CN105622747A - vWF (von Willebrand factor) activity protection fluid - Google Patents
vWF (von Willebrand factor) activity protection fluid Download PDFInfo
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- CN105622747A CN105622747A CN201610077438.9A CN201610077438A CN105622747A CN 105622747 A CN105622747 A CN 105622747A CN 201610077438 A CN201610077438 A CN 201610077438A CN 105622747 A CN105622747 A CN 105622747A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/745—Blood coagulation or fibrinolysis factors
- C07K14/755—Factors VIII, e.g. factor VIII C (AHF), factor VIII Ag (VWF)
Abstract
The invention discloses a vWF (von Willebrand factor) activity protection fluid. The vWF activity protection fluid is used in preparation for extracting von Willebrand factors from cryoprecipitated blood coagulation factor VIII waste. In a preparation technology of the von Willebrand factors, glycine is added into a chromatographic buffer solution, and lysine, glycine and albumin are added into protein fluid before freeze drying after chromatography to protect vWF activity. The vWF activity protection fluid has the advantages that by means of adding the glycine into the chromatographic buffer solution, vWF activity loss can be reduced in a chromatographic separation and purification process; the albumin, the lysine and the glycine are added into protein dialysate to serve as freeze-drying protectants, so that von Willebrand factor molecules can be stabilized; the albumin serving as an excellent protein stabilizer is capable of effectively adsorbing protein surfaces; the lysine and the glycine, serving as micromolecular amino acids, are capable of protecting a protein structure, increasing collapse temperature of a finished product and stopping protein damage caused by collapse during freeze-drying, so that biological activity is kept.
Description
Technical field
The present invention relates to the preparation technology extracting human von willebrand disease factor a kind of waste material extracting platelet cofactor �� from human plasma cryoprecipitate, particularly to a kind of vWF active protection liquid, belong to field of biological pharmacy.
Background technology
VWF is that one has polymeric plasma glycoprotein, and its molecular weight from 250kDa to 2,000 ten thousand kDa not etc., forms maximum molecular weight soluble protein known in blood plasma. Blood plasma small molecular amount vWF is mainly dimeric forms, and molecular weight is about 500kDa. The polymer differed in size, to maintaining, vWF normal biological activity is significant.
Plasma vWF plays an important role in constitutional stops blooding, and it is responsible for the adhesion of platelet and damaged blood vessel surface, and therefore forms platelet plug, contributes to the formation of fibrin crosslinking. It addition, plasma vWF plays a part transport agent and the stabilizer of blood coagulation factor VIII.
Von Willebrand (vWD) is modal hereditary hemorrhagic disease, and (vonWillebrandfactor, the vWF) gene mutation of patient's vWF or regulation and control cause that plasma vWF quantity reduces or abnormal quality extremely. It is hemorrhage that vWD Clinical symptoms is mainly skin mucosa, and 2N type and 3 type patients also can occur the articular cavity similar to hemophilia A hemorrhage and muscle hematoma. According to external, vWD sickness rate (2.3��11.0)/100,000 population, it is considered that in hereditary hemorrhagic disease, vWD occupies second.
The activity Ristocetin co-factor activity (vWF:RCo) of vWF characterizes clinically, is mainly used in treating congenital, Of Acquired von Willebrand Disease (vWD), is generally administered with 20��60UvWF:RCo/kg dosage.
Research shows in cryoprecipitate rich in blood clotting factors materials such as vWF. In today that blood plasma raw material is increasingly in short supply, prepare vWF by cryoprecipitate, the comprehensive utilization to cryoprecipitate, save rare blood plasma resource, improve the market competitiveness and be significant.
In prior art, technique uses the antibody coupling of vWF to agarose gel earlier, by immunoaffinity chromatography purification vWF, but its antibody manufacturing cycle is long, gel carrying capacity is low, the extensive preparation of uncomfortable cooperation, such as " PurificationofvonWillebrandFactorsolutionsusinggelpermea tionchromatography " (UP4774323).
There is patent DEAEFractogelTSK650M (Merck) purification vWF factor component, purity is high, but technics comparing is complicated, need two-step solution and a step affinity chromatograph, such as " Processforanindustrial-scalepreparationofastandardizedhu manvonWillebrandfactorconcentrateofveryhighpurityandsuit ablefortherapeuticuse " (US5408039). Also have been reported that CHT chromatography separates vWF component in conjunction with PH5.4 Acid precipitation, it is not higher than living, and while Acid precipitation removal Fn Fiberonectin, it is likely to affect the biologic activity of vWF, such as " Productionofavonwillebrandfactorprepartionhavingagreatsp ecificactivity " (US20070135619A1).
