基础医学与临床 ›› 2011, Vol. 31 ›› Issue (9): 1036-1039.

• 技术与方法 • 上一篇    下一篇

重阳木花粉过敏原的分析、鉴定与纯化

姚敏   

  1. 中南大学湘雅医学院附属海口市人民医院
  • 收稿日期:2010-05-07 修回日期:2010-07-23 出版日期:2011-09-05 发布日期:2011-09-05
  • 通讯作者: 姚敏 E-mail:yaoruan@sina.com
  • 基金资助:
    国家自然科学基金;省科技发展项目

Analysis, characterization and purification of allergen in Bischofia polycarpa pollen

  • Received:2010-05-07 Revised:2010-07-23 Online:2011-09-05 Published:2011-09-05

摘要: 【摘要】 目的 对重阳木花粉变应原蛋白进行分析、鉴定与纯化。方法 提取这重阳木花粉的粗提液,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS - PAGE)分离粗提液蛋白质组分并测定其分子量,收集过敏病人血清,采用免疫印迹(Western - blotting)法鉴定其变应原成分,通过离子交换层析对重阳木花粉变应原进行初步纯化和免疫印迹鉴定。结果 重阳木花粉有18条主要蛋白带,12 000Mr和14 000Mr为重阳木花粉特异性变应原;通过离子交换层析方法纯化出重阳木花粉分子量为12 000Mr和14 000Mr的变应原主要分布在II峰中。结论 对重阳木花粉变应原进行了初步的分离、鉴定和纯化,为临床重阳木花粉过敏疾病的诊断和治疗奠定了基础。

关键词: 重阳木花粉 , 变应原 , 离子交换层析

Abstract: 【Abstract】 Objective To characterize and purify the allergenic components of Bischofia polycarpa pollen. Methods Pollen extract from Bischofia polycarpa pollen prepared and analyzed by SDS-PAGE and proteins band were visualized by staining the gel with Coomassis blue. Antigenic properties of the separated proteins were analyzed by immuno-blotting. Ion-exchange chromatography with DE-52 was used for the purification of the antigens. Then purified products were characterized by western-blotting. Results For the pollen extract of Bischofia polycarpa, 18 protein bands was in SDS-PAGE. Two of these protein bands, with a molecular weight of 12 000 Mr ang 14 000 Mr, showed immunoreactivity with IgE in the sera from patients with pollen allergy of Bischofia polycarpa. Result from the ion-exchange chromatography indicated that the two pollen allergens (12 000 、14 000 Mr) from Bischofia polycarpa were eluted in the second peak. Conclusion Pollen allergens form Roystonea regia were purified and characterized.The result will provide a basis for the diagnosis and treatment of allergy to the pollen allergens form Bischofia polycarpa

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