Ma Xiaojuan, Liang Rui, Yang Xiaotong, Gou Jingkun, Li Yan, Lozano-Ojalvo Daniel
School of Public Health, Zunyi Medical University, Zunyi 563000, China.
School of Public Health, Zunyi Medical University, Zunyi 563000, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2020 Sep 1;1152:122231. doi: 10.1016/j.jchromb.2020.122231. Epub 2020 Jun 10.
Hen egg is a worldwide top-consumed food that has attracted public health concerns because it can induce allergic reactions in sensitized individuals. Food allergy investigations need highly purified egg allergens. However, a limited number of purification methods have been described for the combined separation of more than two egg allergens and only few of them have evaluated the immunological activity of these purified proteins. The aim of this work was to develop a chromatographic method for the separation of the four major egg allergens (ovomucoid, ovalbumin, ovotranferrin, and lysozyme) with a demonstrated immunological activity. After a pre-processing step for ovomucin precipitation and pH adjustment, remaining egg white proteins were loaded onto CM-Sepharose column and major egg allergens were separated using cation-exchange chromatography. Yield of ovomucoid, ovalbumin, ovotranferrin, and lysozyme was 60.0%, 52.1%, 29.6%, and 90.2%, respectively. Purified allergens were compared with their commercial standards, showing a high purity as well as a maintained antigenicity. The protocol described in this work is simple, quick, low-cost, and suitable for the study of the immunological properties of these allergens. For higher ovalbumin demand in the lab, 2.1 g ovalbumin can be produced in a single process with high purity.
鸡蛋是一种全球消费量很大的食物，由于它会在过敏个体中引发过敏反应，因此引起了公众对健康问题的关注。食物过敏研究需要高度纯化的鸡蛋过敏原。然而，针对两种以上鸡蛋过敏原的联合分离所描述的纯化方法数量有限，并且其中只有少数方法评估了这些纯化蛋白的免疫活性。这项工作的目的是开发一种色谱方法，用于分离具有已证实免疫活性的四种主要鸡蛋过敏原（卵类粘蛋白、卵清蛋白、卵转铁蛋白和溶菌酶）。在进行卵粘蛋白沉淀和pH调节的预处理步骤后，将剩余的蛋清蛋白加载到CM-琼脂糖柱上，并使用阳离子交换色谱法分离主要的鸡蛋过敏原。卵类粘蛋白、卵清蛋白、卵转铁蛋白和溶菌酶的产率分别为60.0%、52.1%、29.6%和90.2%。将纯化的过敏原与其商业标准品进行比较，结果显示其具有高纯度以及保持的抗原性。这项工作中描述的方案简单、快速、低成本，适用于研究这些过敏原的免疫学特性。对于实验室中对卵清蛋白的较高需求，单次生产过程中可生产出2.1克高纯度的卵清蛋白。
