《Lipocalin核糖体展示文库的构建及雌二醇特异性模拟抗体的筛选 Construction of Ribosome Displayed Library Based on the Lipocalin Scaffold and Screening Anticalins to Estradiol》.pdfVIP

ofAnhui 责任编辑石金友责任校对况玲玲 安徽农业科学，Journal Agi．Sci．2013，41(31)：12251—12254，12261 Lipocalin核糖体展示文库的构建及雌二醇特异性模拟抗体的筛选 刘建青1，宁保安2，高志贤2+ (1．包头师范学院生物科学与技术学院，内蒙古包头014030；2．天津市卫生学环境医学研究所，天津300050) 糖体一mRNA三联复合体文库。以小分子抗原雌二醇为靶标，采用固相亲和方法筛选抗雌二醇抗体，通过改变Mg“浓度将模拟抗体一 mmol／L 行下一轮淘筛。[结果]通过5轮生物淘筛，模拟抗体得到富集，获得高特异性抗雌二醇抗体。其中有一株克隆的亲和力为54 (Kon=4．9756×104+／M·S；K0ff=0．0027+／s)。，c。与LOD值分别为50和o．071ng／“。[结论]获得的雌二醇模拟抗体可以作为 抗体并用于检测动物体内雌二醇的残留。 关键词雌二醇；BBP；核糖体展示；突变文库 中图分类号S188文献标识码A 文章编号0517—6611(2013)31—12251一04 ConstructionofRibosome BasedOilthe Scaffoldand AnlkalinstoEstradiol DisplayedLibrary LipocaUn Screening LIU al of Scienceand Normal Jhn-qi赡et(SchoolBiological Technology，BaotouUniversity，Baotou，InnerMongolia014030) Abstract ribosome andscltenoutestradiol simulation Method】The 『Objective]Toconstruct“pocalindisplaylibrary specific antibody．I memberofthe fromPieris asamodel forthe of brassicae，was bilin—bindingprotein(BBP)，alipecalinfamily employed system preparation arandom orderto steroid witllextensivemedicalfunctiontoanimalsand library．In specificallycomplexestradiol(E，)，aligand poultries，16 aminoacidresiduesattllecenterofthe iSformedfour on ofan to bindingsite，which by loopstop eight—stranded8一barrel，weresubjected random the the BBP targeted mutagenesis．Fromresultinglibraryestradiol—binding bindingactivity for like beenselectedcombineduseofribosomeand ResultFourvariantswere compoundsestradi01．have by displayscreeningtechniques．I1 identifiedthat estradiolwith estradiol th