Some technique EMDFractogelTMAE (Merck) are additionally also had to carry out purification VIII/vWF factor coniplexes, but all cannot individually obtain highly purified vWF, treatment specific aim is not strong, such as " preparation containing vWF with styptic activity and preparation method thereof " (CN1425024A), " AProcessforrecoveringahigh-purityvirus-inactivatedfactor VIIIbyanionexchangerchromatography " (US5714590).
Also has method (monkey-kidney cells or Chinese hamster ovary celI) the purification vWF of some company's gene recombinaton, they are purified from animal cell culture fluid by anion exchange chromatography, but the existence due to animal originality antigen, it is likely to there is repulsive interaction by human body, such as " MethodforisolationofhighlypurevonWillebrandFactor " (US5854403).
Separately having the report adopting two-step chromatography purification vWF, but its purity is not high, more relatively low than living, only 50IU/mg, such as " ProcessformanufacturingvonWillebrandfactor " (US5252710).
Visible, existing vWF purifying process still suffers from some problems: (1) manufacturing cycle is long, is not suitable for extensive preparation, and during in particular by immune-affinity chromatography purification, its antibody manufacturing cycle is long, and gel carrying capacity is low. (2) VIII/vWF composite product is at more difficult control technology stability; (3) recombination method gained vWF product would be likely to occur heterogeneic antigen; (4) separation purifying technique is loaded down with trivial details.
At present, von Willebrand can use cryoprecipitate, VIII/vWF factor coniplexes and DDAVP (DDAVP) treatment, but specific aim is not strong, the purer vWF factor can the various types of von Willebrand of specific therapeutic, have greater advantage with front several method ratio.
Summary of the invention
It is an object of the invention to provide a kind of vWF active protection liquid.
The major technique design of the present invention is as follows:
Protein liquid after chromatography is added glycine protection vWF activity by the present invention in chromatography buffer; Protein liquid before lyophilizing adds lysine, glycine and albumin as freeze drying protectant, protection vWF activity after preferred chromatography; Glycine mass concentration 0.6%��1% in preferred chromatography buffer, lysine mass concentration 2��3% in freeze drying protectant, glycine mass concentration 0.2%, albumin mass concentration 0.8��1.3%.
Described protection liquid is divided into lavation buffer solution B, elution buffer B and dialysis solution, when carrying out chromatography purification with Q-Sepharose anion-exchange chromatography post, washs pillar with lavation buffer solution B until becoming baseline, with elution buffer B eluting; When gelatin affinity chromatograph is purified, wash pillar with elution buffer B until becoming baseline, the liquid come through chromatographic column effluent after collecting loading, dialyse with dialysis solution.
The formula of lavation buffer solution B: 4g sodium citrate, 11.4g sodium chloride, 3g calcium chloride, 6g glycine, add appropriate water for injection and fully dissolve, mend and inject water to 1L, regulating PH is 5.8��6.8.
Elution buffer B formula: 4g sodium citrate, 16.38g sodium chloride, 3g calcium chloride, 6g glycine, add appropriate water for injection and fully dissolve, mend and inject water to 1L, regulating PH is 5.8��6.8.
The formula of dialysis solution: 2.94g sodium citrate, 25g sodium chloride, 0.111g calcium chloride, 30g glycine, 2g lysine hydrochloride, human albumin 8g, add appropriate water for injection and fully dissolve, mend and inject water to 1L, regulating PH is 6.8��7.2.
Extract the preparation technology extracting vWF ELISA the waste material of platelet cofactor �� from cryoprecipitate, it includes successively: cryoprecipitate is dissolved, 2% gel aluminum hydroxide adsorbs, regulate ionic strength, cascade filtration, S/D inactivation of virus, ion-exchange chromatography; Ion-exchange chromatography must collect eluent, and namely eluent, through ultrafiltration, dialysis, filtration, for the production of blood coagulation factor VIII, then collects extraction vWF through the liquid that chromatographic column stream passes from the waste material of cryoprecipitate extraction platelet cofactor ��; Q-Sepharose anion-exchange chromatography, gelatin affinity chromatograph, aseptic filtration subpackage, lyophilization, xeothermic inactivation. After ion-exchange chromatography stream wears liquid collection, first concentration one times, then with lavation buffer solution B equal-volume ultrafiltration 6 times, then through Q-Sepharose chromatographic column, collect the protein liquid eluted with elution buffer B.
