CN1324048C - Method of eliminating endotoxin from interferon preparation - Google Patents

Method of eliminating endotoxin from interferon preparation Download PDF

Info

Publication number
CN1324048C
CN1324048C CNB031110363A CN03111036A CN1324048C CN 1324048 C CN1324048 C CN 1324048C CN B031110363 A CNB031110363 A CN B031110363A CN 03111036 A CN03111036 A CN 03111036A CN 1324048 C CN1324048 C CN 1324048C
Authority
CN
China
Prior art keywords
interferon
endotoxin
affinity
remove
chitosan
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.)
Expired - Lifetime
Application number
CNB031110363A
Other languages
Chinese (zh)
Other versions
CN1523037A (en
Inventor
李京华
邵英光
丛润滋
王俊德
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Beijing Furui Tiancheng Biotechnology Co ltd
Beijing Kawin Technology Share Holding Co ltd
Original Assignee
Dalian Institute of Chemical Physics of CAS
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Dalian Institute of Chemical Physics of CAS filed Critical Dalian Institute of Chemical Physics of CAS
Priority to CNB031110363A priority Critical patent/CN1324048C/en
Publication of CN1523037A publication Critical patent/CN1523037A/en
Application granted granted Critical
Publication of CN1324048C publication Critical patent/CN1324048C/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Landscapes

  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)

Abstract

本发明涉及重组蛋白质药物的生产,具体地说是一种从干扰素制剂中去除内毒素的方法,将壳聚糖,或去氧胆酸钠,或己二胺作为亲和配基,共价结合到多孔介质上,以动态过滤或静态吸附的方式进行操作,用以去除重组干扰素制剂中的内毒素;其中亲和配基在多孔介质上的量为含氮量0.02%~3%。本发明方法内毒素去除率高,干扰素活性回收率高;特异性好,适合生产的要求;实用范围广,前景好。The present invention relates to the production of recombinant protein medicine, specifically a method for removing endotoxin from interferon preparations, using chitosan, or sodium deoxycholate, or hexamethylenediamine as an affinity ligand, covalently Combined with porous media, it is operated by dynamic filtration or static adsorption to remove endotoxin in recombinant interferon preparations; the amount of affinity ligand on porous media is 0.02% to 3% nitrogen content. The method of the invention has high removal rate of endotoxin and high recovery rate of interferon activity; good specificity, suitable for production requirements; wide practical range and good prospect.

Description

一种从干扰素制剂中去除内毒素的方法A method for removing endotoxin from interferon preparations

技术领域technical field

本发明涉及重组蛋白质药物的生产,具体地说是一种从干扰素制剂中去除内毒素的方法。The invention relates to the production of recombinant protein medicine, in particular to a method for removing endotoxin from interferon preparations.

背景技术Background technique

随着基因工程技术的日益成熟,越来越多的重组蛋白质药物被开发利用,由于生产过程中涉及到革兰氏阴性菌及分离纯化过程复杂,因此会造成内毒素的污染。With the increasing maturity of genetic engineering technology, more and more recombinant protein drugs have been developed and utilized. Since the production process involves Gram-negative bacteria and the separation and purification process is complicated, it will cause endotoxin pollution.

亲和技术是分离纯化生物大分子的有效手段,主要利用配基与生物大分子之间的亲和作用原理;将配基通过共价键键合到多孔介质上就得到亲和介质,可以用来选择性地吸附内毒素,实现从重组蛋白质溶液中去除内毒素的目标;已有的亲和配基和亲和介质包括:单克隆抗体,多克隆抗体,抗生素,多粘菌素B,聚乙烯亚胺,组氨酸,凝胶过滤柱,亲和膜,颗粒吸附剂等。Affinity technology is an effective means of separating and purifying biomacromolecules, which mainly utilizes the principle of affinity between ligands and biomacromolecules; the ligands are covalently bonded to porous media to obtain affinity media, which can be used To selectively adsorb endotoxins and achieve the goal of removing endotoxins from recombinant protein solutions; existing affinity ligands and affinity media include: monoclonal antibodies, polyclonal antibodies, antibiotics, polymyxin B, polyethylene Imine, histidine, gel filtration columns, affinity membranes, particle adsorbents, etc.

