摘要
采用液氮研磨法提取葎草花粉粗蛋白,凝胶电泳(SDS.PAGE)分离粗提液蛋白组分并测定各组分的相对分子质量,后经过离子交换层析法分离纯化几类主要的蛋白,再用westenblotting分别检测其与花粉过敏患者血清IgE结合情况,鉴定每种主要蛋白质致敏原的致敏性.分析了葎草花粉变应原成分及其变应原性、免疫原性,并鉴定主要过敏原的致敏性.SDS-PAGE凝胶电泳结果表明,广泛的分布于8—170kDa之间有主带13条,次带10余条;westenblotting检测发现蓓草花粉的致敏性较强,与花粉过敏患者血清IgE结合能力较高.离子交换层析出4种主要变应原蛋白,其相对分子质量分别为55,16,15和25kDa,其中结合能力最强的是15kDa处蛋白条带.蓓草的主要过敏原为55,16,15和25kDa蛋白质组分.
To characterize and purify the pollen allergens isolated from Humulus scandens Lour, identified the aller- genicity of these main allergen. The pollen of Humulus scandens Lour were grinded in liquid nitrogen to extract the total protin, analyzed total protin with SDS-PAGE, detective the proteins' allergenicity with westen-blotting, Ion-ex- change chromatography with DE-52 was used for the purification of the antigens. For the pollen extract of Humulus scandens Lour,30 protein bands were detected in SDS-PAGE. Among them,weights bewteen 8 and 170 kDa has 13 main protiens, more than 10 kinds of secondary protiens. Fore of these protein bands, with a molecular weight of 55 kDa, 16 kDa and 15 kDa,25 kDa,showed immunoreactivity with IgE in the sera from patients with allergy to pol- len of Humulus scandens Lour. Pollen allergens from Humulus scandens Lour were purified and characterized. The re- sult will provide a theoretical foundation for the diagnosis and treatment of allergy to the pollen allergens from Hu- mulus scandens Lour.
出处
《江西师范大学学报(自然科学版)》
CAS
北大核心
2014年第1期26-29,共4页
Journal of Jiangxi Normal University(Natural Science Edition)
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(GJHZ20130408174112021)资助项目
关键词
葎草
花粉变应原
离子交换层析
Humulus scandens Lour
pollen allergen
ion-exchange chromatography