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目的构建VH 基因随机CDR3ScFv噬菌体抗体库 ,并筛选抗HFRS病毒NP抗原的噬菌体抗体。方法采用随机引物PCR法扩增抗HFRS病毒mAb1A8的VH 基因 ,并与1A8VL 基因共同克隆入噬菌粒表达载体 pHEN1后 ,构建VH 基因随机CDR3ScFv基因库。继用辅噬菌体VCSM13超感染后得到噬菌体抗体库 ,然后用HFRS病毒抗原进行筛选。随机挑取筛选的菌落超感染 ,用夹心ELISA检测超感染上清。结果获得库容量约为107 噬菌体抗体库。夹心ELISA的结果表明 ,筛选出的噬菌体抗体可与HFRS病毒NP抗原特异性结合。结论成功地构建了库容量较高的ScFv噬菌体抗体库 ,并获得可与HFRS病毒NP抗原结合的噬菌体抗体。
Objective To construct VH CDR3 ScFv phage antibody library and screen for anti-HFRS virus NP antigen phage antibody. Methods VH gene of anti-HFRS virus mAb1A8 was amplified by random primer PCR and cloned into phagemid expression vector pHEN1 with 1A8VL gene to construct random CDR3ScFv gene library of VH gene. Following phagemid VCSM13 super-infection obtained phage antibody library, and then screening with HFRS virus antigen. Colonies were randomly picked and screened for superinfected supernatants by sandwich ELISA. As a result, a pool of about 107 phage antibody libraries was obtained. Sandwich ELISA results showed that the screened phage antibodies could specifically bind to the HFRS virus NP antigen. Conclusion The library of ScFv phage antibody with high library capacity was successfully constructed and the phage antibody that could bind to the NP antigen of HFRS virus was obtained.