由于重组蛋白质种类较多,其分离纯化步骤各不相同,生产过程中对其中的内毒素含量要求也有差异,因此去除内毒素的方法和效果无法完全通用。韩国专利(KR8902068)采用凝胶过滤柱Sephacryl S-200,利用分子筛原理来分离分子大小不同的内毒素和干扰素,以除去干扰素中的内毒素;但流速低,为4ml/hr,过程中需要添加其它物质如Triton X-100。欧洲专利(EP0800862)采用含有磺酸基的苯乙烯-二乙烯基苯共聚物,利用离子交换的原理,来去除重组蛋白质肿瘤坏死因子或白介素中的内毒素,主要吸附重组蛋白质。欧洲专利(EP0337243)使用串联的两只反相液相色谱柱,来纯化重组人IL-2,在纯化重组人白介素-2的同时,去除内毒素杂质,流速低,介质昂贵。美国专利(US4885168)采用低分子量壳聚糖,利用壳聚糖上带正电荷的氨基来与带负电荷的核酸与内毒素作用,达到除去核酸与内毒素的目的,但壳聚糖的利用率低。中国专利(00123290.6)魏桂林等的一种用于内毒素去除的疏水荷正电亲和膜,将胺类疏水荷正电亲和配基键合到纤维素膜上,得到相应的亲和膜;胺类疏水荷正电亲和配基包括2~12碳的烃基二胺、带有乙烯基的不饱和杂环化合物、带有环氧基团的季铵盐及带有多个疏水基团的季铵盐;其主要应用于水中内毒素的去除。D.Petsch,T.C.Beeskow等(1997,Journal of Chromatography B.693,79-91)采用去氧胆酸钠,或己二胺作为亲和配基,以微孔尼龙膜为介质,制成亲和膜;采用单张亲和膜,可以去除缓冲液或牛血清白蛋白中的内毒素;但流速低,阻力高,处理量少,不易放大。Since there are many types of recombinant proteins, their separation and purification steps are different, and the requirements for the endotoxin content in the production process are also different, so the method and effect of removing endotoxin cannot be completely universal. Korean patent (KR8902068) adopts gel filtration column Sephacryl S-200, uses the principle of molecular sieve to separate endotoxin and interferon with different molecular sizes, to remove endotoxin in interferon; but the flow rate is low, 4ml/hr, during the process Need to add other substances such as Triton X-100. The European patent (EP0800862) uses styrene-divinylbenzene copolymers containing sulfonic acid groups and uses the principle of ion exchange to remove endotoxins in recombinant protein tumor necrosis factor or interleukin, mainly to adsorb recombinant proteins. The European patent (EP0337243) uses two reversed-phase liquid chromatography columns in series to purify recombinant human IL-2. While purifying recombinant human interleukin-2, endotoxin impurities are removed, the flow rate is low, and the medium is expensive. U.S. Patent (US4885168) adopts low molecular weight chitosan, and utilizes positively charged amino groups on chitosan to interact with negatively charged nucleic acid and endotoxin to achieve the purpose of removing nucleic acid and endotoxin, but the utilization rate of chitosan Low. Chinese patent (00123290.6) Wei Guilin et al. describes a hydrophobic and positively charged affinity membrane for endotoxin removal. The amine hydrophobic and positively charged affinity ligand is bonded to the cellulose membrane to obtain the corresponding affinity membrane. ; Amines hydrophobic and positively charged affinity ligands include 2-12 carbon hydrocarbon diamines, unsaturated heterocyclic compounds with vinyl groups, quaternary ammonium salts with epoxy groups, and multiple hydrophobic groups Quaternary ammonium salt; it is mainly used in the removal of endotoxins in water. D.Petsch, T.C.Beeskow et al. (1997, Journal of Chromatography B.693, 79-91) used sodium deoxycholate or hexamethylenediamine as the affinity ligand, and used the microporous nylon membrane as the medium to make an affinity Membrane; a single affinity membrane can remove endotoxin in buffer or bovine serum albumin; but the flow rate is low, the resistance is high, the processing volume is small, and it is not easy to scale up.

