鲤鱼主要变应原的分离、鉴定与纯化

目的 对常见的食物过敏原鲤鱼进行特异性抗原成分的分离、鉴定和纯化，为临床诊断及治疗鲤鱼过敏症提供理论依据。方法 通过磷酸盐缓冲溶液进行提取、用十二烷基硫酸钠．聚丙烯胺凝胶电泳（SDS-PAGE）分离出多种蛋白组分并测定其分子量；采用蛋白印迹技术（Western-blotting）鉴定其变应原并通过DE52离子交换层析对鲤鱼变应原进行初步纯化，进一步经凝胶过滤层析对变应原混合组分进行纯化。结果 SDS-PAGE分离出14种蛋白组分，其中主带有9条，分子量分别为5，46，36，34，26，25，23，20，16kDa。Western—blitting结果显示，鲤鱼的主要变应原及次要变应原的分子量是42，36kDa的变应原组分。结论 鲤鱼主要特异性变应原为42和36kDa的蛋白质，为临床诊断和治疗鱼类过敏性疾病提供标准化的廊原尊定了基础．

Purification and identification of major allergens of common carp

Objective To isolate,idntify and purify the major allergens of common carp,which can provide academic basis for clinic.Methods common carp proteins were isolated by SDS-polyacrylamide gel electrophoresie(SDS-PAGE) and the primary and secondary allergen proteins were identified by means of Western-blotting.The major allergens were purified by DEAE-Cellulose DE-52 Ion Exchange Chromatagraphy(IEC),and furthermore one of the primary allergen proteins was purified by Sephadex G-75 Gel Chromatography.It's bioactivity was detected by Western-blotting.Results More than nine major protein bands were obtained by SDS-PAGE(50,46,36,34,26,25,23,20 and 16?kDa),and the primary and secondary allergens were got by Western-blotting(42?kDa 36?kDa).Conclusion The major allergens of common card are 42?kDa,36?kDa,which are most important in establidhing a good quality and standardized fish allergen in the diagnosis and therapy of fish allergy.