粉尘螨Der f2变应原的分离纯化及其特征

Purification and Characterization of Der f2 Allergen in Dermatophagoides farinae

目的 用粉尘螨 (Dermatophagoidesfarinae)代谢培养基浸液 (Dff)分离纯化粉尘螨 2类变应原 (Derf2 )。 方法 采用SephadexG 10 0层析、DEAE离子交换层析、PAGE等方法分离纯化Derf2 ,并对Derf2作SDS PAGE测定和热稳定性测定。结果 从Dff分离纯化得到较纯的Derf2 ,经SDS PAGE测定其相对分子质量为 140 0 0 ,并经 10 0℃加热 15min后 ,仍保留 5 0 %～ 83.3 %生物学活性。结论 从Dff中分离纯化的变应原 。

Purpose: To purify Der f2 allergen from the spent culture medium extract of Dermatophagoides farinae mite (Dff). Methods: We purified Der f2 with the aid of Sephadex G-100 gel filtration, DEAE-ion exchange chromatography and polyacrylamide gel electrophoresis, detected the molecular weight of Der f2 using SDS-PAGE method, and analysed the thermostability of Der f2 by heating at 100 °C for 15 min. Results: The pure Der f2 was obtained from Dff. The molecular weight of Der f2 was 14 000. The heated-treated Der f2 retained 50% ∼ 83.3% of the activity of the native allergen. Conclusions: The properties of purified allergen coincided with the characterization of Der f2.