摘要
目的获得泰泽氏病原体抗原表位相关肽,用于实验动物血清中该病原体感染相关抗体的检测。方法选用泰泽氏病原体的四种单克隆抗体(M2、M3、M4、M5)作为配基,从噬菌体表面展示的随机7肽文库中筛选单抗识别的抗原表位,获得特异性噬菌体克隆;并采用ELISA、Western blot方法对其进行分析鉴定,获得阳性噬菌体克隆。结果获得阳性噬菌体克隆5个,其展示的融合蛋白能被泰泽氏病原体的免疫血清识别,ELISA检测A值的P/N为8.0～17.1;Western blot分析显示单一特异性条带,相对分子质量约为38×103。结论本研究获得的5个阳性克隆所表达的融合蛋白,为泰泽氏病原体抗原表位相关肽,可作为该病原体隐性感染血清学检测的候选抗原。
Objective To obtain antigenic epitope-associated peptides of Tyzzer＇s organism, which can be used to detect Tyzzer＇ s infection-related antibodies in the sera of laboratory animals. Methods Four monoclonal antibodies （ M2, M3 M4 and M5） of Tyzzer＇s organism were used as ligands to screen antigenic epitope-associated peptides of Tyzzer＇s organism by biopanning a phage-display random peptide library （RPL） of 7 ammino-acid residues. The specific-binding phage clones were obtained through three rounds of biopanning. The reactivity of positive phage clones with immune-serum of Tyzzer＇ s organism was identified by ELISA and Western blot assay. Results Five positive phage clones, which displayed fusion protein that could be specifically recognized by immunized-serum of Tyzzer＇ s organism, were obtained from RPL. The A value ratio of positive to negative varied from 8.0 to 17.1 by ELISA. A specific protein band located at relative molecular weight 38×10^3 or so was demonstrated in Western blot assay. Conclusion The fusion protein displayed on positive clones was confirmed as antigenic epitope-associated peptides of Tyzzer＇ s Organism, which could be used as a candidate of serodiagnostic antigen for detection of Tyzzer＇ s subclinical infection
出处
《中国实验动物学报》
CAS
CSCD
2005年第4期231-234,共4页
Acta Laboratorium Animalis Scientia Sinica
关键词
肽库
泰泽氏病原体
抗原
表位
Phage random peptide library （RPL）
Tyzzer＇ s Organism
Antigen
Epitope
