利用牵出技术分析与虾主要过敏原相互作用的蛋白质

Applying Pull-down Technique to Study Interactional Proteins with Shrimp Allergen

目的:应用牵出技术研究与虾主要过敏原相互作用的蛋白质。方法:将纯化的虾主要过敏原原肌球蛋白偶联在用环氧氯丙烷活化的载体Sepharose-4B上作为配基并制成层析柱,以过敏原为饵蛋白从虾过敏患者血清中钓出与之相互作用的蛋白质。结果:SDS-PAGE和间接ELISA检测显示,由过敏原牵出的蛋白质至少有两种,分别是抗原特异性IgE和IgG,这说明虾过敏反应除了是由IgE所介导外,IgG也在其中起到一定的作用。结论:实验所采用的牵出技术是一个快速且有效的研究蛋白质相互作用的方法,其基于免疫亲和层析原理。

Objective： The interactional proteins with shrimp main allergens were studied by pull-down technology. Methods ： Sepharose 4B polymer beads were activated by using epichlorohydrin, shrimp main allergen （tropomyosin） was coupled to them to prepare a chromatography column, which was applied to capture the interactional proteins with shrimp main allergens from patient＇ s serum. Results： SDS-PAGE and indirect ELISA showed that at least two proteins in patient＇ s serum were captured by shrimp allergen, they were aUergen-specific IgE and IgG respectively. The result showed that the allergy is mediated not only by IgE, IgG also plays a role. Conclusion： The pull-down technique performed is a rapid and effective method to study protein-protein interactions and is based on the immune affinity chromatography principle.