摘要
目的：在大肠杆菌高效表达抗胃癌单抗３Ｈ１１的ｓｃＦｖ。方法：利用ＰＣＲ技术将３Ｈ１１ｓｃＦｖ基因从分泌型表达载体转移至高效表达载体，获得３Ｈ１１ｓｃＦｖ包含体的表达，经超声裂解、洗涤、变性后，尝试透析复性和凝胶过滤色谱柱上在位复性，结果：透析复性未能得到有活性的３Ｈ１１ｓｃＦｖ，而柱上在位复性效果良好，获得有功能性的３Ｈ１１ｓｃＦｖ。结论：成功进行了３Ｈ１１ｓｃＦｖ的柱上复性，不同ｓｃＦｖ在表达和复性中显示出明显差异，在基因工程抗体研制中值得注意。
Objective: To increase the yield of anti stomach cancer scFv, 3H11 in E coli . Methods: The 3H11 scFv gene was transferred from secretary expression vector to an insoluble expression vector with strong promoter. The expressed inclusion bodies were purified and solubilized. Renaturation of the solubilized scFv was conducted by both dialysis method and in situ refolding via gel filtration chromatography. Results: While the refolding by dialysis method failed the refolding employing gel filtration chromatography (Sephacryl S 200) succeeded and functional 3H11 scFv was obtained. Conclusion: In situ renaturation of scFv was a useful approach. The differences in scFv expression and refolding should draw attention in antibody engineering.
出处
《北京医科大学学报》
CSCD
1998年第6期506-508,共3页
Journal of Peking University(Health Sciences)
基金
国家"八六三"高技术项目
关键词
胃肿瘤
大肠杆菌
基因转移
单克隆抗体
Stomach neoplasms/genet Escherichia coli Gene transfer scFv inclusion bodies ☆ Antibodies, monoclonal
