摘要
目的:制备全人源抗狂犬病病毒scFv-Fc融合抗体,并分析其结合活性。方法:构建全人源抗狂犬病病毒scFv-Fc融合抗体原核表达载体,优化表达并纯化全人源抗狂犬病病毒scFv-Fc融合抗体。以纯化的灭活狂犬病病毒包板,对纯化的抗体进行结合活性分析。结果:构建全人源抗狂犬病病毒scFv-Fc-pColdⅡ原核表达载体,经测序分析正确后,转入大肠杆菌BL21(DE3)进行表达,在加入浓度为0.5 mmol/L IPTG时,scFv-Fc融合抗体的表达较高,其分子量大小为57 000。纯化后的scFv-Fc融合抗体在1∶512的条件下仍可以较好地结合狂犬病病毒。结论:构建全人源抗狂犬病病毒scFv-Fc-pColdⅡ原核表达载体,表达并纯化了scFv-Fc融合抗体,为研发狂犬病暴露后预防治疗性抗体药物奠定了基础。
Objective：To prepare a human scFv-Fc fusion antibody against rabies virus and to analyze its binding activity.Methods：The human scFv-Fc prokaryotic expression vector was constructed.After optimizing prokaryotic expression system,the human scFv-Fc fusion antibody against rabies virus was expressed and purified.Purified humanized scFv-Fc antibody was confirmed its binding activity by binding to purified inactivated rabies virus.Results：After sequence analysis,the human scFv-Fc-pCold Ⅱprokaryotic expression vector was successfully established.Through transforming E.coli.BL21（DE3）,human scFv-Fc fusion antibody was expressed by optimized induction of IPTG at concentration of 0.5 mmol/L.The expression of scFv-Fc fusion antibody was increased and its molecular weight was 57 000.By purification,the human scFv-Fc fusion antibody was purified with excellent binding activity even in 512 times diluted concentration.Conclusion：We have successfully established a whole human anti-rabies virus scFv-Fc-pColdⅡ prokaryotic expression vector,and expressed and purified scFv-Fc fusion antibody.This fusion antibody can lay a foundation for developing novel antibody-targeted drugs of rabies prophylaxis after exposure.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2014年第6期741-744,749,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金资助(81273325)
江苏省科技支撑计划资助项目(BE2011842)
