不同色谱介质用于鼠腹水中低等电点单克隆抗体的纯化  

Purification of Low PI McAb in Murine Ascites with Different Chromatography Media

作　　者：蒋先敏[1] 孔健[1] 李宇静[1] 赵莉[1] 曹竹安[2] 

机构地区：[1]卫生部北京生物制品研究所,北京100024 [2]清华大学化学工程系,北京100084

出　　处：《中国生物制品学杂志》1998年第2期80-83,共4页Chinese Journal of Biologicals

基　　金：国家九五攻关项目!96-C02-03-05

摘　　要：从一低等电点单克隆抗体（C50，CEA单抗）鼠腹水中纯化单抗，对10种色谱介质在其离子强度、pH、流速等最佳的条件下的纯化效果进行了对比，结果表明腹水先用40％饱和硫酸铵处理后，经一次色谱柱纯化，纯度可达83．6％～96％，回收率为50％～94％。在各种阴离子交换介质中，QSepharoseFF的效果最好，弱阴离子交换介质WhatmanDE52的效果较差。尽管两种强阳离子交换介质的效果较好，但劣于QSepharoseFF及亲会色谱介质HiTrapProteinG。Poros50A纯化IgG1类单抗时需要较高的盐浓度，不能作为最佳候选介质。SephacrylS-200HR及PhenylSepharose6FF纯化产物回收率及纯度均较低，不适于该单抗的纯化。The purification effects of low PI McAb in murine ascites with 10 kinds ofchromatography media under suitable conditions(ionic strength,pH,flow rate and so on)were compared.The result showed that the purity of McAb in ascites reached 83.6%～96%after being treated with 40% saturated(NH4)2SO4 and purified by column chromatography,and the recovery rate of it reached 50%～94%.Of the anion exchange media,Q Sepharose FFshowed good purification effect,however,the purification effect of Whatrnan DE52 was bad.Though 2 kinds of strong cation exchange media showed good purification effects,the effectsof them were worse than those of Q Sepharose FF and HiTrap Protein G.The purification ofMcAb to IgG1 by Poros 50A needed a high salinity,so the media could not be used as the bestcandidate.The recovery rates and purities of McAbs purified by Sephacryl S-200HR andPhenyl Sepharose 6FF were low,so the 2 kinds of media were not suitab1e for the purificationof McAb in ascites.

关 键 词：单克隆抗体 色谱介质 纯化 

分 类 号：R392－33[医药卫生—免疫学]