Xuan Yan, Zheng Hong, Liang Chao
Department of Anesthesiology, The First Teaching Hospital of Xinjiang Medical University, Urumqi 830054, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2010 Oct 30;28(5):352-4.
The 7-mer phage random peptide libraries was screened with the specific anti-EgB polyclonal antibody. Sixty clones picked randomly were deduced by DNA sequencing and their amino acid sequences were compared with EgB. After 5 rounds of panning, the phage recovery rate increased from 4.15 x 10(-5) to 4.30 x 10(-2). 45 out of 60 selected clones were ELISA positive by anti-EgB polyclonal antibody with a positive rate of 75%, 45 positive clones were sequenced and the predicted amino acid sequences showed low homology to EgB. The reactivity of selected peptide and sera from patients of anaphylactic shock induced by Echinococcus granulosus was detected by ELISA. The result showed that the absorbance value (A410) of 45 positive clones detected by using serum samples from patients was twice higher than that by using 7 peptide library. The 45 positive clones exhibited specific binding to anti-EgB antibody. The peptides screening from 7-mer phage random peptide libraries may be the mimic epitopes of anaphylactic shock induced by E. granulosus.
用特异性抗EgB多克隆抗体筛选7肽噬菌体随机肽库。随机挑选60个克隆进行DNA测序推导，并将其氨基酸序列与EgB进行比较。经过5轮淘选，噬菌体回收率从4.15×10⁻⁵提高到4.30×10⁻²。60个筛选出的克隆中有45个被抗EgB多克隆抗体ELISA检测为阳性，阳性率为75%，对45个阳性克隆进行测序，预测的氨基酸序列与EgB的同源性较低。通过ELISA检测所选肽与细粒棘球绦虫致过敏性休克患者血清的反应性。结果显示，用患者血清样本检测45个阳性克隆的吸光度值（A410）比用7肽库检测的高出两倍。45个阳性克隆与抗EgB抗体表现出特异性结合。从7肽噬菌体随机肽库中筛选出的肽可能是细粒棘球绦虫致过敏性休克的模拟表位。
