off by effluent A at pressure it is 3 × 104When ultrafiltration, obtain trapped fluid 2；
4) by trapped fluid 2 lyophilization, pure ulinastatin is obtained.
Wherein, in the present embodiment, ultrafilter membrane used is poly (ether sulfone) film and cellulose acetate membrane active layer mode all upwards is set up in parallel the composite membrane obtained, membrane structure is inner pressed doughnut structure, and other concrete structure parameter of ultrafilter membrane and operating procedure are configured by the routine techniques means of those skilled in the art.
Buffer solution used by balance cation exchanger resin is sodium phosphate buffer system.
Regulating the solution used by solution ph is acetic acid and ammonia.
The ulinastatin that the present embodiment obtains is white powder, and adopting the HPLC assay method that pharmacopeia is recommended to record purity is 99.5%, and the yield of ulinastatin is 85%, and Rate activity is 4750IU/mg albumen.
Embodiment 2
1) take 1t and clarify urine, regulate pH to 6.5, add the silica gel of 12kg, stirring, with kieselguhr for media filtration, filtrate, regulate filtrate pH to 4.5, (molecular weight of chitosan is 30,000 to add the chitosan of 12kg in filtrate, deacetylation is 99%) and 0.24kg sodium alginate (molecular weight is 50,000), stirring 1.5h, obtains mixed liquor 1, by mixed liquor 1 pressure to be 0.7MPa, temperature be 4 DEG C, molecular cut off be 5 × 104When ultrafiltration, obtain trapped fluid 1；
2) pH to 8 of trapped fluid 1 is regulated, filter, obtain filtrate 1, cation exchange resin AmberliteIR120Na is flow through with the speed of 100mL/min after filtrate 1 is adjusted pH to 5, collect effluent, above-mentioned effluent is flow through above-mentioned cation exchange resin AmberliteIR120Na with the speed of 80mL/min again, collects effluent and obtain effluent A；
3) be 0.5MPa, pH=8, molecular cut off by effluent A at pressure it is 3 × 104When, ultrafiltration obtains trapped fluid 2；
4) by trapped fluid 2 lyophilization, pure ulinastatin is obtained.
Wherein, ultrafilter membrane used by the present embodiment is poly (ether sulfone) film, and membrane structure is tubular ultra-filtration membrane, and ultrafilter membrane concrete structure parameter and operating procedure are with embodiment 1.
Regulating the solution used by solution ph is hydrochloric acid and sodium hydroxide.
The ulinastatin that the present embodiment obtains is white powder, and adopting and recording purity with the method for embodiment 1 is 99.2%, and the yield of ulinastatin is 79%, and Rate activity is 3700IU/mg albumen.
Buffer solution used by balance cation exchanger resin is potassium phosphate buffer system.
Embodiment 3
1) take 1t and clarify urine, regulate pH to 6.5, add the silica gel of 12kg, stirring, with kieselguhr for media filtration, filtrate, regulate filtrate pH to 5, (molecular weight of chitosan is 30,000 to add the chitosan of 10kg in filtrate, deacetylation is 95%) and 0.4kg potassium alginate (molecular weight is 50,000), stirring 2, obtains mixed liquor 1, by mixed liquor 1 pressure to be 0.1MPa, temperature be 25 DEG C, molecular cut off be 6 × 104When ultrafiltration, obtain trapped fluid 1；
2) pH to 7 of trapped fluid 1 is regulated, filter, obtain filtrate 1, cation exchange resin (AmberliteIR-120Na:Amberlite200 mass ratio mixes) is flow through for 5: 1 with the speed of 120mL/min after filtrate 1 is adjusted pH to 6, collect effluent, above-mentioned effluent is flow through above-mentioned cation exchange resin with the speed of 120mL/min again, repeats 2 times, collect effluent and obtain effluent A；
3) be 0.2MPa, pH=7.2, molecular cut off by effluent A at pressure it is 3 × 104When, ultrafiltration obtains trapped fluid 2；
4) by trapped fluid 2 lyophilization, pure ulinastatin is obtained.
Wherein, in the present embodiment ultrafilter membrane used be poly (ether sulfone) film, polyvinylidene fluoride film and cellulose acetate membrane by poly (ether sulfone) film, polyvinylidene fluoride film active layer all upwards, be placed in the cellulose acetate membrane active layer in intermediate layer downward in the way of the composite membrane obtained is set, membrane structure is inner pressed doughnut structure, and ultrafilter membrane concrete structure parameter and operating procedure are with embodiment 1.
Regulating the solution used by solution ph is sulphuric acid and sodium bicarbonate.
The ulinastatin that the present embodiment obtains is white powder, and adopting and recording purity with the method for embodiment 1 is 99.7%, and the yield of ulinastatin is 75%, and Rate activity is 3900IU/mg albumen.