针对干扰素制剂生产过程中的内毒素,需要根据其分子量大小、等电点的范围、溶液中其它物质的含量和种类,来判断和选择亲和配基的种类和亲和介质,达到去除内毒素、又尽可能减少干扰素活性损失的目的。现有技术的方法并不能满足干扰素制剂生产的需求。For the endotoxin in the production process of interferon preparations, it is necessary to judge and select the type of affinity ligand and affinity medium according to its molecular weight, the range of isoelectric point, and the content and type of other substances in the solution to achieve the removal of endotoxin , and the purpose of reducing the loss of interferon activity as much as possible. The methods in the prior art cannot meet the requirements for the production of interferon preparations.

发明内容Contents of the invention

本发明的目的在于,提供一种流速较高,处理量大,符合生产要求的从干扰素制剂中去除内毒素的方法。The purpose of the present invention is to provide a method for removing endotoxin from interferon preparations with high flow rate, large processing capacity and meeting production requirements.

为实现上述目的,本发明采用的技术方案为:将壳聚糖,或去氧胆酸钠,或己二胺作为亲和配基,共价结合到多孔介质上,以动态过滤或静态吸附(或静态浸泡)的方式进行操作,用以去除重组干扰素制剂中的内毒素;其中亲和配基在多孔介质上的量为:含氮量0.02%~3%。In order to achieve the above object, the technical scheme adopted in the present invention is: with chitosan, or sodium deoxycholate, or hexamethylenediamine as the affinity ligand, covalently bound to the porous medium, with dynamic filtration or static adsorption ( or static soaking) to remove the endotoxin in the recombinant interferon preparation; wherein the amount of the affinity ligand on the porous medium is: the nitrogen content is 0.02% to 3%.

所述多孔介质是含有多羟基的多糖介质,如纤维素、葡聚糖或琼脂糖;或者是含有多羟基的合成介质,如聚乙烯醇;重组干扰素溶液的pH为pH3.5~6.0;以壳聚糖为亲和配基,壳聚糖的分子量范围为1万~100万;所述多孔介质的型式为平板膜、中空纤维膜、小球或颗粒;所述动态过滤重组干扰素溶液的流速为1~20ml/min;静态吸附的操作条件为4℃至室温,静置2~24小时。The porous medium is a polysaccharide medium containing multiple hydroxyl groups, such as cellulose, dextran or agarose; or a synthetic medium containing multiple hydroxyl groups, such as polyvinyl alcohol; the pH of the recombinant interferon solution is pH3.5-6.0; Chitosan is used as the affinity ligand, and the molecular weight of chitosan ranges from 10,000 to 1 million; the type of the porous medium is flat membrane, hollow fiber membrane, pellet or particle; the dynamic filtration of the recombinant interferon solution The flow rate is 1-20ml/min; the operating condition of static adsorption is from 4°C to room temperature, and stand for 2-24 hours.

本发明具有如下优点:The present invention has the following advantages:

1.本发明方法内毒素去除率高,干扰素活性回收率高。过滤后溶液的内毒素浓度从>80EU/ml降低到<10EU/ml,干扰素的活性(效价)回收率为90%。1. The method of the present invention has a high removal rate of endotoxin and a high recovery rate of interferon activity. The endotoxin concentration of the filtered solution is reduced from >80EU/ml to <10EU/ml, and the recovery rate of interferon activity (potency) is 90%.

2.本发明特异性好(处理的重组干扰素浓度不同,或活性不同),适合生产的要求。过滤后溶液的内毒素浓度(<10EU/ml)符合药典生产要求。2. The present invention has good specificity (different concentrations or different activities of recombinant interferons are processed), and is suitable for production requirements. The endotoxin concentration (<10EU/ml) of the filtered solution meets the production requirements of the Pharmacopoeia.

3.本发明实用范围广,前景好。可以去除干扰素制剂生产中含有的内毒素污染物,解决其产品质量问题,达到药典规定的生产要求;还可望应用于其它重组蛋白质药物的生产过程,去除生物药物中内毒素等有害物质,提高我国的药物生产水平;创造较高的经济效益。3. The present invention has a wide practical range and good prospects. It can remove endotoxin pollutants contained in the production of interferon preparations, solve its product quality problems, and meet the production requirements stipulated in the Pharmacopoeia; it is also expected to be applied to the production process of other recombinant protein drugs to remove harmful substances such as endotoxins in biological drugs, Improve the level of drug production in our country; create higher economic benefits.

