摘要
目的：在大肠杆菌中高效表达抗乙肝表面抗原的单链抗体。方法：通过ＰＣＲ将单链抗体基因重组到原核细胞表达载体ｐＴ７中，构建单链抗体高效表达载体ｐＴ７ＳＣ。将ｐＴ７ＳＣ质粒转化大肠杆菌ＢＬ２１（ＤＥ３），ＩＰＴＧ诱导进行表达。结果：获得了ＳｃＦｖ的高效表达，表达产物占菌体总蛋白的２８％以上。竞争抑制性ＥＬＩＳＡ检测诱导后的细菌培养上清，证明所表达的ＳｃＦｖ具有抗体活性。进一步将ｃＭｙｃ十肽连接在ＳｃＦｖ的羧基端，使所表达的ＳｃＦｖ易于检测。结论：在大肠杆菌高效表达抗乙肝ＳｃＦｖ。
Objective:To obtain high level expression of anti HBsAg ScFv in E.coli.Methods:An anti HBsAg ScFv expression vector pT7SC was constructed by inserting the ScFv gene obtained by PCR amplification from vector pHBSCS constructed previously into a T7 promoter containing vector. pT7SC was transformed into E.coli BL21(DE3) cells and expression was induced by IPTG.Results:The ScFv product constituted 28% of the total bacterial protein.HBsAg binding activity was detected in the supernatant of the induced bacteria by competitive ELISA.In order to facilitate the detection of ScFv,the vector was further modified attaching sequences encoding c Myc derived decapeptide to the 3′ end of ScFv gene.ScFv protein thus produced could be easily detected by ELISA using McAb 9E10.Conclusion:Success in high level expression of anti HBsAg ScFv would facilitate the use of small antibody molecules.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
1999年第3期122-123,共2页
Chinese Journal of Immunology
基金
国家科委"863"基金
