| 抗STa多克隆抗体的制备、鉴定和纯化 |
| |
| 引用本文： | 胡文丽,匡雪,简宗辉,王皓然,段锐锐,陈培富.抗STa多克隆抗体的制备、鉴定和纯化[J].中国畜牧兽医,2015,42(5):1082-1087. |
| |
| 作者姓名： | 胡文丽 匡雪 简宗辉 王皓然 段锐锐 陈培富 |
| |
| 作者单位： | 1. 云南农业大学动物科学技术学院, 昆明 650201;2. 云南省曲靖农业学校, 曲靖 655000 |
| |
| 基金项目： | 云南省高端科技人才引进计划项目(2009CI125) |
| |
| 摘 要： | 产肠毒素大肠杆菌(ETEC)可引起人及动物急性腹泻,其关键毒力因子为耐热肠毒素(ST)。由于ST是一种具有强烈毒性但缺乏免疫原性的小分子肽,如何在保持其免疫原性的同时减弱其毒性已成为制备ST类毒素疫苗的研究热点。本试验以利凡诺法提纯兔血清白蛋白(RSA),经0.2%戊二醛活化,再与人工合成的甲醇可溶性STa偶联,十二烷基硫酸钠—聚丙烯酰胺凝胶电泳(SDS-PAGE)结果显示获得分子质量约108.5 ku的完全抗原。用该偶联物免疫新西兰白兔4次,采集抗血清并用Protein A/G 琼脂糖小珠进行纯化,斑点酶联免疫吸附试验(dot-ELISA)和免疫印迹(Western blotting)结果显示抗血清效价均为1:400。试验结果表明抗STa多克隆抗体成功制备,这为筛选ST结构类似物做有益的铺垫。
|
| 关 键 词： | 耐热肠毒素(ST) 偶联物 多克隆抗体 |
| 收稿时间： | 2014-11-12 |
Preparation,Identification and Purification of Polyclonal Antibody against STa |
| |
| HU Wen-li,KUANG Xue,JIAN Zong-hui,WANG Hao-ran,DUAN Rui-rui,CHEN Pei-fu.Preparation,Identification and Purification of Polyclonal Antibody against STa[J].China Animal Husbandry & Veterinary Medicine,2015,42(5):1082-1087. |
| |
| Authors: | HU Wen-li KUANG Xue JIAN Zong-hui WANG Hao-ran DUAN Rui-rui CHEN Pei-fu |
| |
| Institution: | 1. College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China;2. Qujing Agricultural School in Yunnan Province, Qujing 655000, China |
| |
| Abstract: | Enterotoxic Escherichia coli (ETEC) can cause acute diarrhea in human and animals, and its key virulent factor is heat-stable enterotoxin (ST).ST is a peptide with small molecular weight and great toxicity but without immunogenicity, so how to keep its immunogenicity and reduce its toxicity has become a hot research topic for preparing ST toxoid vaccine.In this study, rabbit serum albumin (RSA) was purified by rivanol method and activated in 0.2% glutaraldehyde solution before RSA was conjugated to synthetic methanol-soluble STa, SDS-PAGE result showed that an approximate 108.5 ku complete antigen was obtained.New Zealand White rabbits were immunized with the conjugate four times, the serum was then collected and used to purify immunoglobulin by immunoprecipitation with Protein A/G Plus-Agarose.Dot-ELISA and Western blotting result showed that the titer of the antiserum both reached 1:400.These results indicated that anti-STa polyclonal antibody was successfully prepared, which provided the way for screening of ST structure analogues. |
| |
| Keywords: | heat-stable enterotoxin (ST) conjugate polyclonal antibody |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《中国畜牧兽医》浏览原始摘要信息 |
| 点击此处可从《中国畜牧兽医》下载免费的PDF全文 |
|
