Objective-.To purify the recombinant protein of Dermatopha-goides farinae(Der fl) through prokaryotic expression in Escherichia coli and prepare its polyclonal antibody. Methods-. After recombinant plasmid Der/l/pET-28a( + )Escherichia coli BL21 was successfully constructed, the bacterial pellet was resuspended and lysed by freezing-thawing treatment. The target protein was excised from the gel after SDS-PAGE electro-phoresis and purified. Anti-fler fl antibody was raised by immunization against Derfl in New Zealand white rabbits through detection of specificity using Western blot method. Results; Def fl was recovered by gel extraction,and the titer of mli-Derfl antibody was 1:32 000. Conclusion-. Anti-Der fl antibody was prepared successfully,which will provide a base for diagnosis and immunotherapy of allergic diseases.%目的:原核表达、纯化粉尘螨主要变应原Der f1重组蛋白,并制备其多克隆抗体.方法:大量诱导表达含pET28a(+)-Der f1质粒的BL21菌株,SDS-PAGE电泳并割胶纯化目的蛋白,以此为抗原免疫新西兰大白兔,收集血清进行效价检测并用Western blot检测其特异性.结果:割胶回收可有效纯化Der f1重组蛋白,抗Der f1抗体效价为1∶32 000.结论:成功制备Der f1多克隆抗体,以期为尘螨过敏性疾病的诊断和免疫治疗提供参考.
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