猪瘟病毒野毒株E2蛋白特异性单抗免疫抑制制备法的建立

Preparation of a monoclonal antibody against E2 protein of CSFV

采用环磷酰胺免疫抑制法制备特异性识别猪瘟病毒(CSFV)野毒株的单克隆抗体,用纯化的杆状病毒表达的重组HCLV-E2蛋白作为耐受原,Shimen-E2蛋白作为免疫原,免疫BALB/c小鼠,取其脾淋巴细胞与SP2/0骨髓瘤细胞融合,获得了1株能稳定分泌抗E2蛋白单克隆抗体的杂交瘤细胞株,结果显示,该单克隆抗体能够与CSFV石门株发生反应,同时能与1.1、2.1、2.2和2.3基因亚群CSFV野毒发生特异性反应,并具有中和活性,而与CSFV兔化弱毒疫苗株不发生反应。表明,该单克隆抗体可以用于鉴别CSFV野毒株和兔化弱毒疫苗株。

To preparate a monoclonal antibody（McAb） specific to the E2 protein of classical swine fever virus （CSFV） wild-type, BALB/c mice were immunized with the purified recombinant baculovirusexpressing E2 protein from C-strain（HCI.V E2） as tolerogen and with E2 protein from Shimen strain （Shimen-E2） as immunogen. The immunized murine splenic lymphocytes were fused with SP2/0 hybridoma cells. One myeloma cell line stably secreting McAb against E2 protein was obtained. The McAb could recognize CSFV Shimen strain,but not to C-strain,and could react with several subgroups,including 1.1, 2.1,2.2 and 2.3 of CSFV wild-type by the neutralizing immunofluorescence test. The results showed that the McAb could be used to differentiate CSFV wild-type strains from C-strain.