血蓝蛋白是中华绒鳌蟹重要的高分子量过敏原

摘要

Objective:To prove hemocyanin is an important high-molecular-weight allergen from Eriocheir sinensis.Methods:The proteins were extracted from Eriocheir sinensis tissue with lysate extraction method .The protein components were analyzed by SDS-PAGE,two-dimensional electrophoresis and Western blot.Serum IgE from allergic donors identified a candidate protein,which was char-acterized by MALDI-TOF/TOF.The immune property of the candidate protein was tested using Dot-blot.Results: By SDS-PAGE and two-dimensional electrophoresis analysis,we prove that the native protein components were completely.According to the Western blot result,at least 18 components could react with the positive serum.Among them, two 70-80 kD proteins had the same isoelectric point.So,may be they were the same protein.MALDI-TOF/TOF analysis results showed that the suspected proteins were hemocyanin.A sensitization frequency of 61%was observed in Eriocheir sinensis patients by Dot-blot.Conclusion:Hemocyanin was identified as an important high-molecular-weight allergen from Eriocheir sinensis.%目的：证明血蓝蛋白是中华绒螯蟹的一种重要的高分子量过敏原。方法：采用裂解液法提取蟹总蛋白，通过SDS－PAGE、二维电泳及Western blot分析蟹中过敏原组分并进行比对，再通过质谱分析鉴定未知过敏原。以临床中华绒螯蟹过敏患者的血清作为识别抗体，通过斑点免疫印记实验证明疑似蛋白的致敏特性。结果：通过SDS－PAGE及二维电泳分析，证明本实验所提取的蟹总蛋白组分完全。 Western blot结果显示，中华绒螯蟹至少有18种与阳性血清反应的组分，其中，70～80 kD处两个蛋白处于同一等电点，可能为同一蛋白，且反应斑点清晰；质谱分析结果显示疑似蛋白为血蓝蛋白；斑点免疫印记结果显示血蓝蛋白与患者血清的反应频率为61％。结论：血蓝蛋白为中华绒鳌蟹中一种重要的高分子量过敏原。