人源抗梭曼过渡态类似物(P6)的抗体筛选

Cloning of human scFv against soman transitionstate analogues(P6) from a large phage antibody library

目的:从大容量天然噬菌体抗体库中筛选抗梭曼过渡态类似物的人源单链抗体(scFv)并进行鉴定。方法:以膦酸梭曼水解过渡态类似物五配位-羟基膦酸酯,3-羟基-1-对硝基苯基甲膦酸单频哪基醇酯,为半抗原,通过吸附-洗脱-扩增过程从大容量天然噬菌体抗体库中筛选特异性抗体,对其抗原结合活性进行鉴定。结果:经过4轮的筛选,分别获得14个抗梭曼类似物的阳性克隆。经DNA指纹分析,判断所获克隆分别包含4个不同的抗梭曼类似物的scFv基因。结论:利用噬菌体抗体库技术获得了特异性的人源抗梭曼类似物抗体。

AIM：To clone human anti-soman transitionstate analogues antibodies from a large single-chain phage antibody library.METHODS：An organophosphorus hapten P6 [O1-methyl-O2-（1,2,2-trimethylpropyl）-2-hydroxy-5-nitrophenyl methylphosphonic acid] was synthesized.Its chemical conjugate with bovine serum albumin（BSA） was used as antigen（P6-BSA） to screen antibodies against soman.Panning of a large single-chain phage antibody library was conducted to select specific antibodies against soman.The antigen binding characteristics were analyzed by ELISA.RESULTS：After 4 rounds of panning,14 clones had specific binding ability to P6.DNA fingerprinting showed that diverse specific human scFvs against P6 was obtained from the library by biopanning.CONCLUSION：Human anti-soman transitionstate analogues scFvs have been cloned from large phage antibody library.