猪SIgA分泌片的分离纯化及其多克隆抗体的制备  

Separation and Purification of Secretory Component of Porcine SIgA and Preparation of Polyclonal Antibodies against Secretory Component

作　　者：张辉[1] 崔焕忠[1] 杨欢[1] 马思慧[1] 吴天成[1] 兰海楠[1] 郑鑫[1] 

机构地区：[1]吉林农业大学动物科技学院,长春130118

出　　处：《中国兽药杂志》2014年第2期20-23,共4页Chinese Journal of Veterinary Drug

基　　金：吉林农业大学博士启动基金项目(201221);吉林省科技厅科技引导计划国际科技合作项目(0130413037GH)

摘　　要：为制备抗分泌片多克隆抗体,采用分子筛凝胶层析、离子交换层析和硫酸铵盐析法从猪初乳中分离纯化分泌片和SIgA。SDS-PAGE鉴定表明,纯化的SIgA呈现分泌片、重链和轻链三条带,大小约为70 ku;凝胶薄层扫描分析SIgA和分泌片纯度分别为90%和89%;琼脂扩散鉴定表明,SIgA和分泌片与抗分泌片单克隆抗体发生了特异反应。以纯化分泌片为抗原,制备抗分泌片多克隆抗体,琼脂扩散检测多抗的效价为1∶32。高纯度分泌片的提取及高效价抗分泌片多抗的制备为粘膜免疫研究奠定了物质基础。In order to prepare polyclonal antibodies against secretory component（SC） ,porcine SIgA and SC were purified from colostrums by a procedure of chromatography with Sephadex G-200, ion-exchange chromatography with DEAE 52 and precipitation with saturated ammonium sulfate. Identification by SDS-PAGE showed that the SIgA took on three straps： SC, heavy chain and light chain. And the molecular weight of the purified SC was about 70 Ku. Gel TLC analysis showed that the purities of the SIgA and secretory were 90% and 89% respectively. Identification by agar double immunodiffusion showed that the SIgA and SC reacted with the anti-SC McAb and the titer of polyclonal antibody was 1 ： 32. The SC of high purity and the polyclonal antibody of high titer were prepared. This study laid a solid material foundation for the research of mucosal immunity.

关 键 词：猪 分泌片 纯化 多克隆抗体 

分 类 号：S852.4[农业科学—基础兽医学]