摘要
目的研制羊布鲁菌O链M纯化抗原及其单克隆抗体。方法采用冷酚法提纯羊布鲁菌16M的O链抗原,并经琼脂糖凝胶免疫扩散和SDS-PAGE鉴定,用灭活的该抗原免疫BALB/c小鼠,制备单克隆抗体,并进行鉴定。结果建立3株分泌单克隆抗体的细胞株2D10、3C6和1E10,ELISA效价分别为1.380、1.109和1.048,与大肠杆菌0:157、小肠结肠炎耶尔森菌0:9、鼠伤寒沙门菌和猪胸膜肺炎放线杆菌均无交叉反应。3C6和1E10与牛布鲁菌544A发生交叉反应,2D10的亲和常数达5.30×10~7M^(-1)。结论已制备出纯化的羊布鲁菌O链M抗原及其单抗。
Objective To prepare purified B. melitensis 0 chain M antigen and its specific monoclonal antibody（McAb）. Methods Purify the O chain from B. melitensis 16M by cold phenol extraction and identify by agarose gel immunodiffusion and SDS-PAGE. Immunize BALB/c mice with inactivated O chain antigen to prepare McAb. Results Three cell strains secreting McAb against O chain antigen, i. e. 2D10,3C6 and 1 El0, were established, and their ELISA titers were 1. 380,1. 109 and 1. 048 respectively. The secreted McAbs showed no cross reaction with E. coli O：157, Y. enterocolitica 0：9 ,S. typhimurium and Actinobaciilus. However, the McAbs secreted by 3C6 and 1El0 showed cross reaction with B. abortus 544A,and the affinity constant of McAb secreted by 2D10 reached 5.30 x 10^7 M^-1. Conclusion Purified B. melitensis 0 chain M and its specific McAb were successfully prepared.
出处
《中国生物制品学杂志》
CAS
CSCD
2007年第10期771-774,共4页
Chinese Journal of Biologicals
关键词
羊布鲁菌
O链
M抗原
单克隆抗体
B. melitensis
0 chain
M antigen
Monoclonal antibody
