摘要
目的:以南美白对虾中主要过敏原原肌球蛋白(TM)为研究对象,研究TM在体外模拟胃液(SGF)消化中表位的变化情况。方法:SDS-PAGE、Western blot和间接ELISA用于分析TM在SGF中的消化情况以及免疫活性和过敏原性的变化,并采用质谱方法分析TM抗原表位的变化。结果:SDS-PAGE和Western blot结果显示,随着消化时间的增加,TM条带逐渐变浅,但是在2 h时仍具有免疫活性。间接ELISA结果显示,消化2 h后,TM的免疫活性及过敏原性分别下降89.1%和80.8%。质谱结果表明,TM在胃液消化过程中大部分抗原表位会被破坏,但是TM在胃液中消化2 h时仍具有免疫活性和过敏原性,很可能是因为消化片段中存在有3个不被破坏的抗原表位(肽段111~125、137~141和153~161)和4个抗消化能力很强的抗原表位(肽段50~66、144~151、145~164和150~163)所致。结论:本研究可为今后探讨人体消化对TM过敏原性的影响机理及脱敏或低致敏性食品的开发提供科学依据。
Objective:Taking the main allergen tropomyosin(TM)in Litopenaeus Vannamei as the research object,to study the epitopes changes of TM in vitro simulated gastric fluid(SGF)digestion.Methods:The digestion of TM and the changes of immunological activity and allergenicity in SGF were analyzed by SDS-PAGE,Western blot and indirect ELISA methods,and the epitopes changes of TM were analyzed by mass spectrometry.Results:SDS-PAGE and Western blot results showed that the TM band became lighter with the increase of digestion time,but it was still immunologically active at 2 h.The results of indircet ELISA showed that after 2 h digestion,the immunological activity and allergenicity of TM lost 89.1%and 80.8%,respectively.Mass spectrometry results showed that most of the epitopes of TM were destroyed during SGF.However,TM was still immunologically active and allergenic after it was digested in SGF for 2 hours,probably because there were three undestroyed epitopes(peptides 111-125,137-141 and 153-161)and four antigenic epitopes that were highly resistant to digestion(peptides 50-66,144-151,145-164 and 150-163)in digestion fragments.Conclusion:This study can provide the scientific basis for exploring the mechanism of human digestion on the allergenicity of TM and the development of desensitized or hypoallergenic foods.
作者
王学丽
李娅茹
王梦梦
丁婕
卢瑛
WANG Xue-Li;LI Ya-Ru;WANG Meng-Meng;DING Jie;LU Ying(College of Food Science and Technology,Shanghai Ocean University,Shanghai Engineering Research Center of Aquatic Product Processing&Preservation,Laboratory of Quality&Safety Risk Assessment for Aquatic Products on Storage and Preservation(Shanghai),Ministry of Agriculture,Shanghai 201306,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第19期2319-2325,共7页
Chinese Journal of Immunology
基金
上海市科技兴农重点攻关项目[沪农科攻字(2016)第4-4号]
上海市科学技术委员会工程研究中心推广项目(16DZ2280300)
