抗伪狂犬病毒Ea株单克隆抗体的制备及鉴定

Preparation and Characterization of Monoclonal Antibody to PRV Ea Strain

将伪狂犬病毒Ea株纯化后免疫雌性Balb/c小鼠,取其脾细胞与SP2/O的骨髓瘤细胞融合,用间接酶联免疫吸附试验(ELISA)进行筛选,获得了2株抗PRV的单克隆抗体的杂交瘤细胞,分别命名为3G7E3和4A6F5,其细胞培养上清及小鼠腹水效价(ELISA)分别为1:212、1:212及1:100×212、1:100×212。特异性试验和免疫荧光结果显示,3G7E3和4A6F5仅与PRV反应,而与PPV、PRRSV、JEV和HCV等病毒不发生交叉反应;用单抗建立的夹心ELISA试验能检出伪狂犬病毒。表明本研究所获得的2株单抗是PRV特异性的。这2株分泌抗PRV单抗的杂交瘤细胞株的获得为进一步建立准确快速的抗原检测方法奠定了基础。

After immunization of Balb/c mice with purified PRV, Two hybridoma cell strains a-gainst PRV named 3G7E3 and 4A6F5, respectively, were developed after fusion between SP2/0 myeloma cells and the stimulated splenocytes. The indirect enzyme-linked immunosorbent assay (ELISA) titer of culture supernatant and ascites were 1: 212, 1: 212 and 1 :100 ×212, 1: 100×212. No cross reaction was found when Mabs reacted JEV, PRRSV, PPV and HCV. The specific reactivity of Mabs with PRV was confirmed by IFA and Mab-based sandwich ELISA. These results indicated that we established the specific hybridoma cells secreting Mab against PRV and Mabs could be used to develop diagnostic methods for detection of PrV antigen as well as to investigate protein functions of PRV .