志贺样毒素Ⅱ型变异体B亚基单克隆抗体的制备  

Preparation of monoclonal antibody against Shiga-like toxin typeⅡvariant B

作　　者：丁琳[1] 师东方[1] 付海兵[1] 于迪[1] 杨旭东[1] 单安山[2] 

机构地区：[1]东北农业大学动物医学学院,黑龙江哈尔滨150030 [2]东北农业大学动物科学技术学院,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2009年第3期222-225,240,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：东北农业大学"211"工程人才基金;黑龙江省博士后研究基金;黑龙江省"十五"重点攻关项目(GB05B501);东北农业大学创新团队项目(CX2008-3)

摘　　要：为建立针对仔猪水肿病SLT-Ⅱe毒素的检测方法,应用PCR技术克隆SLT-ⅡeB基因,并将其克隆到原核表达载体pET-30a进行表达。以纯化的融合蛋白免疫BALB/c小鼠,并利用淋巴细胞杂交瘤技术获得6株稳定分泌抗SLT-ⅡeB的单克隆抗体(McAb)细胞株,其抗体亚类包括IgM、IgA、IgG2b和IgG2a,而且轻链均为κ链。阻断ELISA结果显示,6株McAb均能特异性识别天然SLT-Ⅱe毒素。相加ELISA结果显示,3G7和4F3两株是针对不同抗原表位的McAb。To establish a new detection method for Shiga-like toxin type Ⅱ variant （SLT-Ⅱ e）, the SLT-Ⅱ e B gene was amplified by PCR and aequence compared with reference strains. The gene was subcloned into pET-30a vector for expression and the recombinant product of SLT-Ⅱ e B was used to immunize the BALB/c mice. Six hybridoma cell lines secreting monoclonal antibodies （McAbs） against SLT-Ⅱe B were established, The McAbs belonged to IgM, IgA, IgG2b and IgG2a subtype with κ chain. Blocking ELISA confirmed that all McAbs were able to reacted with native SLT-Ⅱ e. Addition ELISA confirmed that McAbs 3G7 and 4F3 recognized different epitopes.

关 键 词：仔猪水肿病 志贺样毒素Ⅱ型变异体B亚基(SLT-ⅡeB) 单克隆抗体 

分 类 号：S852.613[农业科学—基础兽医学]