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| 粉尘螨变应原Derf1基因的原核表达及多克隆抗体的制备 | |
| 引用本文： | 石连,姜玉新,王海宁,李朝品.粉尘螨变应原Derf1基因的原核表达及多克隆抗体的制备[J].皖南医学院学报,2012,31(1):3-6. |
| 作者姓名： | 石连 姜玉新 王海宁 李朝品 |
| 作者单位： | 1. 安徽理工大学医学院 病原生物学与免疫学教研室,安徽淮南,232001 2. 皖南医学院 医学寄生虫学教研室,安徽 3. 安徽理工大学医学院 病原生物学与免疫学教研室,安徽淮南232001;皖南医学院 医学寄生虫学教研室,安徽芜湖241002 |
| 基金项目： | 国家自然科学基金项目(30872367);安徽省自然科学基金项目(070413088) |
| 摘 要： | 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pET-28a(+)-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。 |
| 关 键 词： | 粉尘螨 变应原 Derf1 多克隆抗体 |
Prokaryotic expression and preparation of polyclonal antibody of recombinant group 1 allergen from Dermatophagoides farinae |
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| SHI Lian , JIANG Yu-xin , WANG Hai-ning , LI Chao-pin.Prokaryotic expression and preparation of polyclonal antibody of recombinant group 1 allergen from Dermatophagoides farinae[J].Acta Academiae Medicinae Wannan,2012,31(1):3-6. | |
| Authors: | SHI Lian JIANG Yu-xin WANG Hai-ning LI Chao-pin |
| Institution: | School of Medicine,Anhui University of Science and Technology,Huainan 232001,China |
| Abstract: | Objective:To purify the recombinant protein of Dermatophagoides farinae(Der f1) through prokaryotic expression in Escherichia coli and prepare its polyclonal antibody.Methods: After recombinant plasmid Der f1/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 electrophoresis and purified.Anti-Der f1 antibody was raised by immunization against Der f1 in New Zealand white rabbits through detection of specificity using Western blot method.Results: Def f1 was recovered by gel extraction,and the titer of anti-Der f1 antibody was 1∶32 000.Conclusion: Anti-Der f1 antibody was prepared successfully,which will provide a base for diagnosis and immunotherapy of allergic diseases. |
| Keywords: | Dermatophagoides farinae allergen Der f1 polyclonal antibody |
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