双抗体夹心酶联免疫吸附法检测杏仁过敏原苦杏仁球蛋白  

Development of Double-Antibody Sandwich ELISA for the Detection of the Almond Allergen Amandin

作　　者：张洁琼[1] 高淑霞[1] 生威[1] 张燕[1] 王硕[1] 

机构地区：[1]天津科技大学食品工程与生物技术学院,教育部食品营养与安全重点实验室,天津300457

出　　处：《食品科学》2013年第16期173-177,共5页Food Science

基　　金："十二五"国家科技支撑计划项目(2011BAK10B03);天津市科技计划项目(10SYSYJC28300)

摘　　要：提取中国甜杏仁过敏原蛋白苦杏仁球蛋白,分别制备兔和鼠多克隆抗体,建立双抗体夹心酶联免疫吸附(ELISA)法。结果表明:对甜杏仁中苦杏仁球蛋白的检测限为(6.36±1.02)μg/L;对中国苦杏仁及美国大杏仁中苦杏仁球蛋白的检测限分别为(12.11±1.70)μg/L和(18.95±1.52)μg/L,且与常见的14种植物性蛋白没有交叉反应,表明该方法具有良好的特异性。将该方法用于法式小面包、饼干和脱脂牛乳中杏仁过敏原的检测,苦杏仁球蛋白的添加回收率为78.94%～125.15%,且相对标准偏差均低于5.81%。In the present study,allergen amandin from Chinese sweet almond was prepared,and specific antibodies against the amandin were produced.A double-antibody sandwich enzyme linked immunosorbent assay（ELISA） based on rabbit polyclonal antibody and mouse polyclonal antibody was then established.The newly established ELISA could detect amandin at levels as low as（6.36 ± 1.02） μg/L,showing no cross-reactivity with 14 common plant proteins.The limit of detection for amandin was（12.11 ± 1.70） μg/L in Chinese bitter apricot kernel and（18.95 ± 1.52） μg/L in almond（AmygdaluscommunisL.）.The sandwich ELISA were used to detect the amandin in spiked samples of Frenchstyle bread,biscuit and skim milk with recoveries ranging from 78.94% to 125.15%,and relative standard deviation（n = 3） below 5.81%.

关 键 词：杏仁 苦杏仁球蛋白 双抗体夹心酶联免疫吸附分析法 

分 类 号：R155.5[医药卫生—营养与食品卫生学]