东亚飞蝗主要过敏原的分析、鉴定与纯化

Purification and identification of allergens in Locusta migratoria manilensis

通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离东亚飞蝗Locusta migratoria manilensis(Meyen)的蛋白质组分并测定其分子量,收集过敏病人血清,采用免疫印迹(Western—blotting)法鉴定其过敏原成分,通过凝胶过滤层析对东亚飞蝗过敏原进行分离纯化。结果表明:东亚飞蝗蛋白粗提液条带大概有30条左右,其中主带大约有10条,相对分子量约为13、15、25、28、40、45、55、70、100、110ku,其中蛋白含量最丰富的约在70ku左右。免疫印迹结果显示,蝗虫过敏条带主要有5条,相对分子量分别约为19、29、38、70、130ku。通过凝胶过滤层析对东亚飞蝗过敏原进行分离纯化,得到了一个高纯度相对分子质量约为70ku东亚飞蝗过敏原,并且发现了一个相对分子质量约为130ku的蝗虫新过敏原。本研究为临床上蝗虫食物变态反应性疾病的诊断和治疗奠定基础。

Proteins from Locusta migratoria manilensis were isolated by SDS—polyacrylamide gel electrophoresis （SDS—PAGE） and identified by Western—blotting using sera from patients allergic to locusts.Allergens from L. m. manilensis were purified by gel filtration chromatography.About 30 protein bands were separated by SDS—PAGE,ten of which were major bands.The proteins, whose molecular weights （MW） were 70 ku, could bind with specific IgE from the sera of locust-allergic patients, thereby identifying them as specific allergens of L. m. manilensis.We identified the primary allergens, whose molecular masses were 19,29,38,70 and 130 ku, by Western-blotting.These results may provide a basis for the diagnosis and immunotherapy of allergic reactions to locusts.