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目的分离纯化刀额新对虾的主要变应原蛋白并对其进行免疫学鉴定和应用。方法刀额新对虾蛋白采用DEAE-52离子交换色谱和Sephadex G-100凝胶过滤纯化,各组分用斑点酶免疫法检测,活性组分用虾过敏症患者血清以ELISA方法检测其免疫反应性,分子量用SDS-PAGE测定。结果纯化得到2种刀额新对虾变应原,在ELISA检测中显示良好的免疫反应性,其分子量分别为36kDa和68kDa。结论经离子交换层析和分子筛层析纯化的刀额新对虾主要变应原保持良好的免疫学活性,可作为特异性抗原应用于ELISA诊断试剂的制备。
OBJECTIVE: To isolate and purify the major allergens of Penaeus vannamei and to identify and apply it for immunological identification. Methods The new shrimp protein was purified by DEAE-52 ion exchange chromatography and Sephadex G-100 gel filtration. The components were detected by dot enzyme immunoassay. The active components were detected by ELISA with serum from shrimp allergy sufferers , Molecular weight by SDS-PAGE. Results Two species of shrimp newborn shrimp allergen were purified and showed good immunoreactivity in ELISA. The molecular weights were 36kDa and 68kDa, respectively. Conclusion The main allergens of P. xylostella purified by ion exchange chromatography and molecular sieve chromatography have good immunological activity and can be used as specific antigens in the preparation of ELISA diagnostic reagents.