屋尘螨主要过敏原蛋白Der p1的分离纯化

Isolation and purification of Der p1,major allergen of Dermatophagoides pteronyssinus

目的分离纯化出屋尘螨中主要过敏原蛋白Der p1。方法使用PBS-T溶液对屋尘螨干粉中的过敏原蛋白进行粗提取,利用阳离子交换层析对经过透析后粗提取液中的Der p1进行分离纯化,采用30%的NaCl溶液(1.0 mol/L)进行洗脱。然后通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)对透析后洗脱液中的蛋白质进行鉴定。结果屋尘螨中有多条蛋白区带,其中相对分子质量25000为主要过敏原蛋白,屋尘螨通过阳离子交换层析纯化出相对分子质量25000的蛋白组,主要集中在Ⅲ峰。结论初步分离纯化及鉴定了屋尘螨过敏原蛋白Der p1,为临床过敏性疾病的诊断和治疗奠定了基础。

Objective The aim of this study was to isolate and purify Der p 1,a major allergen of Dermatophagoides pteronyssinus.Methods The allergen proteins were extracted crudely from dry powders of Dermatophagoides pteronyssinus by using PBS-T solution.Der p1 in crude extract of allergen proteins was isolated and purified by cation exchange chromatography,eluting with 30%NaCl(1.0 mol/L)after dialysis.The purified proteins in eluants after dialysis were identified by using sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE).Results Many protein bands were demonstrated in extract of Dermatophagoides pteronyssinus.The 25000 protein was identified as the major allergen.The result of cation exchange chromatography indicated that the 25000 was eluted and collected in PeakⅢ.Conclusion er p1,the major allergen of Dermatophagoides pteronyssinus was preliminarily isolated,purified and identified,which provide a basis for clinical diagnosis and treatment of allergic diseases.