犬副流感病毒单克隆抗体的研制及其特性鉴定

为研制犬副流感特异性诊断试剂,我们以犬副流感病毒(CPIV)免疫8周龄BALB/c小鼠,采用淋巴细胞杂交瘤技术获得4株稳定分泌针对CPIV的单克隆抗体(MAb)细胞株,分别命名为4F386、584C9、4G7F4和4C9D8.4株MAb腹水针对CPIV的间接ELISA抗体效价达1:10~5～1:10~6,与犬瘟热病毒(CDV)和犬细小病毒(CPV)均不发生交叉反应.MAb 4F386和4C9D8为IgG,5B4C9和4G7F4为IgM.Western blot检测表明,4F386与CPIV的F蛋白发生特异性反应,4G7F4与CPW的HN蛋白发生特异性反应,而584C9和4C9D8不与变性的CPIV蛋白发生反应.4株MAb均具有中和病毒活性,间接免疫荧光检测均呈为阳性.本研究为进一步研制CPIV特异性诊断和治疗制剂创造了条件.

Production and characterization of the monoclonal antibodies against canine parainfluenza virus

Four monoclonal antibodies (MAb) against canine parainfluenza virus (CPIV), 4F3B6, 5B4C9, 4G7F4 and 4C9D8, were produced by fusing mice myeloma cell line SP2/0 with spleen lymphocytes from BALB/c mice immunized with CPFV yz0506 strain. Indirect ELISA assay showed that the antibodies had an ascite titre of 1:10~5-1 :10~6. All MAbs were highly specific and showed no reaction with Canine distemper virus (CDV) and Canine parvovirus (CPV). The 4F3B6 was specific to CPIV F protein, and 4G7F4 specific to CPIV HN protein, however neither 5B4C9 nor 4C9D8 could react with denatured CPIV structure protein in western blot. The MAbs had neutralization activity to CPIV and was shown to positively react with the MDCK cells infected with CPIV yz0506 strain. MAbs reported here could provide valuable tools for further development specific diagnosis and treatment reagent to CPIV infection.