小清蛋白是鱼类的主要过敏原,三文鱼小清蛋白具有不同的亚型,分子量相近,序列基本相同,给分离纯化带来了不小的挑战.本试验采用硫酸铵盐析、Qxl-Sepharose离子交换结合凝胶过滤等方法定向纯化小清蛋白β1型,并应用免疫印迹法对纯化的小清蛋白进行过敏原性鉴定,结合激光辅助解析/飞行时间质谱进行结构鉴定.结果显示,硫酸铵盐析、离子交换层析结合凝胶过滤纯化方法可以得到纯度达90%以上的小清蛋白β1型,且蛋白具有很强的免疫反应活性.针对该蛋白的研究不仅有利于过敏原检测方法的建立,也可为低致敏性水产品的开发提供理论依据.%Parvalbumin is the major allergen of fish species. The different subtypes of salmon parvalbumin with close molecular weight and sequence bring great challenges to protein separation and purification. Parvalbu-minβ1 was purified by heat treatment,(NH4)2SO4 salting out method and Qxl-Sepharose chromatography com-bined by gel filtration. Western blotting was used to identify the allergenicity of purified Parvalbumin. MALDI-TOF/MS was used to further identify the characteristic of parvalbumin structure. The results revealed that the purity of the parvalbuminβ1 was more than 90 percent and it had immunological reactive activity. Investigation into this protein is beneficial not only to detecting allergen, but also to producing aquatic products with low aller-genicity.
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