不同加工方式降低大黄鱼鱼卵过敏原及其消化产物免疫反应性的比较研究

Comparative research on different processing methods decreasing the immunoreactivity of the roe of large yellow croaker allergen and digestion products

目的本文以大黄鱼鱼卵为原料,分析比较不同加工方式对大黄鱼鱼卵过敏原及其消化产物免疫反应性的影响。方法采用不同加工方式(美拉德反应、紫外照射、超声联合加热、高温高压)处理鱼卵,制备卵黄蛋白粗提物并进行模拟胃肠液二相消化实验,利用免疫印迹及抑制性ELISA方法对粗提物及消化产物中过敏原的IgG/IgE结合活性进行分析。结果分析不同加工方式处理后卵黄蛋白中过敏原的IgG结合活性,结果发现当抗体稀释倍数为4×104时,检测不到美拉德反应产物与抗体发生特异性结合,而经后三种物理加工方式处理后的粗提物均与抗体有不同程度的免疫反应。此外,分析加工处理及其消化产物中过敏原与患者血清的IgE结合活性,结果显示经不同加工方式处理后,仅有美拉德反应产物的抑制率低于50%;经高温高压处理的粗提物抑制率为65%,但消化后其抑制率降至20%以下。结论在4种不同的加工方式中,美拉德反应能有效地降低大黄鱼鱼卵中过敏原的IgG/IgE结合活性,超声联合加热及高温高压处理明显减弱过敏原消化产物的IgE结合活性,紫外照射对过敏原的影响不明显。

Objective To analyze and compare the effects of different processing methods on the immu-noreactivity of the roe of large yellow croaker allergen and digestion products. Methods The crude extracts of yolk protein from large yellow croaker were performed to simulate gastric and intestinal fluid two phase digesti on after extracted by 4 different processing methods (Maillard reaction, ultraviolet radiation, ultrasonic-heating, high pressure steaming). Then the IgG/IgE-binding ability of the allergen in crude extracts and digestion prod-ucts was analyzed by Western-blotting and inhibition ELISA methods. Results The IgG-binding ability of the allergen in yolk protein was analyzed after treated by different processing methods. The results showed that it could not be detected the specific binding between Maillard reaction products and antibody, when the antibody was diluted 4×104 times. However, the crude extracts could react with antibody inordinately after treated by 3 physical processing methods. In addition, analyzing the IgE-binding ability of the processed and digested al-lergen with patients’ sera found that only the inhibition ratio of Maillard reaction products was lower than 50%after treated by different processing methods, but its inhibition ratio varied inconspicuously. Nevertheless, the inhibition ratio of the crude extracts treated by high pressure steaming was 65%, while its inhibition ratio dropped to below 20% after digestion. Conclusion In the different processing methods, Maillard reaction could effectively decrease the IgG/IgE-binding ability of the allergen in the roe of large yellow croaker. Ultra-sonic-heating and high pressure steaming could decrease the IgE-binding ability of its digestion products ob-viously, although the IgG/IgE-binding ability of the allergen in large yellow croaker was not decreased. But ul-traviolet radiation had little effect on the immunoreactivity of the allergen.