摘要
目的采用渗透压冲击预处理细菌以提高重组IGF-1融合蛋白纯化中金属螯合亲和色谱的效率。方法采用20%的蔗糖-EDTA缓冲液预处理经诱导表达的大肠杆菌,然后将表达产物经Ni-NTA-琼脂糖亲和色谱柱进行分离,观察蔗糖溶液处理对纯化效果的影响。结果经蔗糖-EDTA缓冲液预处理后Ni-NTA-琼脂糖色谱柱纯化蛋白的产量可以提高约2倍。结论渗透压冲击预处理可以有效提高金属螯合亲和色谱的效率。
Purpose This study aimed at evaluating the effect of osmotic shock on the efficiency of IMAC purification of recombinant IGF-1 in E.Coli.Methods After IPTG induction,the bacteria were treated by osmotic shock before expressed fusion protein was purified by Ni2＋ affinity chromatography.The purification efficiencies of IMAC were compared between the osmotic shock method and control method.Results Pre-treatment of bacteria by osmotic shock could increase the yield of recombinant IGF-1 by about 2 folds.Conclusion Our results demonstrate that osmotic shock can increase the efficiency of IMAC.
出处
《中国生化药物杂志》
CAS
CSCD
北大核心
2010年第4期235-237,240,共4页
Chinese Journal of Biochemical Pharmaceutics
基金
福建省教育厅科技项目(JA09115)
福建省自然科学基金重点项目(2008J0006)资助
关键词
金属螯合亲和色谱
渗透压冲击
重组蛋白
immobilized metal ion affinity chromatography
osmotic shock
recombinant protein
