白花鬼针草花粉过敏原的分离鉴定与纯化  

Isolation,Identification and Purification of Allergens in Bidens Alba Pollen

作　　者：李莹[1] 肖小军[2] 柴文戍[1] 何韶衡[1] 杨平常[2] 刘志刚[2] 

机构地区：[1]辽宁医学院附属第一医院呼吸内科,辽宁锦州121000 [2]深圳大学医学院过敏反应与免疫学研究所,广东深圳518060

出　　处：《南昌大学学报（医学版）》2014年第6期1-4,共4页Journal of Nanchang University：Medical Sciences

基　　金：国家自然科学基金(811160129;81302553);广东省高等学校国际暨港澳台科技合作创新平台项目(2012gjhz0009);深圳市科技计划国际科技合作项目(GJHZ20130408174112021)

摘　　要：目的提取和分离纯化白花鬼针草花粉主要过敏原,并鉴定主要过敏原的致敏性。方法提取白花鬼针草花粉蛋白粗提液,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分离粗提液蛋白组分并测定其分子质量,经离子交换层析法分离纯化几类主要的蛋白,Western blotting分别检测其与花粉过敏患者血清IgE结合情况及10位正常人阴性混合血清结合情况,对比鉴定每种主要蛋白质致敏原的致敏性。结果 SDS-PAGE结果示：在12.5-120ku之间广泛分布了20余条蛋白条带,其中主带7条,分子质量在35-70ku和10-15ku的区带蛋白含量最为丰富,余下的范围内还有约10余条次带。Westen-blotting检测示：白花鬼针草花粉变应原的致敏性强,与花粉过敏患者血清特异性IgE结合大。离子交换层析出6类主要变应原蛋白,其分子质量分别为70、65、54、49、39及33ku;阴性对照组Westen-blotting在70ku处有弱显色。结论白花鬼针草花粉的主要过敏原为70、65、54、49、39及33ku;70ku变应原致敏性最强。Objective To isolate and purify the allergens from Bidens alba pollen,and to identify the allergenicity of major allergens.Methods Pollen extracts from Bidens alba were prepared and analyzed by SDS-PAGE.The antigens were purified by ion-exchange chromatography.The binding of allergens to serum IgE was detected and the allergenicity of major allergens was identified in patients with pollen allergy and 10normal controls.Results SDS-PAGE analysis showed more than 20protein bands at 12.5-120ku,including 7major bands.The 35-70ku and 10-15ku proteins were the most abundant proteins.Western blotting showed a strong allergenicity of Bidens alba pollen allergens with a extraordinary binding to serum IgE in patients with pollen allergy.Ion-exchange chromatography indicated that the 70,65,54,49,39and 33kDu proteins were the major allergens.In normal controls,Western blotting showed a weak immunoreaction at 70kDu.ConclusionThe 70,65,54,49,39and 33ku proteins are the major allergens and the 70ku protein is the strongest allergen in Bidens alba pollen.

关 键 词：白花鬼针草 花粉变应原 离子交换层系 

分 类 号：R392.8[医药卫生—免疫学]