中国食品学报2017，Vol.17Issue(12)：172-181,10.DOI:10.16429/j.1009-7848.2017.12.022
中国对虾蛋白致敏小鼠模型构建及主要过敏原的双向电泳联合质谱鉴定
Establishment of Shrimp (Fenneropenaeus chinensis) Protein-Sensitized Mouse Model and Identification of Major Allergens by 2-D Electrophoresis Combined Mass Spectrometry
摘要
Abstract
Objective:The aims of this work were to establish an allergic model in mice by shrimp protein sensitization and identify the najor allergens in shrimp,Fenneropenaeus chinensis.Methods:The protein was extracted by liquid nitrogen grinding in a lysis buffer.Intraperitoneally injection of protein extracts was used to stimulate allergic responses in ICR mice.The indirect ELISA was employed to test the levels changes of specific IgE and IgG1 antibodies in sera of mice,and the symptom scores were evaluated during different stages of the challenging period.Furthermore,shrimp major allergens were identified by two-dimensional immunoblotting using sera from subjects with shrimp allergy,followed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the peptide digest.Results:The allergic symptoms showed in a dose-dependent manner in mice,and injection of high dose (1.0 mg) of protein can significantly (P＜0.05) increase the levels of specific IgE and IgGl in sera and contribute to the obvious symptoms compared with the control.In addition,two kinds of allergens were identified as arginine kinase (MW:40.2 ku,PI:6.0 and 6.2) and tropomyosin (MW:35.8 ku,PI:4.6) in the species of Chinese shrimp by MALDI-TOF/TOF-MS.Conclusion:Injection of Chinese shrimp proteins is able to evoke the allergic responses in mice.The major allergens are tropomyosin and arginine kinase,and the latter might have two forms of post-translational modifications.关键词
过敏原/双向电泳/免疫印迹/质谱/中国对虾Key words
allergen/two-dimensional electrophoresis/western blotting/mass spectrometry/Chinese shrimp (Fenneropenaeus chinensis引用本文复制引用
傅玲琳,梁亚云,赵淑淑,王彦波..中国对虾蛋白致敏小鼠模型构建及主要过敏原的双向电泳联合质谱鉴定[J].中国食品学报,2017,17(12):172-181,10.基金项目
国家自然科学基金项目(31571770) （31571770）
浙江省自然科学基金项目(LY14C200001) （LY14C200001）