The concrete technology scheme extracting vWF ELISA the waste material of platelet cofactor �� is extracted as follows from cryoprecipitate:
(1) after qualified human plasma of quarantining quanrantine is got, 75% ethanol plasma bags surface, to rinse with water for injection, be merged into and melt in slurry tank, melt with less than 30��35 DEG C recirculated waters, blood plasma temperature controls not higher than 4 DEG C; After melting, centrifugal, go out liquid temp and control at 0��4 DEG C, collect cryoprecipitate;
(2) adding in 3IU/ml heparin sodium aqua by the prepared thing of step (1), stirring is completely dissolved to cryoprecipitate, and the temperature of recirculated water controls at 28��37 DEG C; Start to be centrifuged, collect supernatant, weigh;
(3) the prepared thing supernatant 1mol/L hydrochloric acid of step (2) is regulated PH to 6.0��7.0; Add 2% gel aluminum hydroxide, stirring; Start to be centrifuged, collect supernatant, weigh;
(4) calculate by " W (ionic strength buffer liquid)=supernatant weight/10 ", weigh the ionic strength buffer liquid of amount of calculation to the prepared thing supernatant of step (3); Regulate supernatant PH to 6.0��7.0; Regulate protein concentration with protein concentration buffer and be not more than 10g/L; With the filter element filtering of 1.0 ��m, collect filtrate, weigh;
(5) adding S/D solution by the 1/10 of the prepared thing filtrate volume of step (4), stir, temperature controls at 24��26 DEG C, continuously insulation 6 hours; 0.45 ��m of filter element filtering; Filtrate 30KD ultrafilter membrane is concentrated into protein concentration 2%, then with more than 2 times lavation buffer solution A constant volume ultrafiltration of protein liquid weight, obtains ultrafiltrate, weighs;
(6) the prepared thing ultrafiltrate ion exchange column of step (5) is carried out chromatography purification, pillar is washed until becoming baseline with lavation buffer solution A, with elution buffer A eluting, collect eluent (eluent through ultrafiltration, dialysis, filtration, for the production of blood coagulation factor VIII).
(7) collect the liquid that step (6) is come through chromatographic column effluent, concentrate one times, lavation buffer solution B equal-volume ultrafiltration 6 times, obtain ultrafiltrate, weigh.
(8) the prepared thing ultrafiltrate Q-Sepharose anion-exchange chromatography post of step (7) being carried out chromatography purification, washing pillar with lavation buffer solution B until becoming baseline, with elution buffer B eluting, collect eluent, weigh.
(9) the prepared thing eluent gelatin affinity chromatograph of step (8) is purified, pillar is washed until becoming baseline with elution buffer B, the liquid come through chromatographic column effluent after collecting loading, dialyse with dialysis solution, be diluted to titer 100IU/ml, obtain protein liquid, weigh.
(10) by the prepared thing protein liquid of step (9) through 0.2 ��m of degerming filter element filtering subpackage; The incoming lyophilizing cabinet of goods that subpackage is good carries out lyophilization; Offer for sale Zha Gai; 99��100 DEG C, 30 minutes xeothermic inactivation of virus; Censorship, packs after the assay was approved, puts in storage.
The compound method of ionic strength buffer liquid: 48.5g trishydroxymethylaminomethane, 10g calcium chloride, 100g sodium chloride, 28.1g glycine, add appropriate water for injection and fully dissolve, mend and inject water to 1L, regulating PH is 6.0��7.0; The compound method of protein concentration buffer: measure 0.05L and regulate ionic strength buffer liquid, inject water to 1L, regulating PH is 6.0��7.0.
Lavation buffer solution A formula: 2.5g trishydroxymethylaminomethane, 10g calcium chloride, 25g sodium chloride, add appropriate water for injection and fully dissolve, mend and inject water to 1L, regulating PH is 6.0��7.0.
Elution buffer A formula: 2.5g trishydroxymethylaminomethane, 15g calcium chloride, 40.9g sodium chloride, add appropriate water for injection and fully dissolve, mend and inject water to 1L, regulating PH is 6.0��7.0.
Described percent is except having restriction, and all the other are mass percent.