总而言之,亲和膜法既结合了膜的流速高、表面积大、孔径均匀、液体与配基之间扩散半径小的优势,又结合了亲和法高选择性、高效率的优点,将亲和膜制备成相应的去除器,容易实现放大以满足药物生产的需要。因此,采用亲和膜来去除干扰素制剂中的内毒素,流量高、去除效果好、有效成分回收率高,优于柱亲和层析法。All in all, the affinity membrane method not only combines the advantages of high flow rate, large surface area, uniform pore size, and small diffusion radius between the liquid and the ligand, but also combines the advantages of high selectivity and high efficiency of the affinity method. The membrane is prepared as a corresponding remover, which can be easily scaled up to meet the needs of drug production. Therefore, the use of affinity membrane to remove endotoxin in interferon preparations has high flow rate, good removal effect, and high recovery rate of active ingredients, which is better than column affinity chromatography.

具体实施方式Detailed ways

实施例1.Example 1.

制备以壳聚糖为配基的亲和膜:Preparation of affinity membrane with chitosan as ligand:

取50片直径为47毫米的纤维素膜,用大量水冲洗干净,置于一个200ml的烧杯中,加入40ml环氧氯丙烷、120ml 1M的NaOH 和0.19gNaBH4,混合均匀,室温下静置过夜,进行活化;次日取活化后的膜,用大量水洗至中性,沥干,然后加入150ml 1%壳聚糖(分子量35万)溶液(1%醋酸溶液配制),于60℃下过夜(16小时以上);次日取活化后的膜,用大量水洗至中性,沥干,然后加入100m10.25%硼氢化钠溶液(0.1M pH8.5磷酸缓冲液配制),室温下还原反应4小时,取还原后的膜,用大量水洗至中性,沥干并烘干。该亲和膜的含氮量为0.02%。Take 50 pieces of cellulose membranes with a diameter of 47 mm, rinse them with a large amount of water, place them in a 200ml beaker, add 40ml of epichlorohydrin, 120ml of 1M NaOH and 0.19g of NaBH , mix well, and let stand overnight at room temperature. Activation; take the activated membrane the next day, wash it with a large amount of water until neutral, drain, then add 150ml of 1% chitosan (molecular weight: 350,000) solution (prepared by 1% acetic acid solution), overnight at 60°C (16 Hours or more); the next day, take the activated membrane, wash it with a large amount of water until it is neutral, drain it, then add 100m10.25% sodium borohydride solution (prepared with 0.1M pH8.5 phosphate buffer), and perform a reduction reaction at room temperature for 4 hours , take the reduced film, wash it with a large amount of water until neutral, drain and dry. The nitrogen content of the affinity membrane is 0.02%.

以壳聚糖为配基的亲和膜的应用效果:Application effect of affinity membrane with chitosan as ligand:

将10张亲和膜制成相应的分离器,以9ml/min的流速过滤干扰素制剂200ml(干扰素效价91406IU/ml,溶液pH3.5),过滤后溶液的内毒素浓度从>80EU/ml降低到<10EU/ml,干扰素的活性(效价)回收率为90%。10 pieces of affinity membranes were made into corresponding separators, and 200ml of interferon preparations were filtered at a flow rate of 9ml/min (interferon titer 91406IU/ml, solution pH3.5), and the endotoxin concentration of the solution after filtration was from> 80EU/ ml is reduced to <10 EU/ml, and the recovery rate of interferon activity (titer) is 90%.

实施例2.Example 2.

制备以己二胺为配基的亲和膜:Preparation of affinity membrane with hexamethylenediamine as ligand:

取320片直径为47毫米的纤维素膜,用大量水冲洗干净,置于一个1000ml的烧杯中,加入650毫升氧化液中(含2%NaIO4),30℃烘箱过夜进行活化;次日,取活化后的膜,用大量水洗至中性,沥干,然后加入800ml1%己二胺(0.1M pH=7磷酸缓冲液配制),30℃烘箱过夜,进行交联反应;次日取交联后的膜,用大量水洗至中性,沥干,加入800ml 0.6%NaBH4(0.1MpH=9磷酸缓冲液配制)还原,30℃烘箱过夜。再用大量水洗至中性,沥干并烘干。该亲和膜的含氮量为2.2%。Get 320 pieces of cellulose films with a diameter of 47 mm, rinse them with a large amount of water, put them in a 1000ml beaker, add 650ml of oxidizing solution (containing 2% NaIO ), and activate them overnight in a 30°C oven; the next day, take The activated membrane was washed with a large amount of water until it was neutral, drained, and then 800ml of 1% hexamethylenediamine (prepared in 0.1M pH=7 phosphate buffer) was added to the membrane, and the cross-linking reaction was carried out in a 30°C oven overnight; the cross-linked membrane was taken the next day. The membrane was washed with a large amount of water until it was neutral, drained, added 800ml of 0.6% NaBH4 (prepared with 0.1MpH=9 phosphate buffer) for reduction, and baked at 30°C overnight. Wash again with plenty of water until neutral, drain and tumble dry. The nitrogen content of the affinity membrane is 2.2%.

