鲤鱼主要变应原的分离、鉴定与纯化

练玉银; 刘志刚; 温岸玲

doi:10.11847/zgggws2006-22-08-34

鲤鱼主要变应原的分离、鉴定与纯化

Purification and identification of major allergens of common carp

摘要

摘要:
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, 16 kDa。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, 36 kDa的变应原组分。
结论鲤鱼主要特异性变应原为42和36 kDa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应原奠定了基础。

Abstract:
Objective To isolate, idntify and purify the major allergens of common carp, which can provide academic basis for clinic.
Methods commoncarp proteins were isolated by SDS-poly acrylamide 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 Chromatag raphy(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 ther apy of fish allergy.

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