| 抗恩诺沙星噬菌体单链抗体库的构建及鉴定 |
| |
| 引用本文： | 温凯,沈建忠,孟辉,吴聪明,王战辉,张素霞.抗恩诺沙星噬菌体单链抗体库的构建及鉴定[J].中国农业大学学报,2011,16(2):118-124. |
| |
| 作者姓名： | 温凯 沈建忠 孟辉 吴聪明 王战辉 张素霞 |
| |
| 作者单位： | 中国农业大学,动物医学,北京,100193 |
| |
| 基金项目： | 国家自然科学基金重点项目(30830082); 国家“863”计划项目(2008AA10Z423) |
| |
| 摘 要： | 本研究以兽药恩诺沙星为对象,构建噬菌体抗体库,为低成本、快速制备目的抗体提供新的途径。以恩诺沙星-鸡卵清蛋白(ENR-OVA)为免疫原对Balb/c小鼠进行免疫,取其脾细胞提取总RNA,并分别扩增全套抗体轻、重链基因,通过重叠延伸PCR技术,以编码柔性多肽(Gly3Ser)4的基因为接头,将轻重链基因组装为完整的scFv基因,将之克隆入pCANTAB5E载体,转化大肠杆菌XL1-Blue,以辅助噬菌体M13KO7对其进行超感染,构建噬菌体抗体库并进行富集、筛选和鉴定;构建了库容量约为2.2×106的抗恩诺沙星噬菌体单链抗体库,并筛选出26株阳性克隆,为表达单链抗体、建立免疫检测方法奠定基础。
|
| 关 键 词： | 兽药 恩诺沙星 单链抗体 噬菌体展示技术 半抗原 |
| 收稿时间： | 2010/7/21 0:00:00 |
Construction and identification of anti-ENR phagedisplay scFv libraries |
| |
| WEN Kai,SHEN Jian-zhong,MENG Hui,WU Cong-ming,WANG Zhan-hui and ZHANG Su-xia.Construction and identification of anti-ENR phagedisplay scFv libraries[J].Journal of China Agricultural University,2011,16(2):118-124. |
| |
| Authors: | WEN Kai SHEN Jian-zhong MENG Hui WU Cong-ming WANG Zhan-hui and ZHANG Su-xia |
| |
| Institution: | WEN Kai,SHEN Jian-zhong,MENG Hui,WANG Zhan-hui,ZHANG Su-xia(College of Veterinary Medicine,China Agricultural University,Beijing 100193,China) |
| |
| Abstract: | This study focuses on construction and identification of immunized phage display library from splenocytes of hyperimmunized BALB/C mice for screening and isolation of scFv fragment against ENR,an alternative method to produce antibodies for veterinary drug residues detection.The total RNA was isolated from splenocytes of a BALB/C mouse hyperimmunized with the Enrofloxacin conjugated to chicken OVA.Variable light and heavy domains of the immunoglobulin genes were amplified by PCR and assembled to produce ful... |
| |
| Keywords: | veterinary drug enrofloxacin single chain antibody phage display techniques hapten |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《中国农业大学学报》浏览原始摘要信息 |
| 点击此处可从《中国农业大学学报》下载免费的PDF全文 |
|