The good effect extracting vWF ELISA the waste material of platelet cofactor �� is extracted from cryoprecipitate:
1, in today that blood plasma raw material is increasingly in short supply, make full use of blood plasma resource, wear waste liquid with the stream of blood coagulation factor VIII ion exchange and prepare high-purity vWF. Waste material such as vWF, Fibrinogen etc. can be produced when producing blood coagulation factor VIII, by the Component seperation in waste material out, turn waste into wealth, utilize blood plasma resource in short supply to greatest extent, improve blood plasma utilization rate.
2, adopt the exchange of step ion and a step affinity chromatograph technique, both can guarantee that the purity of vWF and yield, it is possible to simplify production technology, fully reduce manpower and time cost. Ion-exchange chromatography plays the effect of preliminary purification, and affinity chromatograph filler has the feature of binding proteins specific, further albumen is carried out polishing purification.
3, adopt the cryoprecipitate of humanized as initiation material, it is to avoid the generation of the heterogeneic antigen rejection of recombiant protein, improve Clinical practice safety.
The present invention preparation technology compared to tradition vWF, innovative point is in that:
1, chromatography buffer is added glycine, protein liquid before lyophilizing adds after chromatography lysine, glycine and albumin, protection vWF activity. Chromatography buffer adds glycine the loss of activity of vWF in chromatography purge process, can be reduced. Albumen dialysis solution adds albumin, lysine, glycine as freeze drying protectant, vWF molecule can be stablized. Albumin is excellent protein stabiliser, simultaneously can the absorption of effective protein surface. Lysine, glycine are little molecule aminoacid, it is possible to protected protein matter structure, can raise the collapse temperature of finished product, stop the Protein Damage caused in freeze-drying process because subsiding. Keep biologic activity.
3, ion-exchange chromatography separates the salt ionic concentration that the most critical factor of vWF is cleaning mixture and eluent. The too low meeting of cleaning mixture salinity causes the impurity such as Fibrinogen, Fn Fiberonectin remaining, and cleaning mixture salinity is too high, the response rate causing vWF is reduced, and the vWF amount afforded reduces. Finding after test of many times, 195mM (11.4g/L) sodium chloride is best cleaning mixture salinity. Additionally eluent salt concentration also need in optimum range, and the too high too low acquisition being also unfavorable for vWF, we are defined as 280mM (16.38g/L) by test. The proportioning of both salinity can take into account the activity of vWF and than living, and is best a pair matched proportion density.
Accompanying drawing explanation
Fig. 1 is the embodiment of the present invention 1 process chart.
It is embodied as case
The present invention by the following waste material extracting platelet cofactor �� from cryoprecipitate extracts vWF ELISA embodiment can the invention will be further described, but, the scope of the present invention is not limited to following embodiment.
Embodiment 1
For 6000L blood plasma, concrete preparation technology is as follows:
(1) after qualified human plasma of quarantining quanrantine is got, 75% ethanol plasma bags surface, to rinse with water for injection, be merged into and melt in slurry tank, melt with less than 30��35 DEG C recirculated waters, blood plasma temperature controls not higher than 4 DEG C; After melting, centrifugal, go out liquid temp and control, at 0��4 DEG C, to collect to obtain cryoprecipitate 45.2kg;
(2) adding in 3IU/ml heparin sodium aqua by the prepared thing cryoprecipitate of step (1), stirring is completely dissolved to cryoprecipitate, and the temperature of recirculated water controls at 28��37 DEG C; Starting to be centrifuged, collect supernatant, weigh to obtain 169.5kg;
(3) the prepared thing supernatant 1mol/LHCL of step (2) is regulated PH to 6.0��7.0; Add 2% gel aluminum hydroxide, stirring; Starting to be centrifuged, collect supernatant, weigh to obtain 178.6kg;
(4) calculate by " W (ionic strength buffer liquid)=supernatant weight/10 ", weigh the ionic strength buffer liquid of amount of calculation to the prepared thing supernatant of step (3); Regulate supernatant PH to 6.0��7.0; Regulate protein concentration with protein concentration buffer and be not more than 10g/L; Filtering after connecting with the filter element of the filter elements of 1.0 ��m and 0.45 ��m, collect filtrate, weigh to obtain 189.9kg;
(5) adding S/D solution by the 1/10 of the prepared thing filtrate volume of step (4), stir, temperature controls at 24��26 DEG C, continuously insulation 6 hours; 0.45 ��m of filter element filtering; Filtrate 30KD ultrafilter membrane is concentrated into protein concentration about 2%, and then with more than 2 times lavation buffer solution A constant volume ultrafiltration of protein liquid weight, i.e. ultrafiltrate, weigh to obtain 201.4L;
(6) the prepared thing ultrafiltrate ion exchange column of step (5) being carried out chromatography purification, washing pillar with lavation buffer solution A until becoming baseline, with elution buffer A eluting, collect eluent.
