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聚乙二醇伴随式离子交换层析分离重组乙肝病毒表面抗原 被引量:2
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作者 闭静秀 周卫斌 +4 位作者 李岩 黄永东 张焱 董爱华 苏志国 《生物工程学报》 CAS CSCD 北大核心 2005年第6期947-953,共7页
针对由中国仓鼠卵巢细胞(CHO)表达的多聚亚基蛋白HBsAg在离子交换层析过程中容易因亚基解离而导致蛋白解聚和丧失生物活性的难题,实验中选择聚乙二醇(PEG)作为保护剂伴随式(Polyethylene Glycol-Accompanied)离子交换层析分离纯化HBsAg... 针对由中国仓鼠卵巢细胞(CHO)表达的多聚亚基蛋白HBsAg在离子交换层析过程中容易因亚基解离而导致蛋白解聚和丧失生物活性的难题,实验中选择聚乙二醇(PEG)作为保护剂伴随式(Polyethylene Glycol-Accompanied)离子交换层析分离纯化HBsAg。实验表明,在流动相中加入1%PEG10000(WV)作为纯化伴侣,HBsAg的回收率由55%左右提高到80%以上,纯化倍数基本保持在12左右。对纯化产物进行SDS_PAGE分析表明,1%PEG10000的纯化伴侣伴随式离子交换层析能全部保留HBsAg的糖基化蛋白单体(27kD和30kD),高效液相色谱联用多角度激光散射(High Performance Size Exclusion Chromatography-Multiangle Laser Light Scattering,HPSEC-MALLS)进一步分析阐明了PEG能促使HBsAg颗粒尺寸分布更均一,结构更接近天然乙肝表面抗原。 展开更多
关键词 纯化伴侣 聚乙二醇 乙肝表面抗原 离子交换层析
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Single-step Purification of Molecular Chaperone GroEL by Expanded Bed Chromatography
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作者 佟晓冬 杨征 +1 位作者 董晓燕 孙彦 《Chinese Journal of Chemical Engineering》 SCIE EI CAS CSCD 2003年第4期460-463,共4页
Expanded bed adsorption (EBA) is an integrative downstream processing technique for the purification of biological substances directly from unclarified feedstock. In this study, molecular chaperone GroEL, an important... Expanded bed adsorption (EBA) is an integrative downstream processing technique for the purification of biological substances directly from unclarified feedstock. In this study, molecular chaperone GroEL, an important protein folding helper both in vivo and in vitro, was purified by the single-step EBA technique from the unclarified homogenate of recombinant E. coli cells. Compared with packed bed adsorption, the EBA technique provided a single-step approach to yield an electrophoretic purity of GroEL. After the homogenate loading and column washing in the expanded bed mode, the GroEL protein was recovered by stepwise salt-gradient elution in packed-bed or expanded-bed modes, respectively. The expanded-bed elution mode was found as efficient as the packed-bed mode in the purification of GroEL from cell disruptate. 展开更多
关键词 expanded bed adsorption molecular chaperone GROEL PURIFICATION
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