摘要
以抗旋毛虫Ts87蛋白单克隆抗体2A2作为靶标，对噬菌体展示随机十二肽库进行筛选。通过ELISA鉴定筛选克隆的结合特性，并对阳性克隆提取DNA进行测序分析。结果显示，经3轮生物淘洗后，目标噬菌体得到433倍富集。随机挑选20个克隆进行ELISA鉴定，其中有18个噬菌体克隆可以与单抗2A2特异性结合。测序分析发现，这18个克隆带有4种氨基酸序列，分别为：TPHPHIFYREAS、DWKAWTQMLDSY、WQIEYPTLHSLW和VSPYwHEYwsEL。经比对未发现这4种序列与Ts87蛋白序列有明显同源性。将这4株噬菌体克隆扩增后，经Western．blot鉴定均可被单抗2A2识别。结果表明本次筛选鉴定得到的4个噬菌体展示肽可能是旋毛虫Ts87蛋白的模拟抗原表位。
The monoclonal antibody 2A2 against Ts87 protein was used as a target in screening Ph. D. -12TM phage display peptide library and three rounds of bio-panning were conducted. The specific phage clones were analyzed for the binding ability with ELISA. Single strand DNA of positive clones were purified for DNA sequencing. After three rounds of bio-panning, the enriched phage was increased by 433 times. Twenty clones were selected randomly for ELISA, of which 18 clones showed specific ability of binding mAb 2A2. Four sequences of amino acid TPHPHIFYREAS, DWKAWTQMLDSY, WQIEYPTLHSLW and VSPYWHEYWSEL, were displayed in the 18 positive clones. No significant homologous sequences was found between the 4 sequences and Ts87 protein. The four amplified phage clones also could be recognized by mAb 2A2 with Western-blot. The results indicated that the peptides displayed by positive phage clones might be mimotopes of Ts87 protein.
出处
《寄生虫与医学昆虫学报》
CAS
2009年第4期193-197,共5页
Acta Parasitologica et Medica Entomologica Sinica
基金
基金项目：国家自然科学基金项目（No.30872201）
北京市自然科学基金项目（5092006）
国家科技重大专项项目（2008ZZX10004-011）：北京市教育委员会科技发展计划重点项目（kz200810025009）
北京市科委科技计划课题（Z080502036708012）
高等学校博士点科研基金项目（20060025003）
北京市教委学术创新团队和北京市高校拔尖人才项目（BAHED）
关键词
旋毛虫
单克隆抗体
生物淘洗
噬菌体展示肽库
抗原表位
Trichinella spirialis
Monoclonal antibody
Bio-panning
Phage display peptide library
Mimotope