以己二胺为配基的亲和膜的应用效果:Application effect of affinity membrane with hexamethylenediamine as ligand:

将10张亲和膜制成相应的分离器,以18ml/min的流速过滤干扰素制剂225ml(干扰素效价93976IU/ml,溶液pH4.2),过滤后溶液的内毒素浓度从>80EU/ml降低到<10EU/ml,干扰素的活性(效价)回收率为88%。10 affinity membranes were made into corresponding separators, and 225ml of interferon preparations were filtered at a flow rate of 18ml/min (interferon titer 93976IU/ml, solution pH4.2), and the endotoxin concentration of the solution after filtration was from> 80EU/ ml is reduced to <10EU/ml, and the recovery rate of interferon activity (titer) is 88%.

实施例3.Example 3.

制备以去氧胆酸钠为配基的亲和膜:Preparation of affinity membrane with sodium deoxycholate as ligand:

取50片直径为47毫米的纤维素膜,用大量水冲洗干净,置于一个200ml的烧杯中,加入40ml环氧氯丙烷、120ml 1M的NaOH和0.19克NaBH4,混合均匀,室温下静置过夜,进行活化;次日取活化后的膜,用大量水洗至中性,沥干,然后加入125ml 1%己二胺溶液(1%pH9磷酸缓冲液配制),于60℃下过夜(16小时以上);次日取膜,用大量水洗至中性,沥干,然后加入125ml 0.1M pH6.0的MES缓冲液,再加入2.5克去氧胆酸钠、1.25克EDC,混匀,室温下过夜。取交联后的膜,用大量水洗至中性,沥干并烘干。即得以去氧胆酸钠为配基的亲和膜。Take 50 pieces of cellulose membranes with a diameter of 47mm, wash them with a large amount of water, put them in a 200ml beaker, add 40ml of epichlorohydrin, 120ml of 1M NaOH and 0.19g of NaBH4, mix well, and let stand overnight at room temperature , to activate; the next day, take the activated membrane, wash it with a large amount of water until it is neutral, drain it, then add 125ml of 1% hexamethylenediamine solution (prepared with 1% pH9 phosphate buffer), overnight at 60°C (more than 16 hours) ); Take the membrane the next day, wash it with a large amount of water until it is neutral, drain it, then add 125ml of 0.1M MES buffer solution with pH 6.0, then add 2.5 grams of sodium deoxycholate and 1.25 grams of EDC, mix well, and leave overnight at room temperature . Take the cross-linked film, wash it with plenty of water until neutral, drain and dry. That is, the affinity membrane with sodium deoxycholate as ligand.

以去氧胆酸钠为配基的亲和膜的应用效果:Application effect of affinity membrane with sodium deoxycholate as ligand:

将10张亲和膜制成相应的分离器,以9ml/min的流速过滤干扰素制剂200ml(干扰素效价91406IU/ml,溶液pH6),过滤后溶液的内毒素浓度从>80EU/ml降低到<40EU/ml,干扰素的活性(效价)回收率为70%。10 affinity membranes were made into corresponding separators, and 200ml of interferon preparation was filtered at a flow rate of 9ml/min (interferon titer 91406IU/ml, solution pH6), and the endotoxin concentration of the filtered solution was reduced from > 80EU/ml To <40EU/ml, the recovery rate of interferon activity (titer) is 70%.