Buffer solution used by balance cation exchanger resin is Tris-HCl buffer system.
Embodiment 4
1) ulinastatin roughing weighs the 1tpH clarification urine less than 6.5, is stirred continuously, and is slowly added to 16.5kg chitin, and measures with accurate pH test paper, regulates urine pH to 6.0；With ammonia eluting 1h after absorption completely, then through 3.5kg ammonium sulfate precipitation, overnight precipitation, centrifugal it is placed on vacuum drying in the vacuum desiccator making water absorbing agent with phosphorus pentoxide, namely obtains ulinastatin crude product after drying；
2) weigh ulinastatin crude product 1kg, dissolve with the acetate buffer of the 0.3mol/L of 8.5L, after stirring pressure to be 0.7MPa, temperature be 25 DEG C, molecular cut off be 5 × 104When ultrafiltration, obtain trapped fluid 1；
3) pH to 8 of trapped fluid 1 is regulated, filter, obtain filtrate 1, styrene type cation exchange resin AmberliteIR120Na is flow through with the speed of 100mL/min after filtrate 1 is adjusted pH to 7, collect effluent, above-mentioned effluent is flow through above-mentioned cation exchange resin with the speed of 120mL/min again, repeats 3 times, collect effluent and obtain effluent A；
4) be 0.5MPa, pH=8, molecular cut off by effluent A at pressure it is 3 × 104When, ultrafiltration obtains trapped fluid 2；
5) by trapped fluid 2 lyophilization, pure ulinastatin is obtained.
Wherein, the present embodiment regulates the solution used by solution ph is nitric acid and triethylamine.
The ulinastatin that the present embodiment obtains is pale yellow powder, and adopting and recording purity with the method for embodiment 1 is 97.5%, and the yield of ulinastatin is 61%, and Rate activity is 4750IU/mg albumen.
Buffer solution used by balance cation exchanger resin is Tris-HCl buffer system.
Embodiment 5
1) take 1t and clarify urine, regulate pH to 6, add the silica gel of 8kg, stirring, with kieselguhr for media filtration, filtrate, regulate filtrate pH to 3, (molecular weight of chitosan is 50,000 to add the chitosan of 8kg in filtrate, deacetylation is 93%) and 0.4kg potassium alginate (molecular weight is 50,000), stirring 1h, obtains mixed liquor 1, by mixed liquor 1 pressure to be 0.1MPa, temperature be 25 DEG C, molecular cut off be 7 × 104When ultrafiltration, obtain trapped fluid 1；
2) pH to 7.5 of trapped fluid 1 is regulated, kieselguhr is media filtration, obtain filtrate 1, cation exchange resin (AmberliteIR-120Na:AmberliteIRC54 mass ratio mixes) is flow through for 5: 1 with the speed of 80mL/min after filtrate 1 is adjusted pH to 8, collect effluent, above-mentioned effluent is flow through above-mentioned cation exchange resin with the speed of 120mL/min again, repeats 2 times, collect effluent and obtain effluent A；
3) be 0.2MPa, pH=7.2, molecular cut off by effluent A at pressure it is 3 × 104When ultrafiltration, obtain trapped fluid 2；
4) in trapped fluid 2, the ethanol that volumetric concentration is 95% of addition 2 times precipitates, and obtains precipitate 1；
5) by precipitate 1 drying under reduced pressure, pure ulinastatin is obtained.
Wherein, the ulinastatin that the present embodiment obtains is white powder, and adopting and recording purity with the method for embodiment 1 is 99.5%, and the yield of ulinastatin is 73%, and Rate activity is 4200IU/mg albumen.
Buffer solution used by balance cation exchanger resin is Tris-HCl buffer system.
Embodiment 6
1) take 1t and clarify urine, regulate pH to 6, add the silica gel of 8kg, stirring, with kieselguhr for media filtration, obtain filtrate, regulate filtrate pH to 3, (molecular weight of chitosan is 50,000 to add the chitosan of 8kg in filtrate, deacetylation is 93%) and 0.4kg potassium alginate (molecular weight is 50,000), stir 1h, stand 1h, centrifugal 0.5h, obtains lower sediment thing 1；
2) in precipitate 1, add ammonium sulfate buffer solution, regulate pH to 6.5, pressure to be 0.1MPa, temperature be 30 DEG C, molecular cut off be 7 × 104When ultrafiltration, obtain trapped fluid 1；
3) pH to 7.5 of trapped fluid 1 is regulated, kieselguhr is media filtration, obtain filtrate 1, cation exchange resin (AmberliteIR-120Na:AmberliteIRC54 mass ratio mixes) is flow through for 5: 1 with the speed of 80mL/min after filtrate 1 is adjusted pH to 4.5, collect effluent, above-mentioned effluent is flow through above-mentioned cation exchange resin with the speed of 120mL/min again, repeats 2 times, collect effluent and obtain effluent A；
4) be 0.2MPa, pH=7.2, molecular cut off by effluent A at pressure it is 3 × 104When ultrafiltration, obtain trapped fluid 2；
5) by trapped fluid 2 lyophilization, pure ulinastatin is obtained.
