摘要
以单核细胞增生李斯特菌细胞碎片免疫BALB/c小鼠,间接ELISA法成功筛选获得2株稳定分泌抗LM的单克隆杂交瘤细胞株4A7、4H11。抗体效价为1∶160 000以及1∶20 000,亚型为IgG1、IgG2a,Dot-ELISA结果表明4A7和4H11单克隆抗体具有很好的属特异性,Western blot分析表明4A7、4H11抗体分别与单核细胞增生李斯特菌62 kDa以及32 kDa外膜蛋白抗原表位结合,胶体金免疫电镜实验进一步确证以上抗体可有效识别单核细胞增生李斯特菌细胞表面抗原。
Two strains of mAbs against L. monocytogenes, namely 4A7 and 4Hll, are generated by immuned the BALB/c mice using the cell fragments of L. monocytogenes as the immunogen. The titers of mAbs are 1：160 000, 1：20 000 and the subclasses of mAbs are IgG1, IgG2a, respectively. The specificity of the mAbs is evaluated using Dot-ELISA method and the two strains of mAbs show a high infinity to Listeria monocytogenes, Listeria ivanovii, Listeria welshimeri, Listeria seeligeri and Listeria innocua whereas exhibit no cross-reactivity with Listeria grayi and other 30 common bacterias. Western-blot analysis showes the mAbs 4A7 and 4Hllcan recognize the membrane proteins of L. monocytogenes with the molecular weight of 62 kDa and 32 kDa, respectively. Immuno-gold based transmission electron microscopy further demonstrates that the mAbs are capable of binding the membrane antigens of L. monocytogenes.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2013年第5期56-61,共6页
China Biotechnology
基金
"十二五"国家科技支撑计划(2011BAK10B06
2011BAK10B01)
国家自然科学基金(31271863)
江西省科技厅"井岗之星"人才计划(2010DQ00800)资助项目
