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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。
OBJECTIVE: To screen and identify human scFv against soman transitional state analogues from high-capacity natural phage antibody library. METHODS: The phosphonate soman was used to hydrolyze the transition metal analogues penta-coordinate phosphonate and 3-hydroxy-1-p-nitrophenylphosphonic acid mono-pinacol ester. The process of de-amplification screened the specific antibody from the large-capacity natural phage antibody library and identified its antigen-binding activity. Results: After 4 rounds of screening, 14 positive clones against soman analogues were obtained respectively. According to DNA fingerprinting analysis, it was judged that the obtained clones contained four different scFv genes against soman analog respectively. Conclusion: The specific human anti-Soman analogue antibody was obtained by phage antibody library technology.