Wherein, the ulinastatin that the present embodiment obtains is white powder, and adopting and recording purity with the method for embodiment 1 is 99.4%, and the yield of ulinastatin is 70%, and Rate activity is 3741IU/mg albumen.
Embodiment 7
1) take 1t and clarify urine, regulate pH to 6, add the silica gel of 8kg, stirring, with kieselguhr for media filtration, obtain filtrate, regulate filtrate pH to 3, (molecular weight of chitosan is 50,000 to add the chitosan of 8kg in filtrate, deacetylation is 93%) and 0.4kg potassium alginate (molecular weight is 50,000), stir 1h, stand 1h, centrifugal 0.5h, obtains lower sediment thing 1；
2) in precipitate 1, add ammonium sulfate buffer solution, regulate pH to 6.5, pressure to be 0.1MPa, temperature be 10 DEG C, molecular cut off be 7 × 104When ultrafiltration, obtain trapped fluid 1；
3) pH to 7.5 of trapped fluid 1 is regulated, kieselguhr is media filtration, obtain filtrate 1, cation exchange resin (AmberliteIR-120Na:AmberliteIRC54 mass ratio mixes) is flow through for 5: 1 with the speed of 80mL/min after filtrate 1 is adjusted pH to 5.5, collect effluent, above-mentioned effluent is flow through above-mentioned cation exchange resin with the speed of 80mL/min again, repeats 3 times, collect effluent and obtain effluent A；
4) be 0.2MPa, pH=7.2, molecular cut off by effluent A at pressure it is 3 × 104When ultrafiltration, obtain trapped fluid 2；
5) in trapped fluid 2, the ethanol that volumetric concentration is 95% of addition 2 times precipitates, and obtains precipitate 2；
6) by precipitate 2 drying under reduced pressure, pure ulinastatin is obtained.
Wherein, the ulinastatin that the present embodiment obtains is white powder, and adopting and recording purity with the method for embodiment 1 is 99.7%, and the yield of ulinastatin is 67%, and Rate activity is 4010IU/mg albumen.
Buffer solution used by balance cation exchanger resin is sodium phosphate buffer system.
Above-described embodiment is only used as the explanation purpose of the present invention, and the scope of the present invention is not limited.The amendment made to one skilled in the art is apparent from, and the present invention is limited only by the restriction of scope.
Claims (6)
1., based on a method for cation exchange resin purification ulinastatin, it specifically comprises the following steps that
1) take clarification urine, regulate pH to 6-6.5, add silica gel, stirring, filter, obtain filtrate, after regulating filtrate pH to 3-5, add chitosan and alginate, stirring, obtain mixed liquor 1, be 5 × 10 by mixed liquor 1 with molecular cut off4-7×104Ultrafilter membrane 1 ultrafiltration obtain trapped fluid 1；
2) regulate the pH to 7-8 of trapped fluid 1, filter, obtain filtrate 1, upper cation exchange resin column after filtrate 1 is adjusted pH to 4-8, collect effluent and obtain effluent A；
3) it is 3 × 10 by effluent A molecular cut off4Ultrafilter membrane 2 ultrafiltration obtain trapped fluid 2；
4) by trapped fluid 2 lyophilization, pure ulinastatin is obtained.
2. method according to claim 1, wherein, described ultrafilter membrane is the composite membrane of a kind of or at least two in poly (ether sulfone) film, polypropylene screen, regenerated cellulose film, cellulose acetate membrane, polysulfone membrane and polyvinylidene fluoride film；The structure of described ultrafilter membrane is the one in hollow fiber form, flat, tubular type and rolling.
3. method according to claim 2, wherein, described ultrafilter membrane is the composite membrane of poly (ether sulfone) film and cellulose acetate membrane；The structure of described ultrafilter membrane is inner pressed hollow fiber ultrafiltration membrane.
4. the method according to any one of claim 1-3, wherein, described cation exchange resin is a kind of mixing in AmberliteIR120 and Amberlite200, AmberliteIRA118, AmberliteIRC84, Amberjet1200 and AmberliteIRC54.
5. the method according to claim 4, wherein, described cation exchange resin be the one in AmberliteIR120 and Amberlite200, AmberliteIRA118, AmberliteIRC84, Amberjet1200 and AmberliteIRC54 with mass ratio for (1-5): 1 mixing.
