Analysis of the amino-carbonyl complexes in commercial milk products affected milk allergy

对名古屋市幼儿园和其他地方的1514名儿童进行了食物过敏调查。患病比例为10%，40.6%的患者在1岁之前患病。鸡蛋和奶制品是他们的主要过敏原。不同患儿的断奶饮食和哺乳方式无明显差异。在有过敏儿童的家庭中，更多的家庭成员易患过敏症。针对婴儿过敏的母亲在孕期和哺乳期对食物的喜好，本研究利用该单抗对脱脂奶、婴幼儿奶粉、巴氏杀菌奶和乳饮料等商品奶制品中潜在的乳糖蛋白美拉德加合物进行了特异性检测。用酶联免疫吸附试验在所有8个婴儿奶样中检测到Meilard加合物，但在原料奶中检测不到。蛋白质印迹分析表明，乳铁蛋白、牛血清白蛋白、免疫球蛋白等高分子量乳蛋白以及婴儿乳中的酪蛋白与该单抗有较强的染色。在6个脱脂牛奶样品中也检测到乳糖-蛋白质美拉德加合物。在脱脂牛奶的情况下，在低分子牛奶蛋白如β-LG和α-LA以及酪蛋白中观察到了高反应性的单抗。对于巴氏杀菌牛奶，在LLT牛奶中几乎没有检测到美拉德加合物，因此该单抗可用于乳糖-蛋白质氨基甲酰加合物的特异性检测和食品和食品复杂混合体系中乳糖修饰牛奶蛋白的分析。

The food allergies were investigated in 1,514 children at nursery schools in Nagoya city and others. The percentage of diseases was 10%, and 40.6% of patients got sick before the age of one. Egg and milk products were the main allergens for them. The kinds of weaning diets and lactation milks taken were not different among the patient children. In the families with allergic children more members of the families were predispositioned to suffer allergies. Mothers whose children suffered allergies were very fond of foods in their pregnancy and lactation periods.In the present study, we used the monoclonal antibody for specific detection of lactose-protein Maillard adducts as potential milk allergens in commercial milk products such as skim milks, infant powdered milks, pasteurized milks and milk beverages. Meilard adducts were detected with high sensitivity in all eight infant milk samples tested but not in raw milk by ELISA. Furthermore, protein blotting analysis demonstrated that high molecular weight milk proteins such as lactoferrin, BSA and IgG as well as caseins in the infant milks were stained strongly with the monoclonal antibosy. Lactose-protein Maillard adducts were also detected immunochemically in 6 skim milk samples. In the case of skim milks, the high reactivity of monoclonal antibody was observed in lower molecular milk proteins such as beta-LG and alpha-LA as well as caseins. In the case of pasteurized milks, Maillard adducts were hardly detected in LLT milks.Thus this monoclonal antibody is useful for the specific detection of lactose-protein amino carbonyl adducts and the analysis of lactose-modified milk proteins in the complex mixture systems of foods and food products.