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.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
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)
关键词
SARS病毒
噬菌体展示
抗体库
SARS virus
phage display
combinatorial antibody library
