摘要
利用纯化的人载脂蛋白Ｅ（ａｐｏＥ）作为抗原，常规免疫纯系大耳白家兔，获得高效价、高特异性的抗血清，抗体滴度经ＥＬＩＳＡ法检测达１：１２８００倍稀释度，抗体特异性检验证明抗体只与ａｐｏＥ反应。抗血清经ＰｒｏｔｅｉｎＡ－ＳｅｐｈａｒｏｓｅＣＬ－４Ｂ亲和层析纯化，分离为碱性洗脱峰Ⅰ和酸性洗脱峰Ⅱ，峰Ⅱ经ＳＤＳ－ＰＡＧＥ检测在分子量为２７ｋｄ和５４ｋｄ处呈现两条带，分别相当于抗体的重链和轻链的分子量，证明峰Ⅱ为纯化的抗体，这就为建立测定人血清ａｐｏＥ浓度的免疫学方法奠定了基础。
Apolipoprotein E was purified from human serum by ultracentrifugation and chromatography and used to immunize rabbits,The rabbit antiserum obtained was shown to be high specifity to human apolipoprotein E by ELISA and purified by affinity column on protein A-Sepharose CL4B.Two peaks were eluted from the column and the second peak was proved to be the purified IgG by SDS-PAGE.All the work above will lay the foundation for the determination of human apolipoprotein E with immunoassay.
出处
《天津医科大学学报》
1995年第3期1-3,共3页
Journal of Tianjin Medical University
基金
天津市二十一世纪青年科学基金
