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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。
OBJECTIVE: To construct a phage display library of human antibody against Fab fragment of SARS virus and screen for anti-SARS virus-specific phage antibodies. Methods: The human Fab fragment antibody gene was amplified by PCR and ligated into vector pComb3 to construct the phage antibody library. The antibody library was screened by immobilized SARS antigen and the specificity of phage antibody binding to SARS virus was detected by ELISA. Results: 13 Ig gene fragments were amplified by PCR. The phage antibody library constructed by electrotransformation has a capacity of 1. 3 × 106, and the recombination rate of the Fab gene is 60%. The purified SARS antigen was screened for 3 rounds, and the phage antibody against SARS virus was specifically enriched, and 10 clones with good binding activity and strong specificity were screened out by ELISA. Conclusion: The combinatorial antibody library of human anti-SARS virus has been successfully constructed, and the specific anti-SARS virus-specific antibody was obtained.