鲤春病毒血症病毒单克隆抗体制备及鉴定

Preparation and identification of monoclonal antibody against spring viraemia of carp virus

为制备鲤春病毒血症病毒(SVCV)单克隆抗体(MAb),本研究用纯化的SVCV免疫BALB/c鼠,通过杂交瘤细胞技术,制备2株抗SVCV的MAb,分别命名为2A3和3H7。经间接ELISA检测腹水效价为1:10240和1:12150。亚类鉴定证实,这2株亚类均为IgG1型。经非竞争酶免疫试验测定2A3和3H7的抗体亲和力常数分别为4.86×108L/mol和1.86×109L/mol,3H7相对亲和力高于2A3。MAb 2A3和3H7的饱和度均为1:400,叠加试验所得增值指数为63.2%,大于50%。抗体反应增值结果表明:两株MAb分别针对不同抗原位点。免疫印迹分析检测表明,两株纯化的MAb均可与SVCV抗原发生特异反应。

Monoclonal antibodies（MAb） against spring viraemia of carp virus（SVCV） were prepared by BALB/c mice immunization and cell fusion techniques.Two MAbs were obtained with titers from 1：10,240 to 1：12,150 by ELISA.Antibody affinities were 4.86×108 L/mol and 1.86×109 L/mol,respectively.Western blot and competitive assay showed that the MAbs reacted with the same protein of SVCV,but against different kinds of epitopes.These MAbs provided diagnosis reagent for SVCV detection method establishment.