摘要
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。
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.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2015年第10期1375-1379,1388,共6页
Chinese Journal of Immunology
基金
国家自然科学基金面上项目课题(No.81371882)
关键词
中华绒鳌蟹
特异性s
Ig
E
血蓝蛋白
质谱分析
过敏原
特异性sIgE
Eriocheir sinensis
Specific sIgE
Hemocyanin
MALDI-TOF/TOF analysis
Allergen