(7) collect the liquid that step (6) is come through chromatographic column effluent, concentrate one times, lavation buffer solution B equal-volume ultrafiltration 6 times, obtain ultrafiltrate, weigh to obtain 120.5L;
(8) the prepared thing ultrafiltrate Q-Sepharose anion-exchange chromatography post of step (7) being carried out chromatography purification, washing pillar with lavation buffer solution B until becoming baseline, with elution buffer B eluting, collect eluent, weigh to obtain 48.5L;
(9) the prepared thing eluent gelatin affinity chromatograph of step (8) being purified, washing pillar with elution buffer B until becoming baseline, the liquid come through chromatographic column effluent after collecting loading, it is concentrated by ultrafiltration to 5L, dialysing 6 times, obtain ultrafiltrate, weigh to obtain 5.2L; It is assigned to 6.1L by titer 100IU/ml is rare;
(10) by the prepared thing filtrate of step (9) through 0.2 ��m of degerming filter element filtering subpackage, subpackage loading amount is every bottle of 5mL, and subpackage quantity is 1196 bottles; The incoming lyophilizing cabinet of goods that subpackage is good carries out lyophilization; Offer for sale Zha Gai; 99��100 DEG C, 30 minutes xeothermic inactivation of virus; Censorship, packs after the assay was approved, puts in storage;
Described percent is except having restriction, and all the other are mass percent.
In product prepared by the inventive method, the lab scale of the impact of product quality is studied by different ion exchange salinity, such as table 1 below.
Table 1:
Illustrate: in anion exchange procedures, vWF and Fn Fiberonectin are interval in this salinity of 150mMNaCl to 300mMNaCl, salinity and ionic strength is very sensitive, through Experimental Comparison analysis, screening obtains the column buffer formula of liquid that two kinds of major impurities (Fn Fiberonectin and Fibrinogen) are minimum. Preferred cleaning mixture is 195mM (11.4g/L) NaCl and eluent is 280mM (16.38g/L) NaCl, can obtain activity and than all higher vWF that lives.
In product prepared by the inventive method, the glycine impact on product in chromatography buffer, such as table 2.
Table 2:
| Glycine concentration (%) | Cleaning mixture belit concentration (%) | Eluent belit concentration (%) | VWF:RCo activity (IU/ml) |
| 0 | 195 | 280 | 101.5 |
| 0.2 | 195 | 280 | 102.1 |
| 0.6 | 195 | 280 | 104.8 |
| 1.0 | 195 | 280 | 104.81 |
Illustrate: the glycine in chromatography buffer in chromatography process, can protect the effect in vWF, mass concentration 0.6%��1%, and the protective effect of chromatography process vWF is more or less the same by the glycine concentration of this scope, select relatively low, it is still further preferred that 0.6%.
In product prepared by the inventive method, the impact on product of dialysis solution (frozen-dried protective liquid) the various heterogeneity, such as table 3.
Table 3:
Illustrating: glycine mass concentration 2��3%, lysine mass concentration 0.2%, albumin mass concentration 0.8��1.3%, for the good scope of application, outward appearance is slightly or without atrophy, and has slight or without opalescence, activity is also better protected. The difference of the mass concentration of lysine is less on frozen-dried protective impact, selects 0.2%. Best dialysis buffer liquid (frozen-dried protective liquid) formula is glycine concentration is 3%, and albumin concentration is 0.8%, and outward appearance and activity are the best.
The contrast such as table 4 below of the vWF Key Quality Indicator that the embodiment product extracting vWF ELISA the waste material of platelet cofactor �� describes is extracted with " in European Pharmacopoeia " from cryoprecipitate.
