鸭疫里默氏杆菌脂多糖特异性单克隆抗体的制备及鉴定
Preparation and Identification of Specific Monoclonal Antibody Against Lipopolysaccharide of Riemerella anatipestifer
武梦思 1陈美潼 2朱悦 2罗廷荣 3丁铲 2于圣青4
作者信息
- 1. 广西大学动物科学技术学院,广西 南宁 530004;中国农业科学院上海兽医研究所,上海 200241
- 2. 中国农业科学院上海兽医研究所,上海 200241
- 3. 广西大学动物科学技术学院,广西 南宁 530004
- 4. 中国农业科学院上海兽医研究所,上海 200241;江苏农牧科技职业学院,江苏省兽用生物制药高技术研究重点实验室,江苏 泰州 225300
- 折叠
摘要
为制备鸭疫里默氏杆菌脂多糖特异性单克隆抗体,研究选择SDJ-80菌株经过灭活处理后的全菌作为抗原,对小鼠进行免疫接种,取免疫小鼠脾细胞与SP2/0细胞进行细胞融合;细菌脂多糖包被酶标板,间接ELISA法检测细胞上清,获得抗体阳性细胞并进行亚克隆,将成功获得的阳性杂交瘤细胞株连续传10代,并进行抗体效价和特异性检测.结果显示:成功获得一株单克隆抗体1E6,制备的腹水单克隆抗体效价高于1∶204 800,该单克隆抗体与血清1、2、6、7和10型鸭疫里默氏杆菌均能发生特异性反应.研究表明,获得的单克隆抗体1E6为针对鸭疫里默氏杆菌脂多糖的特异性单克隆抗体,可用于鸭疫里默氏杆菌脂多糖分子结构和诊断新方法的研究.
Abstract
In order to prepare specific monoclonal antibody(mAb)targeting the lipopolysaccharide(LPS)of Riemerella anatipestifer(RA),BALB/c mice were vaccinated with inactivated bacteria of RA SDJ-80 strain,and the mAb specific to LPS was prepared by fusing myeloma cells SP2/0 with immunized positive mouse spleen cells.The LPS was used as the coating antigen,and positive clones were screened using indirect ELISA method,then subjected to sub-cloning.The prepared positive hybridoma cell line was continuous passaged for 10 generations,and the titer and specificity of the mAb were detected.The results showed that a mAb named as 1E6 was obtained successfully,the titer of which was higher than 1:204 800 and could react specifically with RA serotype 1,2,6,7,and 10 strains,which could be used for future study on RA LPS structure and novel diagnostic methods.
关键词
鸭疫里默氏杆菌/脂多糖/单克隆抗体Key words
Riemerella anatipestifer/lipopolysaccharide/monoclonal antibody引用本文复制引用
出版年
2025
NETL
NSTL
万方数据