摘要
目的:将大肠杆菌表达的抗乙肝表面抗原(HBs)人Fab转换为真核表达的全长人lgG1.方法:用重叠PCR法将人工合成的人V_k前导序列拼接到抗HBs的VH和V_k上,构建人IgGl真核表达载体,转染真核细胞,通过ELISA、RT-PCR、免疫印迹检测抗HBs人IgG的表达.结果:用轻链和重链同时表达的单一载体在CHO细胞中获得了抗HBs人lgG表达.结论:通过噬菌体抗体库所获Fab段可通过拼接人V_k前导序列在真核获得功能性表达.
Objective: Conversion of anti-HBs human Fab derived from phage antibody library to eukaryoticexpressed whole human IgG molecules. Methods: A synthesized human VK leader sequence was spliced to anti-HBs VH and VK by overlap are. Eukaryotic expression vectors were constructed and transfected into SP2/0 orCHO cells. The expression of anti-HBs human IgG was tested by HLISA, RT-PCR and Western blot. ResultAnti-HBs human IgG was expressed by transfecting CHO cells with a single vector tha contained both lightchain and heavy chain genes. Conclusion:Fabs obtained for phage antibody library could be expressed in eu-karyotic cells using a human VK leader sequence.
出处
《海军总医院学报》
1999年第1期1-5,共5页
Journal of Naval General Hospital of PLA
基金
国家863资助项目(Z-18-01-02-03)
