摘要
将抗ＨＢｓＡｇ 抗体的轻链（Ｌ）和重链Ｆｄ 段基因，分别克隆于ｐＥＴ２０ｂ 质粒中并分别转化到大肠肝菌ＢＬ２１（ＤＥ３），ＬＰＴＧ诱导后，ＳＤＳ－ＰＡＧＥ分析发现在２７ＫＤ（Ｌ）和２５ＫＤ（Ｆｄ）处有外源蛋白表达，表达蛋白含量分别为５３％ 和４８％ 。Ｌ链和Ｆｄ 包涵体蛋白经盐酸胍变性后，等量混合于折叠液中，Ｌ和Ｆｄ 可复性形成了约５０ＫＤ的蛋白。ＥＬＩＳＡ结果表明，复性蛋白具有与ＨＢｓＡｇ 结合的能力。抗ＨＢｓＡｇ 抗体Ｆａｂ 段在大肠杆菌中的表达与复性的成功，表明包涵体表达基因工程抗体在技术是可行的。
The genes of Fd fragment and L chain of human antibody against hepatitis B virus surface antigen(HBsAg) were cloned into plasmid pET 20b. The resultant plasmids of pETFd and pETL were transformed into E.coli BL21(DE3) respectively, resulting in production of a 25KD and a 27KD protein by the induction with IPTG. The expression level of Fd and L protein was 48% and 53%. After Fd fragment and L chain inclusion bo dies were solubilized and combined in equal molar ratio in the refolding solution. 50 KD protein was observed by SDS PAGE and its antigenicity was confirmed by western blot. Furthermore, the binding activity to HBsAg was revealed by an ELISA. These results indicated that the inclusion strategy of producing Fab fragment was available in technology.
出处
《免疫学杂志》
CAS
CSCD
北大核心
1999年第4期229-231,共3页
Immunological Journal
基金
国家"863"计划资助
