摘要
建立了检测小麦过敏原醇溶蛋白的定量ELISA抗体夹心法，以2μg／mL的鼠抗醇溶蛋白单抗包被酶标板，HRP标记兔抗醇溶蛋白抗体作为酶标抗体，工作浓度为1：8000，将醇溶蛋白精确配制成一定浓度的标准品，检测醇溶蛋白的含量。最佳的线性范围是19．53～5000ng／mL，R^2≥0．99，定量检测限为19．53ng／mL，对应的食品样品检测限为7．81ppm。样品的回收率在93．57%-105．72％之间，实验内CV为3．07％～5．91％，实验间CV为0．26％～5．24％，仅对大麦有一定的交叉反应，可在4℃放置两周。本法准确度和精密度高，重复性和稳定性好，特异性较强，适用于食品过敏原小麦醇溶蛋白的检测。
To establish a method for quantitative analysis of gliadin from wheat in foods by ELISA. The mouse anti-gliadin McAb was used as a coating antibody in the concentration of 2μg/mL,and a horseradish peroxidase- conjugated anti-gliadin as an enzyme-labeled antibody in the optimal working dilution of l：8000,and a series of concentrations of gliadin solution as standards.The contents of gliadin in samples were determined.The optimal linear range was 19.53-5000ng/mL and R^2 ≥0.99.The quantitative limit of detection was 19.53ng/mL,which was corresponding to 7.81ppm in food samples.The recovery rate for the accuracy test was 93.57%-105.72%.The coefficient of variation for precision assay were 3.07%-5.91% in the same test and 0.26%-5.24% among three tests.Cross reaction was only observed with barley.The set of kit could be placed 4℃ for two weeks.The method is sensitive,accurate,specific.It is suitable for quantitative determination of gliadin in foods.
出处
《食品工业科技》
CAS
CSCD
北大核心
2010年第11期360-362,365,共4页
Science and Technology of Food Industry
基金
科技部质检公益项目(2007GYJ036)
