摘要
目的:采用膜亲和色谱这一新型纯化分离手段,结合膜分离过程的高速度和亲和吸附的高灵敏度进行水系溶液中内毒素的吸附去除工作,以求建立一种方便实用的内毒素去除方法。方法:选取优质滤纸为原料,交联后制成纤维素膜,分别用静态和动态两种方法制备出含有组氨酸和粘多菌素的亲和膜。结果:以组氨酸为配基的亲和膜,其吸附效果可使滤液中的内毒素水平低至0.5EU·ml-1以下,可满足一般医药卫生方面的要求;以粘多菌素B为配基的亲和膜,其吸附能力好于组氨酸亲和膜,对自来水和药物中天然内毒素的去除率在95%以上。结论:亲和膜堆对内毒素的去除效果与文献报道的柱亲和色谱法相当。但在成本、效率、处理量及使用寿命方面更具有优越性和实用性。
Abstract
OBJECTIVE:Membrane affinity chromatography was adopted for the removal of endotoxin from aquoeous solutions This technique which combine the processing capacity of membrane separation and selectivity of affinity adsorption,proved to be efficient and convenient for the removal of endotoxin from solutions METHODS:Macropore cellulose membrane was prepared and then derived chemically before histidine or polymyxin B were immobilized as affinity ligands Affinity membrane cartridges were also prepared using two sealing methods according to different ligands adopted Onto a tape sealed cartridge,dynamic immobilization of affinity ligand was also performed,which was much favourable for the application of biomacromolecular ligand RESULTS:The affinity membrane cartridges prepared were used for the removal of endotoxin from aqueous solutions By a cartridge with 40 sheets of affinity membrane,the endotoxin content in solution could be reduced to a minimum of 0.4 EU·ml-1 CONCLUSION:Compared with the classic column method,the membrane medium was more cheap,durable and easy to scaled up It is hopeful that membrane affinity cartridge will find its big application in the purification of pharmaceutics and biomolecular products.
关键词
亲和膜 /
内毒素 /
组氨酸 /
粘多菌素B
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Key words
affinity membrane /
endotoxin removal /
histidine /
polymyxin B
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郭为;商振华;于亿年;周康;周良模.
大孔纤维素亲和膜在生化制剂纯化中的应用[J]. 中国药学杂志, 1998, 33(12): 731-734
Guo Wei.
Preparation of macropore cellulose membrane and its application in the endotoxin removal by affinity adsorption [J]. Chinese Pharmaceutical Journal, 1998, 33(12): 731-734
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参考文献
1 Hou K C,Zaniewski R.Depyrogenation by Endotoxin Re-moval with Positively Charged Depth Filter Cartridge.J Parenter Sci Technol,1990,44(4):204.
2 Minobe S,Watanabe T,Sato T,et al.Preparation of Adsor-bents for Pyrogen Adsorption.J Chromatogr,1982,248:401.
3 Issekutz A C.Removal of Gram-Negative Endotoxin from Solurtions by Affinity Chromatography.J lmmunol Methods,1983,6l:275.
4 Guo W,Shang ZH,Yu YN,et al.Membrane Affinity Chro-matography of Alikaline Phosphatase.J Chromatogr,1994,685:344.
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脚注
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