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目的利用噬菌体肽库技术筛选华支睾吸虫童虫模拟抗原表位,观察影响筛选效率和噬菌体滴度的关键因素。方法收集华支睾吸虫感染童虫期大鼠血清,用正辛酸硫酸铵法提纯多克隆抗体,用提纯的多克隆抗体对噬菌体十二肽库进行3轮筛选。随机挑取噬菌体克隆,用ELISA法检测其特异性。结果经3轮免疫学筛选获得19个噬菌体克隆,ELISA测定有17个能被纯化的华支睾吸虫(童虫期)感染大鼠血清抗体识别。结论利用噬菌体肽库技术可以筛选出华支睾吸虫童虫模拟抗原表位,靶分子浓度以及噬菌体液的稀释度等因素是技术关键。
OBJECTIVE: To screen for the mimic epitopes of Clonorchis sinensis with phage peptide library technology and to observe the key factors affecting the screening efficiency and phage titer. Methods The sera of Clonorchis sinensis were collected and the polyclonal antibody was purified with ammonium octanoate sulfate. The phage displayed by the purified polyclonal antibody was selected by three rounds of screening. Phage clones were randomly picked and tested for specificity by ELISA. Results Nineteen phage clones were screened by 3 rounds of immunological screening and 17 were identified by ELISA in serum of clonorchis sinensis (sperm-infected) infected rats. Conclusion It is the key technology that the phage peptide library can be used to screen for the mimic epitopes, the concentration of target molecules and the dilution of phage fluids.