三聚氰胺人源scFv抗体筛选与鉴定

Screening and Characterization of Human Phage Antibody to Melamine

目的筛选三聚氰胺人源单链可变区（scFv）抗体，为研究快速检测试剂盒奠定基础。方法采用噬菌体表面展示技术，分别以三聚氰胺、生物素化三聚氰胺作为包被抗原，从噬菌体抗体文库中经过3轮“吸附-洗脱-扩增”筛选过程，随机挑选阳性克隆，利用酶联免疫吸附法（ELIA）、交叉反应及竞争抑制实验，对比后获得与抗原结合活性较好的克隆提取质粒，亚克隆到pCANTAB5E载体，对其重组质粒进行分析。结果质粒酶切片段与目的相符。结论利用生物素化抗原筛选抗体创新了原有筛选方法，为下一步研究创造条件。

Objective Screening melamine in human single chain variable region （scFv） antibody,to lay the foundation for the study of rapid test kit. Methods Using phage display technology, melamine, biotinylated melamine as the coating antigen from a phage antibody library after three adsorption-elution-amplified the screening process, randomly selected positive clones,using enzyme-linked immunosorbent＇assay method （ELISA）, cross-reactive and competitive in hibition experiments, after comparing the antigen-binding activity better cloning plasmid and subcloned into pCANTAB5E vectors, recombinant plasmid analysis. Results Plasmid fragments and fit for purpose. Conclusion Used biotinylated antigen screening antibody innovation of the original screening methods to create conditions for further study.