Claims (6)

1. from interferon formulation, remove endotoxic method for one kind, it is characterized in that: with chitosan, or sodium deoxycholate, or hexanediamine is as affinity ligand, be covalently bound on the porous medium, mode with dynamic filtration or Static Adsorption is operated, in order to remove the intracellular toxin in the recombinant interferon preparation; Wherein the amount of affinity ligand on porous medium is nitrogen content 0.02%~3%.
2. remove endotoxic method according to claim 1 is described from interferon formulation, it is characterized in that: described porous medium is to contain polyhydric polysaccharide media fibers element, dextran or agarose; Or contain polyhydric synthetic medium polyvinyl alcohol.
3. remove endotoxic method according to claim 1 is described from interferon formulation, it is characterized in that: the pH of interferon formulation is pH 3.5-6.0.
4. remove endotoxic method according to claim 1 is described from interferon formulation, it is characterized in that: be affinity ligand with the chitosan, the molecular weight ranges of chitosan is 10,000-1,000,000.
5. remove endotoxic method according to claim 1 is described from interferon formulation, it is characterized in that: the pattern of porous medium is flat sheet membrane, hollow-fibre membrane, bead or particle.
6. according to the described a kind of endotoxic method of removing from interferon formulation of claim 1, it is characterized in that: the flow velocity of described dynamic filtration interferon formulation is 1-20ml/min.
CNB031110363A 2003-02-21 2003-02-21 Method of eliminating endotoxin from interferon preparation Expired - Lifetime CN1324048C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CNB031110363A CN1324048C (en) 2003-02-21 2003-02-21 Method of eliminating endotoxin from interferon preparation

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CNB031110363A CN1324048C (en) 2003-02-21 2003-02-21 Method of eliminating endotoxin from interferon preparation

Publications (2)

Publication Number Publication Date
CN1523037A CN1523037A (en) 2004-08-25
CN1324048C true CN1324048C (en) 2007-07-04

Family

ID=34283355

Family Applications (1)

Application Number Title Priority Date Filing Date
CNB031110363A Expired - Lifetime CN1324048C (en) 2003-02-21 2003-02-21 Method of eliminating endotoxin from interferon preparation

Country Status (1)

Country Link
CN (1) CN1324048C (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101298468B (en) * 2007-04-30 2011-06-01 齐鲁制药有限公司 Method for removing endotoxin from protein medicine in one step
CN115739031B (en) * 2021-09-02 2024-05-28 中国科学院理化技术研究所 Application of chitosan or derivatives thereof in removal of endotoxin in gelatin

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1194179A (en) * 1997-03-26 1998-09-30 中国科学院大连化学物理研究所 Chemical modification of microporous nylon film and preparation of affinity film with histidine ligand

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1194179A (en) * 1997-03-26 1998-09-30 中国科学院大连化学物理研究所 Chemical modification of microporous nylon film and preparation of affinity film with histidine ligand

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
亲和膜用于去除医药及人血清白蛋白溶液中的内毒素 魏桂林等,色谱,第20卷第2期 2002 *
亲和膜用于去除医药及人血清白蛋白溶液中的内毒素 魏桂林等,色谱,第20卷第2期 2002;纤维素亲和层析膜用于去除医药制剂中的内毒素 魏桂林等,膜科学与技术,第20卷第4期 2000;纤维素亲和膜用于内毒素的去除 魏桂林等,分析化学研究报告,第29卷第10期 2001 *
纤维素亲和层析膜用于去除医药制剂中的内毒素 魏桂林等,膜科学与技术,第20卷第4期 2000 *
纤维素亲和膜用于内毒素的去除 魏桂林等,分析化学研究报告,第29卷第10期 2001 *

