摘要
目的为了获取人源性ＨＦＲＳ基因工程抗体。方法直接从人体ＰＢＬ构建人全套ＳｃＦｖ、ＶＨ噬菌体表面呈现文库，以ｐａｎｎｉｎｇ方法筛至四级文库，以ＥＬＩＳＡ方法鉴定各克隆抗汉坦病毒活性。结果获得了１４个ＳｃＦｖ阳性克隆和１１个ＶＨ阳性克隆。其中ＳｃＦｖ最高阳性克隆Ａ４１０值为０．４４，ＶＨ最高阳性克隆Ａ４１０值为０．５０。对最高阳性克隆进行了测序分析，证明连接和克隆正确，获得了抗ＨＴＶ人源性ＳｃＦｖ和ＶＨ抗体的基因片段。克隆再转化于ＨＢ２１５１菌株，成功地进行了分泌型抗体片段的表达。结论采用噬菌体抗体库技术直接从人体克隆人源性汉坦病毒噬菌体抗体可行。
Objective To obtain the genetic engineering antibody against HFRS virus.Methods Repertoires of human ScFv and V H displayed on pHEN1 have been generated directly from human PBL. The two repertoires were panned into grade 4 repertoires. The activity of each clone to bind HTV was examined by using ELISA. Results 14 positive ScFv clones and 11 positive V H clones have been found . The highest value of A 410 in positive ScFv clones was 0 44, and in positive V H clones was 0 50 The two clones were sequenced, and the sequencing results demonstrated that the genes of ScFv and V H were the human antibody genes, and the gene ligating, cloning and screening were correct and successful. The secretory expression of ScFv and V H were also carried out. Conclusions The experimental results suggest that the skills used in the paper are good to obtain the genetic engineering antibedy against HFRS.And the antibody could be used for prevention and treatment of HFRS.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
1999年第1期38-41,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金
关键词
噬菌体
抗体
汉坦病毒
单链DNA
免疫球蛋白
Phage antibody Hantavirus Single chain Fv Variable region of antibody
