摘要
目的 构建抗 HFRS病毒 m Ab 1 A8鼠 /人嵌合 Fab噬菌体抗体表达载体 ,并进行表达 .方法 将抗 HFRS病毒m Ab1 A8的 VH 基因和人重链 CH1基因连接 ,VL 基因和人Cκ 基因连接 ,分别构建了鼠 /人嵌合重链 Fd基因和轻链基因 ,将它们克隆入 p Comb3,构建了鼠 /人嵌合 Fab噬菌体抗体表达载体 .用辅噬菌体 VCSM1 3超感染 ,EL ISA间接夹心法检测抗体活性 .结果 经多种限制性内切酶酶切鉴定载体构建正确 ,ELISA间接夹心法检测超感染上清为阳性 .结论 成功构建了鼠 /人嵌合 Fab噬菌体抗体表达载体 ,并在噬菌体表面表达出可结合 HFRS病毒抗原的鼠 /人嵌合
AIM To construct and to express the mouse/human chimeric Fab antibodies on phage. METHODS Mouse/human chimeric heavy chain Fd (Fd fragment) and light chain genes were generated by ligation of V H gene of mouse mAb 1A8 against HFRS virus with human heavy chain CH1 gene and V L gene with human C κ gene, respectively. The chimeric Fab expression vector was constructed by cloning the chimeric heavy chain Fd and light chain genes into pComb3. Fab antibodies were expressed on phage by superinfection with helper phage VCSM13, and the binding activity of antibodies was detected by indirect sandwich ELISA. RESULTS Mouse/human chimeric Fab expression vector was constructed successfully, which was identified by restriction enzymes digestion. The supernatant of superinfection could be detected that having activity of binding HFRS virus antigen by the indirect sandwich ELISA. CONCLUSION Mouse/human chimeric Fab antibody expression vector was constructed and expressed on the surface of phage successfully. The Fab antibodies had binding activity against HFRS virus antigen.
出处
《第四军医大学学报》
北大核心
2002年第4期324-326,共3页
Journal of the Fourth Military Medical University
基金
国家自然科学基金资助项目 ( 3970 0 134
39570 6 6 1)
关键词
肾综合征出血热病毒
嵌合抗体
单克隆
噬菌体抗体
hemorrhagic fever with renal syndrome virus
chimeric antibodies
antibodies, monoclonal
