虾类过敏原的识别、纯化和检测技术研究  

Study on the Identification,Purification and Detection Technique of Shrimp Allergen

作　　者：刘光明[1] 沈苑[1] 曹敏杰[1] 梁银龙[1] 杨景成[1] 苏文金[1] 

机构地区：[1]集美大学生物工程学院福建省高校水产科学技术与食品安全重点实验室,厦门361021

出　　处：《中国食品学报》2008年第6期142-148,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：福建省自然科学基金资助(2006J0419);福建省科技计划重点项目资助(2006I0023;2006F5064)

摘　　要：虾类是人类优质的食用蛋白资源之一,也是联合国粮农组织公布的八大类过敏食物之一。虾类过敏反应严重影响着过敏人群的身体健康和生活质量,为此开展虾类过敏原的识别、纯化和检测技术研究非常必要。通过问卷调查初步了解食物过敏现状,获取自诉虾类过敏患者血清和正常人阴性对照血清,采用特异性IgE检测试剂盒筛选虾类过敏血清。提取南美白对虾蛋白,进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳及免疫印迹识别虾类过敏蛋白,并对患者识别率最高的虾类的主要过敏原进行分离纯化。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、免疫印迹和酶联免疫吸附测定等方法对纯化虾蛋白进行检测分析。患者识别的南美白对虾致敏原的分子质量依次约为200、175、116、85和36kDa,通过硫酸铵盐析及等电点沉淀等方法可以得到电泳纯的虾主要过敏蛋白。免疫印迹结果证实纯化的虾蛋白是具有过敏原性的原肌球蛋白,在此基础上,建立了虾原肌球蛋白的酶联免疫吸附测定方法。Shrimp is one of the best edible protein for human, it is commonly identified as a course of food hypersensitivity, and ingestion of shrimp may course severe allergic reactions such as anaphylactic shock. The potential risk of shrimp allergy was worth to pay more attention. This assay carried out a survey on food allergic status and obtained human serum that had clinical history of shrimp allergy. The allergic and normal sera were selected by specific IgE kit. The results show us that the molecular weights of shrimp allergen recognized by patients are 200 kDa, 175 kDa, 116 kDa, 85 kDa, and 36 kDa. Shrimp tropomyosin was purified to homogeneity by 41%-60% ammonium sulfate fractionation and subjected to isolectric precipitation at pH 4.5 and a protein with molecular weight of 36 kDa was finally obtained. The results were evaluated by means of sodium dodecyl sulfate-polyaerylamide gel electrophoresis（SDS-PAGE） and West- em-blot. On this base, the enzymelinked immunosorbent assays （ELISA） which are used to check and analysis the main allergen of shrimp-tropomyosin were established.

关 键 词：南美白对虾 过敏原 识别 纯化 免疫检测 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程]