抗Ⅰ型鸭肝炎病毒VP1蛋白单克隆抗体表位的鉴定  

Identification of the epitope for a monoclonal antibody against the VP1 protein of duck hepatitis virus type Ⅰ

作　　者：吴晓颖[1,2] 葛铭 张云[2] 

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所动物细菌病研究室,哈尔滨150001

出　　处：《黑龙江畜牧兽医》2013年第2期15-18,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31072132);现代农业水禽产业技术体系岗位科学家专项(CARS-43-10);农业科学研究公益特殊基金项目(201003012)

摘　　要：为了筛选Ⅰ型鸭肝炎病毒(Duck viral hepatitis typeⅠ,DVH-Ⅰ)VP1蛋白的抗原表位,试验应用噬菌体展示随机12肽库试剂盒,以抗DHV-ⅠVP1蛋白单克隆抗体纯化IgG作为靶标,通过ELISA和竞争抑制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。结果表明:通过3轮生物淘洗后,目标噬菌体得到282倍富集;随机挑选15个克隆进行测序分析及ELISA和竞争抑制ELISA检测,其中有8个噬菌体克隆可以与抗DVH-ⅠVP1蛋白单克隆抗体高度特异性结合,8个克隆的共有氨基酸序列为GMMIP。说明共有序列GMMIP可能是单克隆抗体2D5所识别的VP1蛋白的线性表位。To screen the epitope of antigen against the VPI protein of duck hepatitis virus type ｜ （DHV - Ⅰ ）, this experiment used phage- display random 12 peptides library kit, and the immunoglobulin G （IgG） whieh purified with a monoclonal antibody against the VP1 protein of DHV -Ⅰ was used as a target, then the binding characteristics of the clones were identified and screened through the ELISA and competitive inhibition ELISA, and the DNA was extracted from the positive clone for sequencing analysis. The resuhs showed that 282 - fold enrichment was found in the target phage after throe rounds of biopanning. Fifteen phage clones were randomly selected for sequencing analysis, and other fifteen phage clones were randomly selected for the detections by ELISA and competitive inhibition ELISA, eight of which could combine with the monoclonal antibody against the VP1 protein of DHV - Ⅰ with high specific activity. The mutual amino acid sequence of the eight phage clones was GMMIP. The resuhs indicate that GMMIP may be a linear epitope of the VP1 protein recognized by monoclonal antibody 2D5.

关 键 词：Ⅰ型鸭肝炎病毒 抗VP1蛋白单克隆抗体 噬菌体展示 抗原表位 

分 类 号：S853.31[农业科学—临床兽医学]