论文部分内容阅读
目的对油菜花粉的变应原组分进行鉴定及初步的分离及纯化。方法提取油菜花粉粗提液,然后通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离油菜花粉的蛋白质组分并测定其相对分子质量,采用免疫印迹(Western blotting)法鉴定其变应原成分,并通过离子交换层析对油菜花粉变应原进行初步分离纯化,免疫印迹进行检测。结果油菜花粉粗提液有10余条蛋白带,其中相对分子质量为30 000、25 000、15 000和10 000的蛋白可与油菜花粉过敏性病人血清IgE结合,其中15 000和10 000为主要变应原;离子交换层析结果显示主要过敏原成分主要分布在Ⅰ、Ⅱ和Ⅲ峰中。结论对油菜花粉变应原进行了初步的分离、鉴定和纯化,为临床油菜花粉变态反应疾病的诊断和治疗奠定了基础。
Objective To identify and preliminary isolation and purification of allergen components of rape pollen. Methods The crude extract of rape pollen was extracted. The protein components of rape pollen were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the relative molecular mass was determined. Western blotting Identification of allergen components, and by ion exchange chromatography rape pollen allergen preliminary separation and purification, Western blot detection. Results There were more than 10 protein bands in the rape pollen crude extract. The proteins with relative molecular mass of 30 000, 25 000, 15 000 and 10 000 could bind with IgE in the rape pollen allergic patients, 15 000 and 10 000 Allergen; ion exchange chromatography showed that the main allergen components were mainly distributed in Ⅰ, Ⅱ and Ⅲ peaks. Conclusion Preliminary isolation, identification and purification of rape pollen allergens laid the foundation for the diagnosis and treatment of clinical rape pollen allergy diseases.