人源抗SARS病毒噬菌体抗体库的构建及筛选  

Construction and screening of human phage antibody library against SARS virus

作　　者：康晓平[1] 熊伟[1] 常国辉[1] 祝庆余[1] 

机构地区：[1]军事医学科学院微生物流行病研究所全军微生物检测研究中心,北京100071

出　　处：《细胞与分子免疫学杂志》2005年第1期46-49,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：军事医学科学院科技创新研究启动基金资助(No. 0208075)

摘　　要：　目的: 构建人源抗SARS病毒的Fab片段抗体基因的噬菌体表面呈现文库, 筛选抗SARS病毒特异性的噬菌体抗体。方法: 用PCR扩增人Fab片段抗体基因, 连入载体pComb3内, 构建噬菌体抗体库。以固相化的SARS抗原淘筛抗体库, 并用ELISA检测噬菌体抗体结合SARS病毒的特异性。结果: 用PCR共扩增出 13条Ig基因片段。电转化构建的噬菌体抗体库的库容为 1. 3×106, Fab基因的重组率为60%。以纯化的SARS抗原淘筛 3轮, 特异性富集了抗SARS病毒的噬菌体抗体, 并用ELISA法从中筛选出了 10个结合活性好、特异性强的克隆。结论: 成功地构建了人源抗SARS病毒的组合抗体文库, 从中获得人源抗SARS病毒的特异性抗体。AIM: To construct human phage antibody library against SARS virus. METHODS: Human Fab genes were amplified by RT-PCR from lymphocytes of a convalescent SARS patient and cloned into vector pComb3 to construct phage antibody library. Antibodies against SARS virus were screened by biopanning with immobilized virus antigen. The binding specificity of phage antibody to SARS virus was detected by ELISA. RESULTS: 13 Ig genes were obtained by PCR. A human phage antibody library consisting of 1.3×106 clones was constructed by electrotransformation and 60 percent of the clones contained Fab genes. After three rounds of panning with SARS virus antigen, phage antibodies against SARS virus were specifically enriched. ELISA detection verified 10 positive phage antibodies that were highly specific for SARS virus. CONCLUSION: Human phage antibody library against SARS virus has been constructed successfully, from which 10 anti-SARS virus antibodies were obtained.

关 键 词：SARS病毒 噬菌体展示 抗体库 

分 类 号：Q782[生物学—分子生物学]