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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: To transform E. coli-derived human anti-hepatitis B surface antigen (HBs) Fab into full-length human lgG1 with eukaryotic expression.Methods: The synthetic human V_k leader was spliced to VH and V_k of anti-HBs by overlapping PCR The eukaryotic cells were transfected with eukaryotic cells and the anti-HBs human IgG expression was detected by ELISA, RT-PCR and Western blotting.Results: A single vector with both light and heavy chain expression was obtained in CHO cells Anti-HBs lgG expression was observed.Conclusion: The Fab fragment obtained from the phage antibody library can be obtained in eukaryotic expression by splicing human V_k leader sequence.