利用针对人IgE结合表位的特异性多克隆抗体制备夹心ELISA检测牛β-乳球蛋白致敏残基
Development of sandwich ELISA for testing bovine β-lactoglobulin allergenic residues by specific polyclonal antibody against human IgE binding epitopes.
作者信息
He Shengfa, Li Xin, Gao Jinyan, Tong Ping, Chen Hongbing
机构信息
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China; School of Food Science & Technology, Nanchang University, Nanchang 330031, PR China; Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047, PR China.
State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, PR China; School of Food Science & Technology, Nanchang University, Nanchang 330031, PR China.
出版信息
Food Chem. 2017 Jul 15;227:33-40. doi: 10.1016/j.foodchem.2017.01.060. Epub 2017 Jan 16.
Bovine β-lactoglobulin (BLG) is the main allergen in cows' milk, and the most commonly used method for detecting BLG is enzyme-linked immunosorbent assay (ELISA). However, antibodies used in commercial ELISA kits do not recognize specifically BLG IgE epitopes. Here, an antibody specific to IgE linear epitopes for BLG was used to develop a sandwich ELISA using a rabbit anti-BLG polyclonal antibody. The linear range for BLG detection was 31.25-8000ng/mL and limit of detection was 1.96ng/mL. BLG content in dairy samples was determined, and there was a good agreement between this immunoassay and reversed-phase high-performance liquid chromatography with high recovery. Additionally, BLG content in food samples had an average recovery of 104.25%. Allergenic residues were also detected in hydrolyzed infant formulas. The method developed could be a practical approach to determine BLG and its allergenic residues in food with a high degree of sensitivity, reliability and recovery.
牛β-乳球蛋白（BLG）是牛奶中的主要过敏原，检测BLG最常用的方法是酶联免疫吸附测定（ELISA）。然而，商业ELISA试剂盒中使用的抗体不能特异性识别BLG IgE表位。在此，使用针对BLG IgE线性表位的特异性抗体，利用兔抗BLG多克隆抗体制备夹心ELISA。BLG检测的线性范围为31.25-8000ng/mL，检测限为1.96ng/mL。测定了乳制品样品中的BLG含量，该免疫测定法与反相高效液相色谱法之间具有良好的一致性，回收率高。此外，食品样品中BLG含量的平均回收率为104.25%。在水解婴儿配方奶粉中也检测到了致敏残留物。所开发的方法可能是一种实用的方法，能够高度灵敏、可靠且高回收率地测定食品中的BLG及其致敏残留物。
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