Xiang Keyu, Cheng Yi, Zhou Meizhen, Sun Leilei, Ji Yikuan, Wang Yu, Zhang Baoshi, Luo Yongwen, Ju Chunmei
1 College of Veterinary Medicine, South China Agricultural University , Guangzhou, P.R. China .
Monoclon Antib Immunodiagn Immunother. 2014 Dec;33(6):409-13. doi: 10.1089/mab.2014.0046.
Early protein 0 (EP0) is especially important for modulating PRV gene expression and reactivation from the latent state, but the mechanisms have not been elucidated. In this study, six monoclonal antibodies (MAbs) against EP0 protein of PRV were generated and their characterizations were investigated. Western blot analysis showed all six MAbs could react with immunizing antigen, but only 2B12 and 2C6 could react with native EP0 protein from PRV-infected cells. ELISA additivity tests revealed that at least three epitopes in EP0 were defined by six MAbs. The epitope recognized by MAb 2B12 was further identified in 287-292 aa of EP0 protein using a series of expressed overlapping peptides. These MAbs may provide valuable tools for further research of the functions of EP0 in PRV infection.
早期蛋白0(EP0)对于调节伪狂犬病毒(PRV)基因表达及从潜伏状态重新激活尤为重要,但其机制尚未阐明。在本研究中,制备了六种针对PRV的EP0蛋白的单克隆抗体(MAb),并对其特性进行了研究。蛋白质免疫印迹分析表明,所有六种单克隆抗体均能与免疫抗原发生反应,但只有2B12和2C6能与PRV感染细胞中的天然EP0蛋白发生反应。酶联免疫吸附测定(ELISA)加和试验表明,六种单克隆抗体确定了EP0中至少三个表位。使用一系列表达的重叠肽,在EP0蛋白的287 - 292个氨基酸中进一步鉴定了单克隆抗体2B12识别的表位。这些单克隆抗体可能为进一步研究EP0在PRV感染中的功能提供有价值的工具。