从人源大容量噬菌体抗体库中筛选抗生存素抗体

目的从大容量天然噬菌体抗体库中筛选抗生存素(survivin)的人源单链抗体(single chainfragment variable,scFv)并进行鉴定。方法以survivin为抗原,固化在抗原管上,通过吸附—洗脱—扩增过程,从大容量天然噬菌体抗体库中筛选特异性噬菌体抗体,并转染HB2151菌,经异丙基-β-D-硫代半乳糖苷诱导制备成可溶性抗体,采用酶联免疫吸附测定法对其抗原结合活性进行检测,指纹图谱分析法分析抗体基因的种类。结果经过4轮筛选,共获得21个与survivin结合的阳性克隆,其中10个特异性结合的克隆,指纹分析有8种不同的抗survivin的scFv基因,在这8种中有5种获得可溶性表达。结论利用噬菌体抗体库技术获得了特异性的人源抗生存素抗体,为其在今后肿瘤治疗中的应用奠定基础。

Cloning of human anti-survivin antibodies from phage antibody library

Objective To screen out the human anti-survivin scFv antibody from large phage library. Methods Panning of large phage library against survivin were conducted to select specific antibodies against survivin. The binding activities and specificities were tested by enzyme-linked immunosorbent assay method. Positive clones against survivin were analyzed by DNA fingerprint. Soluble scFvs were prepared by infecting E coli. HB2151 with the selected phage antibodies and using isopropyl-β-D-thiogalactopyranoside for induction. Results After 4 rounds of panning, 21 positive clones were obtained and 10 clones had specific binding ability to survivin. DNA fingerprinting of the 10 clones revealed 8 different stripes, of the 8 different stripes, 5 clones could express soluble survivin activity. Conclusion Human survivin antibodies were successfully obtained from large phage antibody library.