Table 4:
| Project | The present invention | " European Pharmacopoeia " |
| PH | 6.8��7.2 | 6.5��7.5 |
| Activity | Should >=100IU/ml | ��1IU/ml |
| Than living | Should >=200IU/mg | ��20IU/mg |
| Blood coagulation factor VIII content | VIII:C 2��4IU/ml in every 100IU vWF:RCo | VIII:C��10IU/ml in every 100IUvWF:RCo |
| Osmotic pressure molar density | 240��1000mOsmol/kg | 240mOsmol/kg should be not less than |
Claims (6)
1. a vWF active protection liquid; for extracting from cryoprecipitate, the waste material of platelet cofactor �� extract in the preparation of vWF ELISA; it is characterized in that; in the preparation technology of vWF ELISA; chromatography buffer is added glycine; protein liquid before lyophilizing adds after chromatography lysine, glycine and albumin, protection vWF activity.
2. a kind of vWF active protection liquid according to claim 1; it is characterized in that, glycine mass concentration 0.6%��1% in chromatography buffer, lysine mass concentration 2��3% in freeze drying protectant; glycine mass concentration 0.2%, albumin mass concentration 0.8��1.3%.
3. a kind of vWF active protection liquid according to claim 1, it is characterized in that, described protection liquid is divided into lavation buffer solution B, elution buffer B and dialysis solution, when carrying out chromatography purification with Q-Sepharose anion-exchange chromatography post, pillar is washed until becoming baseline, with elution buffer B eluting with lavation buffer solution B; When gelatin affinity chromatograph is purified, wash pillar with elution buffer B until becoming baseline, the liquid come through chromatographic column effluent after collecting loading, dialyse with dialysis solution.
4. a kind of vWF active protection liquid according to claim 1 and 2, it is characterised in that the formula of described lavation buffer solution B: 4g sodium citrate; 11.4g sodium chloride, 3g calcium chloride, 6g glycine; adding appropriate water for injection fully to dissolve, mend and inject water to 1L, regulating pH is 5.8��6.8.
5. a kind of vWF active protection liquid according to claim 1 and 2, it is characterised in that described elution buffer B formula: 4g sodium citrate; 16.38g sodium chloride, 3g calcium chloride, 6g glycine; adding appropriate water for injection fully to dissolve, mend and inject water to 1L, regulating pH is 5.8��6.8.
6. a kind of vWF active protection liquid according to claim 1 and 2; it is characterized in that; described dialysis solution formula: 2.94g sodium citrate, 25g sodium chloride, 0.111g calcium chloride; 30g glycine; 2g lysine hydrochloride, human albumin 8g, add appropriate water for injection and fully dissolve; benefit injects water to 1L, and regulating pH is 6.8��7.2.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018102960A1 (en) * | 2016-12-05 | 2018-06-14 | Chung Chin Sun | Novel method for cryoprecipitate activity preservation |
| CN110997015A (en) * | 2017-08-23 | 2020-04-10 | 德国杰特贝林生物制品有限公司 | Methods for viral filtration of von Willebrand factor |
| TWI723095B (en) * | 2016-12-05 | 2021-04-01 | 英屬維爾京群島商大利緯眾生物科技股份有限公司 | A novel method for cryoprecipitate activity preservation |
| CN112851795A (en) * | 2019-11-27 | 2021-05-28 | 菲鹏生物股份有限公司 | Stabilizer, preservation reagent, kit and preservation method for protein and polypeptide |
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| CA2009946C (en) * | 1989-02-14 | 2000-04-11 | Norbert Heimburger | Pasteurized, purified von willebrand factor concentrate and a process for the preparation thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018102960A1 (en) * | 2016-12-05 | 2018-06-14 | Chung Chin Sun | Novel method for cryoprecipitate activity preservation |
| TWI723095B (en) * | 2016-12-05 | 2021-04-01 | 英屬維爾京群島商大利緯眾生物科技股份有限公司 | A novel method for cryoprecipitate activity preservation |
| CN110997015A (en) * | 2017-08-23 | 2020-04-10 | 德国杰特贝林生物制品有限公司 | Methods for viral filtration of von Willebrand factor |
| CN112851795A (en) * | 2019-11-27 | 2021-05-28 | 菲鹏生物股份有限公司 | Stabilizer, preservation reagent, kit and preservation method for protein and polypeptide |
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Inventor after: Yang Ducai Inventor after: Liang Xiaoming Inventor after: Yang Zhi Inventor after: Mai Xi Inventor after: Zhen Xie Inventor after: Zhang Meng Inventor before: Yang Ducai Inventor before: Liang Xiaoming Inventor before: He Shuqin Inventor before: Yang Zhi Inventor before: Liu Yuliang Inventor before: Kuang Qingfen |
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Application publication date: 20160601 |