重组人tumstatin的原核生物表达和多克隆抗体制备

目的原核生物表达可溶性的tumstatin抗原并制备相应的多克隆抗体。方法利用原核表达载体pMAL-tumstatin在大肠埃希菌BL21中表达tumstatin,经Amylose Resin亲和层析柱和Q Sepharose Fast Flow柱纯化tumstatin,以纯化蛋白免疫新西兰兔,获得抗tumstatin多克隆抗体。利用western blot和ELISA法对多克隆抗体进行特异性和效价检测。结果tumstatin蛋白表达成功。SDS-PAGE分析表明其为可溶性表达。成功获得了tumstatin纯品及兔抗tumstatin多克隆抗体。ELISA法表明多克隆抗体效价>5 000。western blot检测证明多克隆抗体的特异性良好。结论成功表达、纯化tumstatin蛋白,并获得高特异性、高效价兔抗tumstatin多克隆抗体,为其在临床免疫学检测应用奠定了基础。

Expression of recombinant human tumstatin in Escherichia coli and preparation of its polyclonal antibody

Objective To express human tumstatin in E.coli and to prepare antibody against tumstatin.Methods Tumstatin was expressed in BL21 with pMAL-c2 expression vector and purified with Amylose Resin column and Q Sepharose Fast Flow column.The purified tumstatin was used to immunize New Zealand rabbits for preparing polyclonal antibody.The specificity and reactivity of the antibody were evaluated by western blotting and ELISA.Results The tumstatin was highly expressed and purified.The polyclonal antibody against tu...