双抗体夹心ELISA定量检测牛乳中酪蛋白方法的建立

Development of sandwich-antibody enzyme linked immunosorbent assay for the rapid detection of casein in milk

实验使用酪蛋白标品为抗原,免疫新西兰大白兔制备抗酪蛋白的多克隆抗体,并免疫BALB/C小鼠制备抗酪蛋白单克隆抗体,以单克隆抗体包被,HRP标记多抗,成功地建立了检测牛乳中酪蛋白的双抗体夹心ELISA法,标准曲线在6～1666 ng/mL范围内线性良好,r2=0.9949,最低检出限为7 ng/mL,具有很好的特异性。通过建立双抗体夹心ELISA法测定酪蛋白含量,可为牛乳掺假和食物过敏原的检测及标签管理提供理论及技术基础。

Casein was used as the antigen to immune the New Zealand rabbits and BALB/C mice respectively in the experiment.The anti-casein polyclonal antibodies and monoclonal antibodies were prepared.The monoclonal antibody was used as the coating antibody,and HRP-polyclonal antibody was retained.The sandwich ELISA to detect the casein of milk was developed successfully,which had a standard curve in the 6～ 1666 ng/mL range of good linearity with r2=0.9949.The detection limit of the method was 7 ng/mL.The method could be use as a theory and technological base of casein detection of milk adulteration and food allergen test.