摘要
原料羊乳中掺入牛乳会对其制品的食品安全造成损害.本研究的目的是建立一种免疫层析试纸条的检测方法检测羊乳原料乳的掺假.以牛乳αs1-CN及牛乳β-LG为检测对象,制备两者的特异性抗体,建立双联胶体金免疫层析试纸条.该实验制备的多克隆抗体α-CN IgG和β-LG IgG蛋白质浓度分别为4.86和5.99 mg/mL,两种抗体的效价为1∶25 600和1∶204 800,制备的胶体金溶液最适pH值为8.0,最佳蛋白标记量为10μL,且β-LG IgG中α-CN IgG的最适掺入量为20%.试纸条以玻璃纤维素膜VL68作为金标垫、硝酸纤维素膜SarteriesCN 95作为NC膜,分别用6μL,1 mg/mL的α-CN IgG,6μL,1 mg/mL的β-LG IgG包被NC膜T线、用6μL,1mg/mL二抗包被C线.该试纸条可特异性的检测出α-CN和β-LG,无明显的交叉反应,且可检出的牛乳最低掺入量为5%.
Aduteration of raw bovine milk to goat milk causes damage to food safety of its products,especially to consumers with bovine milk allergy.The purpose of this study was to establish a method of immunochromatographic strips to detect adulteration of raw goat milk.Bovine milkαs1-CN（αs1-Casein）and milkβ-LG（β-Lactoglobulin）was taking as the test object to develop double the colloidal gold immunochromatographic strip.Their specific ployclonal antibodies was prepared.The protein concentrations of polyclonal antibodies ofα-CN IgG andβ-LG IgG were 4.86 mg/mL and 5.99 mg/mL and the titers of these two antibodies were 1∶25 600 and 1∶204 800 respectively.In the preparation experiments of colloidal gold,the optimal pH was 8.0,and the best protein labeling amount was 10μL,and the optimal incorporation ratio ofα-CN IgG inβ-LG IgG was 20%.Test strips were coated with glass fiber VL68 as gold standard and Sarteries CN 95 as nitrocellulose NC membranes.NC membrane was coated with 6μL,1.0 mg/mLα-CN IgG and 6μL,1.0 mg/mLβ-LG IgG on T lines,and use 6μL,1.0 mg/mL secondary antibody to coated C line as well.The test strip can detectα-CN andβ-LG with no obvious cross-reaction,and the lowest detectable content of milk is5%.
作者
薛海燕
许淼
白文卿
张颖
薛丽欢
XUE Hai-yan;XU Miao;BAI Wen-qing;ZHANG Ying;XUE Li-huan(School of Food and Biological Engineering, Shaanxi University of Science ＆ Technology, Xi＇an 710021, China)
出处
《陕西科技大学学报》
CAS
2018年第3期46-51,62,共7页
Journal of Shaanxi University of Science & Technology
基金
国家自然科学基金项目(31301405)
陕西省科技厅科技统筹创新工程计划项目(2013KTZB02-02-05(2))
陕西省大学生创新创业训练计划项目(1344)
关键词
牛羊乳掺假
快速检测
胶体金免疫层析
adulteration of goat milk with bovine milk
rapid detection
colloidal gold immunochromatography
