摘要
以纯化的酵母重组表达的犬细小病毒VP2单位免疫BALB/C小鼠,采用B淋巴细胞杂交瘤技术,通过ELISA方法筛选,获得4株能稳定分泌抗犬细小病毒CPV结构蛋白VP2的单克隆抗体杂交瘤细胞株。4株单克隆抗体中,2株属于IgG2b亚类,2株属于IgG1亚类,其腹水效价可达到1∶51 200和1∶204 800,细胞培养上清液效价可达1∶256、1∶512。ELISA分析表明,这些单抗仅与CPV及其VP2发生特异性反应,而与CDV和CAV-1及CAV-2没有交叉反应;荧光免疫染色病毒检测进一步表明单克隆抗体的特异性效果好。这些特异性单抗的制备为建立有效的检测犬细小病毒感染奠定了基础。
Monoclonal antibodies （McAb） against canine parvovirus （CPV） were produced by fu- sing myeloma cell line SP2/0 with spleen cells from Balb/C mice immunized with purified recom- binant VP2 protein expressed in yeast Pichia pastoris. Four hybridoma cell lines secreting specific antibodies against CPV were identified by screening hybridoma culture supernatants with indirect ELISA assay. The identified subtypes of four monoclonal antibadies were 2 as IgG2b and 2 as IgG1. Their titre in culture supernatants and ascitic fluids ranged from 1：51 200 to 1 ： 204 800 and 1： 256 to 1 ： 512, respectively. ELISA analysis showed that those McAbs specifically reconga- nized antigenic epitopes of CPV or VP2,but not those of onther canine virus such as CDV or CAV. The immun-staining CPV infectious cells using the four McAbs confirmed their specificity to CPV. The four specific monoclonal antibodies will provide a basis for an efficient test mentods for CPV infection.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2012年第11期1629-1633,共5页
Chinese Journal of Veterinary Science
基金
石家庄市科技支撑计划(09150293A)
