大黄鱼过敏原的提取、分离及免疫学特性鉴定

Extraction,isolation and immunological identification of allergens from Pseudosciaena crocea

目的提取、分离大黄鱼特异性过敏原成分,并对其免疫学特性进行鉴定。方法采用Coca’s提取液提取大黄鱼蛋白;十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析大黄鱼总蛋白的组成,以对鱼过敏患者阳性血清为一抗,Western-Blotting分析大黄鱼中的过敏原;阴离子交换层析对大黄鱼总蛋白进行初步分离,Western-Blotting分析各个组份的免疫学活性。结果大黄鱼粗浸液蛋白分子量在8～116kD之间;Western-Blotting检测到分子量为54、29、27和14kD的过敏原;通过离子交换层析分离,过敏原蛋白的相对含量有显著提高,且大多都保持原有的免疫学活性。结论研究发现了大黄鱼中多种过敏原并对其免疫学活性进行了鉴定。

Objective To extract,identify and purify the major allergens of Pseudosciaena crocea in order to evaluate the immunological activities. Methods Pseudosciaena crocea proteins were extracted by Coca s buffer. Allergens of the Pseudosciaena crocea were identified by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western-Blotting. Major allergens were isolated by ion exchange chromatagraphy (IEC) and their immunological activities were evaluated by Western-Blotting with the serum of patients as 1st antib...