摘要
目的：构建人源性 Ｆａｂ 噬菌体抗体基因库并筛选可表达与肾综合征出血热（ Ｈ Ｆ Ｒ Ｓ）病毒抗原特异结合之 Ｆａｂ 抗体的重组克隆． 方法：利用逆转录、聚合酶链反应（ Ｐ Ｃ Ｒ）等技术从 Ｈ Ｆ Ｒ Ｓ恢复期患者外周血淋巴细胞（ Ｐ Ｂ Ｌ）中扩增出人 Ｉｇ Ｇ抗体重链 Ｆｄ 基因及κ轻链基因，将之克隆入噬菌体载体ｐ Ｃｏｍ ｂ３，构建人源性 Ｆａｂ 噬菌体抗体基因库． 并利用噬菌体表面呈现技术，对此抗体库进行淘筛、富集． 结果：①成功地构建了人源性 Ｆａｂ 抗体基因库，库容量为 ３×１０６ ． ②从噬菌体抗体库中淘筛得到了能表达人源性抗 Ｈ Ｆ Ｒ Ｓ病毒 Ｆａｂ 抗体的重组克隆． 结论：利用噬菌体抗体库技术，可以获得能表达人源性抗 Ｈ Ｆ Ｒ Ｓ病毒抗体的重组克隆．
AIM: To construct a human Fab phage antibody library and to obtain some recombinant clones which can express Fab antibodies against HFRS virus. METHODS: The total cell RNA extracted from peripheral lymphocytes of HFRS patient was transcribed reversely into cDNA. By amplifying the heavy chain Fd genes and light chain genes of human antibodies from cDNA using semi nested PCR and cloning them into pComb3 vector, a human library of Fab antibodies was constructed, and then it was panned by phage display technology. RESULTS:①A human library of Fab antibodies was constructed, its size is 3×10 6 members. ②Six clones which can express Fab antibodies against HFRS virus were obtained by panning. CONCLUSION: Some recombinant clones which express Fab antibodies against HFRS virus can be obtained by phage display technology.
出处
《第四军医大学学报》
1999年第7期575-577,共3页
Journal of the Fourth Military Medical University
基金
国家自然科学基金
