酶联免疫吸附法与二喹啉甲酸法检测核桃蛋白浓度的比较研究

Comparison of enzyme-linked immunosorbent assay and bicinchoninic acid assay for detection of walnut protein concentration in food

中文关键词:  间接竞争ELISA法  BCA试剂盒  蛋白浓度检测

英文关键词:indirect enzyme-linked immunosorbent assay  bicinchoninic acid kit  protein concentration detection

中文摘要:

      目的 探究间接酶联免疫吸附法(enzyme-linked immunosorbent assay, ELISA)和二喹啉甲酸 (bicinchoninic acid, BCA)试剂盒2种方法在核桃蛋白浓度检测中的优缺点和适用范围。方法 对本实验室提取的核桃蛋白进行烘烤、高温高压以及微波加热处理后, 用ELISA法和BCA法2种方法分别检测其蛋白质浓度。结果 在不同的热加工处理条件下, ELISA对蛋白浓度测定结果存在显著性差异, 高度依赖热处理的类型。与之相比, 使用BCA法测定蛋白浓度时, 受到热加工处理影响较小。结果提示, 在食物加工中的一些剧烈条件下, 蛋白质的高级结构很容易遭到破坏, 蛋白质结构的改变会干扰其与抗体结合部位的作用, 导致ELISA法测定的结果不准确。BCA法测定蛋白质浓度的准确性较高, 且受加热影响较小。结论 在剧烈的食品加工条件下, 尤其当这种条件引起蛋白结构发生改变后, 对其蛋白质浓度进行测定时, 可选用BCA法进行测定。

英文摘要:

      Objective To investigate the advantages, disadvantages and application range of indirect enzyme-linked immunosorbent assay (ELISA) and bicinchoninic acid (BCA) kits in determination of walnut protein concentration. Methods After baking, high temperature, high pressure and microwave heating, the walnut protein extracted in our laboratory was tested by ELISA method and BCA method. Results There were significant differences in the results of protein concentration detected by ELISA under different thermal processing conditions and it highly dependents on the type of heat treatment. In contrast, the BCA method used to determine the protein concentration had less affected by thermal processing. The results suggested that the high-level structure of the protein could easily be destroyed under severe conditions in food processing. The change of protein structure could interfere with the binding site of antibody, resulting in the inaccurate result of ELISA. It was of high accuracy of utilizing BCA to determine the protein concentration and it was less affected by the heating. Conclusion In the severe food processing conditions, especially when the conditions resulted in changes in the protein structure, the protein concentration can be measured by BCA method.