目的：初步分离、纯化及鉴定田菁花粉变应原蛋白。方法对田菁花粉粗提液进行提取，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）对粗提液蛋白组分进行分离，并对其分子质量进行测定。收集10例过敏患者血清，通过免疫印迹法（Western-blotting）对花粉变应原成分进行鉴定，用离子交换层析对其变应原进行初步纯化，并通过免疫印迹法进行鉴定。结果田菁花粉有蛋白条带20余条，其中主要条带有12条，16、19和27 ku为其特异性的过敏原，其中16和19 ku 为其主要过敏原。田菁花粉通过离子交换层析纯化出变应原主要集中在Ⅳ和Ⅰ峰，其对应的分子质量为16和19 ku。结论初步分离、纯化及鉴定了田菁花粉变应原，为临床过敏性疾病的诊断和治疗奠定了基础。%Objective To tentatively isolate,purify and identify the allergens in Sesbania pol-len.Methods Proteins were isolated from Sesbania pollen extracts by SDS-PAGE and molecular weight was determined.Serum samples were collected from 10 Sesbania pollen allergic patients and antigenic properties of Sesbania pollen were analyzed by Western-blotting.The antigens were purified by ion-exchange chromatography and the purified products were identified by Western-blotting.Results About 20 protein bands were observed,and 12 of them were typical major pro-tein bands.The immunoreactive bands at 16,19 and 27 ku were specific allergens and the immu-noreactive bands at 16 and 19 were main allergens.The ion-exchange chromatography indicated that the two allergens (16 ku and 19 ku)from Sesbania pollen were eluted in the fourth and first peaks,respectively.Conclusion The isolation,purification and identification of allergens in Ses-bania pollen provide a basis for the diagnosis and treatment of allergic diseases.
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