从噬菌体随机肽库中淘选模拟NFLX(Norfloxacin)表位的噬菌体粒子并进行性质鉴定。以抗NFLX单克隆抗体为靶分子,对噬菌体环七肽库和十二肽库进行生物亲和淘选,以ELISA鉴定阳性克隆,进行DNA测序,采用生物信息学方法对淘选得到的模拟表位进行分析。分别得到了9个(环七肽)和8个(十二肽)能与靶分子特异性结合且能被NFLX标准品阻断的阳性克隆,其特有序列为(环七肽):X-X-X-X-X-脯氨酸-苯丙氨酸(X为任意氨基酸),以阻断效果最好的抗原模拟表位H3建立了竞争ELISA标准曲线,IC50为:(48.07±6)ng·mL^-1,检测线性范围为10-200ng·mL^-1。噬菌体展示技术可淘选得到NFLX模拟表位,淘选到的噬菌体粒子可作为NFLX的替代品建立免疫学检测方法。
To screen phage particles capable of mimicking norfloxacin (NFLX) and identify the properties. The anti-NFLX monoclonal antibody was used as a target for biopanning from c7c phage display peptide library and phage 12-mer peptide library. The positive clones were identified by ELISA and sequenced, then analyse the obtained mimotopes by means of the bioinformatics. 9 positive clones(c7c phage display peptide library)and 8 positive clones(phage 12-mer peptide library)can bind with anti-NFLX monoclonal antibody and be blocked by NFLX,they have common feature(c7c phage display peptide). X-X-X-X-X-prolin-phenylalanine (X can be any amino acid), H3 was used to establish a competitive ELISA immunoassay,the linear range of the inhibition curves was between 10 ng . mL^-1 and 200 ng. mL^-1 ,IC50 was 48.07±6 ng. mL^-1. The phage display technique can be successfully applied to panning the mimotope of Norfloxacin. The aquired phages may be used as the surrogate of the toxin to establish immunoassay.