6. method according to claim 5, wherein, step 2) in balance the solution of described cation exchange resin column be phosphate buffer or Tris-HCl buffer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610323431.0A CN105753976A (en) | 2016-05-13 | 2016-05-13 | Method for purifying ulinastatin based on cation exchange resin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610323431.0A CN105753976A (en) | 2016-05-13 | 2016-05-13 | Method for purifying ulinastatin based on cation exchange resin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105753976A true CN105753976A (en) | 2016-07-13 |
Family
ID=56323192
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610323431.0A Pending CN105753976A (en) | 2016-05-13 | 2016-05-13 | Method for purifying ulinastatin based on cation exchange resin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105753976A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107827977A (en) * | 2017-11-15 | 2018-03-23 | 广东天普生化医药股份有限公司 | A kind of method based on ion-exchange resin purification UTI |
| CN107936114A (en) * | 2017-11-08 | 2018-04-20 | 广东天普生化医药股份有限公司 | A kind of method based on cation exchange resin purifying ulinastatin |
| CN108059670A (en) * | 2017-11-01 | 2018-05-22 | 广东天普生化医药股份有限公司 | A kind of ulinastatin purification process based on affinity column |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105238771A (en) * | 2015-10-23 | 2016-01-13 | 张昭 | Method for extracting urokinase and ulinastatin from urine |
| CN105384813A (en) * | 2015-11-24 | 2016-03-09 | 青岛康原药业有限公司 | Method for purifying ulinastatin by adsorption column chromatography and medicine composition for improving stability of ulinastatin |
-
2016
- 2016-05-13 CN CN201610323431.0A patent/CN105753976A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105238771A (en) * | 2015-10-23 | 2016-01-13 | 张昭 | Method for extracting urokinase and ulinastatin from urine |
| CN105384813A (en) * | 2015-11-24 | 2016-03-09 | 青岛康原药业有限公司 | Method for purifying ulinastatin by adsorption column chromatography and medicine composition for improving stability of ulinastatin |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108059670A (en) * | 2017-11-01 | 2018-05-22 | 广东天普生化医药股份有限公司 | A kind of ulinastatin purification process based on affinity column |
| CN107936114A (en) * | 2017-11-08 | 2018-04-20 | 广东天普生化医药股份有限公司 | A kind of method based on cation exchange resin purifying ulinastatin |
| CN107936114B (en) * | 2017-11-08 | 2019-01-22 | 广东天普生化医药股份有限公司 | A method of ulinastatin is purified based on cation exchange resin |
| CN107827977A (en) * | 2017-11-15 | 2018-03-23 | 广东天普生化医药股份有限公司 | A kind of method based on ion-exchange resin purification UTI |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105753976A (en) | Method for purifying ulinastatin based on cation exchange resin | |
| CN103232558B (en) | Preparation method of high-quality low-molecular weight dalteparin sodium | |
| CN109925742B (en) | Chromatographic separation device and method for separating vitamin C mother liquor by using same | |
| CN105154499B (en) | The preparation method of L-aspartic acid-L-ornithine | |
| CN108358989A (en) | A method of isolating and purifying cytidine from microbial fermentation solution | |
| JPH0459878B2 (en) | ||
| CN109503705A (en) | A kind of isolation and purification method of Liraglutide | |
| CN115925890A (en) | Method for purifying anti-new coronavirus neutralizing antibody | |
| CN102603814B (en) | Method for increasing crystalizing efficiency of xylose in xylose mother solution | |
| CN109553650B (en) | Water phase extraction method of erythromycin fermentation liquor | |
| CN114181300A (en) | Preparation method of high-purity monoclonal antibody | |
| CN108822164A (en) | The preparation process of high quality monosialotetrahexose ganglioside sodium | |
| WO2020147421A1 (en) | Sugammadex isolation and purification method | |
| CN107827977A (en) | A kind of method based on ion-exchange resin purification UTI | |
| CN105753977A (en) | Method for large-scale production of high-purity ulinastatin (namely human urinary trypsin inhibitor) | |
| CN110451529A (en) | A kind of method of purification of sodium chloride for injection | |
| CN105837685A (en) | Method for purifying ulinastatin based on anion exchange resin | |
| CN104313071B (en) | The biological synthesis method of high-purity L alpha amino acids | |
| CN102584611B (en) | Production method for medical grade valine | |
| CN103012506A (en) | Energy-saving process for extracting crystallized xylose and arabinose from xylose mother liquor | |
| CN105255852B (en) | Utilize the method for ceramic membrane separation immobilised enzymes and heparin enzymolysis liquid | |
| CN220788618U (en) | D-chiro-inositol's apparatus for producing | |
| CN100540521C (en) | A kind of gulonic acid mother solution of using reclaims the sour practical art of ancient dragon | |
| CN112760309B (en) | Method for multi-stage purification of nuclease P1 | |
| CN106554374A (en) | A kind of method that purification prepares vistamycin in fermentation liquid from ribostamycin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160713 |