中国兽医科学2017，Vol.47Issue(6)：721-726,6.DOI:10.16656/j.issn.1673-4696.2017.06.008
鸡IFN-γ单克隆抗体的制备及其识别区的鉴定
Development of monoclonal antibodies against chicken IFN-γ (chIFN-γ) and identification of their recognizing regions
摘要
Abstract
Chicken IFN-γ(chIFN-γ) plays an important role in host.To develop monoclonal antibodies (McAbs)against chIFN-γ,chIFN-γ-His recombinant protein was used to vaccinate BALB/c mice three,of which the spleen cells were fused with the separated splenocytes with myeloma (SP2/0)cells,cloned,cultured and screened.4 hybridoma cell lines that stably secreted McAbs against chIFN-γwere screened and named as 2A10,2B10,4D10 and 4F9,respectively.The dissociation constant(Kd)of these McAbs were determined and their Kds were 1.06 × 10-10,7.35 × 10-10,1.10 × 10-10and 9.34 × 10-10,respectively.The isotypes of these McAbs were IgG2a,IgG1,IgG1 and IgG2b,respectively.Western-blot analysis demonstrated that chIFN-γ was recognized by 2A10,2B10,4D10,4F9 and targeted between amino acids 47 to 100,101 to 145 and 1 to 46,respectively,but not by chIL-2,chIL-4 and chIL-10.These McAbs would help to analyze the biological activities of chIFN-γ.关键词
鸡IFN-γ/原核表达/单克隆抗体Key words
chicken IFN-γ/prokaryotic expression/monoclonal antibody分类
农业科技引用本文复制引用
柳亚楠,王永强,李晓齐,曹红,郑世军..鸡IFN-γ单克隆抗体的制备及其识别区的鉴定[J].中国兽医科学,2017,47(6):721-726,6.基金项目
国家自然科学基金项目(31272543 （）
31430085) （）
现代农业产业技术体系建设专项(NYCYTX-41) （NYCYTX-41）
