几种常见食物过敏原的纯化及免疫活性鉴定

发布时间：2018-10-12 18:33

【摘要】：[目的] 通过建立食物过敏模型,确立并优化sIgE和组胺定向释放的检测方法,为临床提供可替代皮内试验的持异、灵敏、安全的多过敏原体外诊断过敏症技术,并为重组表达过敏原,实现过敏原的标准化作前期研究。 [方法] 制备牛奶、蟹和鳖等几种常见的食物过敏原,建立食物过敏症动物模型,间接ELISA法检测抗血清中特异性IgE和IgG水平,荧光法测定致敏腹腔肥大细胞的组胺定向释放,免疫印迹分析过敏原的过敏组分,饱和硫酸铵分段赫析法分离纯化过敏组分。 [结果] 用乳清蛋白和蟹蛋白免疫BALB/c小鼠成功建立了食物过敏模型,模型组小鼠血清sIgE和组胺水平均比对照组明显升高。用氢氧化铝作佐剂,以皮下注射途径免疫小鼠获得了理想的免疫效果。连续两代用不含牛奶或奶制品的特制饲料饲养的小鼠sIgE水平低,显示了特制饲料对过敏模型构建的重要性。激发后第14d的定向组胺释放率显示组胺的测定与sIgE检测结果具有良好的一致性。 [结论] 腹腔肥大细胞体外定向组胺释放率和sIgE水平检测可用于食物过敏原鉴定与评价,对研究食物过敏症的诊断和治疗具有十分重要的意义,亦可用于转基因食物蛋白过敏原的检测。
[Abstract]:[objective] to establish and optimize the detection method of sIgE and histamine directional release by establishing food allergy model, and to provide a reliable, sensitive and safe technique for the in vitro diagnosis of allergy with multiple allergens, which can be used as a substitute for intradermal test. And for the recombination of the expression of allergens and achieve the standardization of allergens for the preliminary study. [methods] several common food allergens such as milk, crab and turtle were prepared, and animal models of food hypersensitivity were established. Specific IgE and IgG levels in antiserum were detected by indirect ELISA assay. Histamine release from sensitized peritoneal mast cells was determined by fluorescence method. The allergy components of the allergen were analyzed by Western blotting, and the anaphylaxis was isolated and purified by saturated ammonium sulfate. [results] BALB/c mice were immunized with whey protein and crab protein. The serum sIgE and histamine levels in the model group were significantly higher than those in the control group. Using aluminum hydroxide as adjuvant, the mice were immunized by subcutaneous injection. The low sIgE level of mice fed with special feed without milk or dairy products for two consecutive generations showed the importance of special feed for the construction of allergic model. The release rate of directed histamine on the 14th day after excitation showed a good agreement with the results of sIgE. [conclusion] the detection of specific histamine release rate and sIgE level of peritoneal mast cells in vitro can be used to identify and evaluate food allergens, and it is of great significance to study the diagnosis and treatment of food allergies. It can also be used for the detection of protein allergens in genetically modified foods.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392.1

【相似文献】