梭子蟹过敏原致敏组分的分析研究

Analysis of sensitizing components for sand swimming crab

目的应用免疫印迹(Western blotting)的方法分析梭子蟹[Portunus pelogicus(Linnaeus)]的致敏组分,为蟹过敏原的纯化和标准化变应原疫苗的研制提供理论依据。方法取常规方法制备的梭子蟹浸出液,经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离,用考马斯亮蓝染色分析其抗原蛋白的组分,同时用对蟹过敏的病人血清进行免疫印迹。结果SDS- PAGE显示梭子蟹可辨有19条蛋白带,分子量在13kD-90kD之间,其中主带有9条,分子量是20.9kD、24.2kD、27.1kD、29.2kD、33.7kD、38.9kD、48.7kD、74.7kD、89.1kD。Western blotting结果表明,16例蟹过敏患者血清全部呈阳性反应,浸出液中共有5条致敏条带,其中分子量在74.4kD、48.7kD的是主要致敏组分,阳性反应率均为100%。结论梭子蟹74.4kD和48.7kD组分为主要过敏原。

Objective In order to lay theoretical base for the purification of crab allergens and the standardization of allergenic vaccine, here the antigenic components of sand swimming crab were investigated by Western blot.Methods The crude extract was prepared using routine methods. Separated by SDS-PAGE, antigenic components were detected with Coomassie Brilliant Blue Staining and analyzed by Western blot with sera obtained from crab allergic subjects. Results SDS-PAGE analysis revealed that the proteins of sand swimming crab comprised of at least nineteen discrete protein bands with the molecular weight ranging from 13kD to 90 kD, and the molecular weight of each band is 20.9 kD,24.2 kD, 27.1 kD, 29.2 kD, 33.7 kD, 38.9 kD, 48.7 kD, 74.7 kD, 89.1 kD respectively. Western blot assay showed that five .allergen bands separated from the crude extract were reacting with sera obtained from sixteen crab allergenic subjects,and the bands of 74.4 kD and 48.7 kD were the major allergenic components. The positive rate is 100 %. Conclusions The 74.4 kD and 48.7 kD bands were the major allergens of sand swimming crab.