紫红笛鲷过敏原的提取、分离及其免疫学特性鉴定

Isolation and Immunological Identification of Allergens in Lutjanus argentimaculatus

目的对紫红笛鲷过敏原进行提取、分离及免疫学特性鉴定。方法新鲜紫红笛鲷经预处理后用PBS缓冲液制备总蛋白粗浸液,SDS-PAGE分析紫红笛鲷总蛋白的组成,免疫印迹(Western-blotting)分析紫红笛鲷过敏原,通过离子交换层析对总蛋白粗浸液进行分离并鉴定不同组份的免疫学特性。结果紫红笛鲷可溶性蛋白粗提液SDS-PAGE显示有20条蛋白条带,对鱼过敏病人的阳性混合血清能与其中7个条带反应,分子量分别是42000,36000,30000,27000,25000,17000和12000Mr。离子交换层析后分子量为42000、36000、12000Mr的阳性过敏原蛋白具有免疫学活性。结论本实验对紫红笛鲷过敏原进行了提取、分离和免疫学特性鉴定,离子交换层析技术可以用于紫红笛鲷过敏原蛋白的分离纯化,为紫红笛鲷过敏原的进一步研究和鱼类食品过敏的防治奠定了理论基础。

Objective To isolate and identify the allergens in red snapper （Lutianus argentimaculatus）, Method Total protein of the red snapper was extracted by PBS and analyzed by SDS-PAGE. The allergens were identified by Western-blotting. To isolate the allergens, ion exchange chromatography was employed. Result Twenty protein bands were observed on the SDS-PAGE gel. The molecular weights of the identified allergens were 42 000, 36 000, 30 000, 27 000, 25 000, 17 000 and 12 000 Mr. Allergens of 42 000, 36 000 and 12 000 Mr were then isolated by ion exchange chromatography. Conclusion The isolated allergens may be used in the future study of allergy to fish proteins.