Also Published As

Publication number Publication date
CN1523037A (en) 2004-08-25

Similar Documents

Publication Publication Date Title
Zeng et al. Cross-linked macroporous chitosan anion-exchange membranes for protein separations
Shi et al. Functionalized anodic aluminum oxide (AAO) membranes for affinity protein separation
Ertürk et al. Cryogels-versatile tools in bioseparation
Ruckenstein et al. Macroporous chitin affinity membranes for lysozyme separation
Zeng et al. Membrane chromatography: preparation and applications to protein separation
EP2598223B1 (en) Chromatography media and method
US6860393B2 (en) Filter device and filter element comprising at least one negatively charged membrane
Suen et al. Exploiting immobilized metal affinity membranes for the isolation or purification of therapeutically relevant species
JP5396933B2 (en) Liquid chromatography packing and biopolymer separation and purification method
CN101835791A (en) protein purification method
MX2012001172A (en) Specific sorbent for binding proteins and peptides, and separation method using the same.
WO2009145722A1 (en) Separation method utilizing polyallylamine ligands
WO2000050160A9 (en) Negatively charged membrane
Avramescu et al. Particle-loaded hollow-fiber membrane adsorbers for lysozyme separation
Gondim et al. Dye ligand epoxide chitosan/alginate: a potential new stationary phase for human IgG purification
Pitiot et al. A potential set up based on histidine hollow fiber membranes for the extracorporeal removal of human antibodies
Vijayalakshmi Histidine ligand affinity chromatography
CN1324048C (en) Method of eliminating endotoxin from interferon preparation
WO2013187512A1 (en) Alkali-resistant ion exchange temperature-responsive adsorbent, and method for producing same
JP2021515698A (en) Composite material for bioseparation
Suen et al. Effects of spacer arms on cibacron blue 3GA immobilization and lysozyme adsorption using regenerated cellulose membrane discs
Machado et al. Evaluation of a chitosan membrane for removal of endotoxin from human IgG solutions
CN101060931B (en) Antibodies by chromatography
CN109078505A (en) A kind of composite chromatography filter membrane and its preparation method and application containing adsorbing medium
CN1277582C (en) Method for removing endotoxin from influenza vaccine preparations

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
C56 Change in the name or address of the patentee
CP03 Change of name, title or address

Address after: No. 3, building 6, Jingdong street, Beijing economic and Technological Development Zone

Patentee after: BEIJING KAWIN TECHNOLOGY SHARE-HOLDING Co.,Ltd.

Address before: No. 457, Zhongshan Road, Liaoning, Dalian

Patentee before: Dalian Institute of Chemical Physics, Chinese Academy of Sciences

C56 Change in the name or address of the patentee

Owner name: BEIJING KAIYIN SCIENCE CO., LTD.

Free format text: FORMER NAME: DALIAN INST OF CHEMICOPHYSICS, CHINESE ACADEMY OF SCIENCES

C41 Transfer of patent application or patent right or utility model
TR01 Transfer of patent right

Effective date of registration: 20090320

Address after: Building 3, building 6, Jingdong street, Beijing economic and Technological Development Zone, postcode: 100176

Co-patentee after: BEIJING KAWIN BIOTECHNOLOGY Co.,Ltd.

Patentee after: BEIJING KAWIN TECHNOLOGY SHARE-HOLDING Co.,Ltd.

Address before: Building 3, building 6, Jingdong street, Beijing economic and Technological Development Zone, postcode: 100176

Patentee before: BEIJING KAWIN TECHNOLOGY SHARE-HOLDING Co.,Ltd.

ASS Succession or assignment of patent right

Free format text: FORMER OWNER: BEIJING FURUI TIANCHENG BIOLOGICAL TECHNOLOGY CO., LTD.

Effective date: 20150211

C41 Transfer of patent application or patent right or utility model
C56 Change in the name or address of the patentee
CP03 Change of name, title or address

Address after: 3 building, No. 6, Jingdong street, Yizhuang economic and Technological Development Zone, Beijing 100176, China

Patentee after: BEIJING KAWIN TECHNOLOGY SHARE-HOLDING Co.,Ltd.

Patentee after: Beijing Furui Tiancheng Biotechnology Co.,Ltd.

Address before: 3 building, No. 6, Jingdong street, Beijing economic and Technological Development Zone, 100176

Patentee before: BEIJING KAWIN TECHNOLOGY SHARE-HOLDING Co.,Ltd.

Patentee before: BEIJING KAWIN BIOTECHNOLOGY Co.,Ltd.

TR01 Transfer of patent right

Effective date of registration: 20150211

Address after: 3 building, No. 6, Jingdong street, Yizhuang economic and Technological Development Zone, Beijing 100176, China

Patentee after: BEIJING KAWIN TECHNOLOGY SHARE-HOLDING Co.,Ltd.

Address before: 3 building, No. 6, Jingdong street, Yizhuang economic and Technological Development Zone, Beijing 100176, China

Patentee before: BEIJING KAWIN TECHNOLOGY SHARE-HOLDING Co.,Ltd.

Patentee before: Beijing Furui Tiancheng Biotechnology Co.,Ltd.

CX01 Expiry of patent term

Granted publication date: 20070704

CX01 Expiry of patent term