中国动物传染病学报2020，Vol.28Issue(4)：52-57,6.
猪流行性腹泻病毒N蛋白的原核表达及其单克隆抗体的制备
PROKARYOTIC EXPRESSION OF PORCINE EPIDEMIC DIARRHEA VIRUS N PROTEIN AND PREPARATION OF MONOCLONALANTIBODIES
摘要
Abstract
In order to obtain nucleocapsid (N) protein of porcine epidemic diarrhea virus (PEDV) and prepare monoclonal antibodies (McAb), the recombinant plasmid pET28a-N was constructed and expressed in BL21(DE3) competent cells after induced with IPTG. Then, the recombinant N protein was purified by Ni-NTA affinity and gel filtration chromatography and used to immunize BALB/c mice. The splenocytes from the immunized mice were fused with SP2/0 myeloma cells and the hybridomas were screen for secretion of MAbs by indirect ELISA. As a result, 2 hybridoma cell lines (1A3 and 4F6) that stably produced anti-N protein antibodies were obtained and further analyzed. The antibody subtypes of both McAbs were IgG1. The antibody titers of 1A3 and 4F6 in ascites were 5.12×105 and 1.28× 105 as determined in indirect ELISA, respectively. This study laid an important foundation for the functional research of N protein and development of diagnostic kits of PEDV.关键词
猪流行性腹泻病毒/N蛋白/原核表达/单克隆抗体Key words
Porcine epidemic diarrhea virus/ N protein/ prokaryotic expression/ monoclonal antibody分类
农业科技引用本文复制引用
李任峰1,田香勤2,韩笑1,王晓斐1,姜金庆1,夏小静1,王异民1,王自良1..猪流行性腹泻病毒N蛋白的原核表达及其单克隆抗体的制备[J].中国动物传染病学报,2020,28(4):52-57,6.基金项目
河南省科技攻关项目(192102110190) （192102110190）
河南省科技攻关项目(202102110255) （202102110255）
河南省高校科技创新团队计划(20IRTSTHN025) （20IRTSTHN025）
