油菜花粉过敏原的分析、鉴定与纯化
陈小文 *，尧荣凤，李建春，易金萍
[摘 要] 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。
[中图分类号] R392.8 [文献标识码] A
Analysis, characterization and purification of allergen in oilseed rape pollen
CHEN Xiaowen, YAO Rongfeng, LI Jianqing, YI Jinping
Department of Clinical Laboratory, First Affiliated Hospital of Nanchang University, Nanchang 330006, China
[Abstract] In this paper, we aimed to characterize and purify the allergenic components of oilseed rape
pollen. Firstly, rape pollen coarse extraction fluid was extracted, and the proteins and their molecular were ana－
lyzed by SDS-PAGE. The allergenic components of Oilseed rape pollen extract were separated by ion-exchange
chromatography with DE-52, and then analyzed by Western-blotting. The extract of oilseed rape pollen displayed
10 protein bands, but only four bands (relative molecular weight are 30 000, 25 000, 15 000, and 10 000, re－
spectively) could react with IgE in the sera of patients with oilseed rape pollen allergy. Among the four bands, two
proteins were the major allergen, whose relative molecular weight was 15 000 and 10 000, and found in theⅠ, Ⅱ
and, Ⅲ peaks of ion-exchange chromatography. Our study of preliminary isolation, identification and purification
of the allergenic components in oilseed rape pollen will be used as a base for diagnosis and therapy oilseed rape
pollen related allergy.
[Key words] Oilseed rape pollen; Allergen; Ion-exchange chromatography