田菁花粉过敏原的分离、纯化及鉴定

Isolation,Purification and Identification of Allergens in Sesbania Pollen

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27ku为其特异性的过敏原,其中16和19ku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19ku。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 protein bands.The immunoreactive bands at 16,19 and 27 ku were specific allergens and the immunoreactive 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 Sesbania pollen provide a basis for the diagnosis and treatment of allergic diseases.