大肠杆菌表达抗原结合片段的策略

背景：抗原结合片段在大肠杆菌中表达量低是制约其大规模生产和应用的主要因素,通过对宿主细胞、表达载体、表达条件和纯化条件等进行改造与优化,将有可能获得基因工程抗体的高效表达。目的：总结并介绍抗原结合片断抗体在大肠杆菌中的表达和纯化的最新策略。方法：由第一作者用计算机检索中国期刊全文数据库(CNKI：1990/2010)和Medline数据库(1990/2010),检索词分别为“大肠杆菌,抗原结合片段,表达,纯化,抗体”和“Escherichia coli,Fab,Expression,antibody,purification”,语言分别设定为中文和英文。从Fab抗体在大肠杆菌中表达方式及纯化2方面进行总结,对抗原结合片断抗体在大肠杆菌中最新的表达和纯化等方面进行介绍。结果与结论：共检索到125篇文章,按纳入和排除标准对文献进行筛选,共纳入30篇文章。结果表明抗原结合片段抗体在大肠杆菌中表达方式主要为细胞质表达和分泌表达,其纯化主要有金属离子螯和纯化和特异蛋白亲和纯化,抗原结合片断在大肠杆菌中表达的影响因素是多方面且相互关联的。目前研究对大幅提高抗原结合片断抗体在大肠杆菌中最新的表达量和最适纯化策略尚不一致。

Strategies for fragment antigen binding expression in Escherichia coli

BACKGROUND:Low-yields of antigen-binding fragment (Fab) restricts its full scale operation and application, which may be changed by remodeling or optimizing host cells, expression vectors, expression condition and purification conditions.  OBJECTIVE:To summarize and introduce new strategies about expression and purification of Fab antibodies in Escherichia coli (E. coli).METHODS:CNKI and Medline databases were searched by the first author using key words of “Escherichia coli, Fab, Expression, antibody, purification” in Chinese or English from year 1990 to 2010. The Fab antibody expression and purification in E. coli were summed up and new researches concerning Fab antibody expression and purification in E. coli were introduced.RESULTS AND CONCLUSION:Totally 125 papers were found and 30 papers were included. The results show that the Fab antibodies are mainly by cytoplasmic expression and secreted expression in E. coli. Its purification methods are mainly by metal ions and chelating affinity purification and specific proteins affinity purification. The influence factors concerning Fab expression in E. coli are correlated to each other. A substantial increase in current research on the latest Fab antibodies in E. coli expression and purification strategies are not consistent optimal, there is no best, only the most suitable.