双抗体夹心ELISA检检测仁过敏原Amandin  

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

作　　者：李明新 巴巨伟 刘俊伟 陆旸[1] LI Mingxin;BA Juwei;LIU Junwei;LU Yang(College of Food Science and Engineering,Tianjin University of Science and Technology,Tianjin 300457,China;Tianjin Binhai New Area Urban and Rural Residents Basic Medical Insurance Service Center,Tianjin 300456,China)

机构地区：[1]天津科技大学食品科学与工程学院,天津300457 [2]天津市滨海新区城乡居民基本医疗保险服务中心,天津300456

出　　处：《天津科技大学学报》2025年第2期44-50,共7页Journal of Tianjin University of Science & Technology

摘　　要：本研究以提取纯化的甜杏仁主要过敏原amandin为目标物,制备高亲和力的抗amandin单克隆抗体。以鼠单克隆抗体为捕获抗体、兔多克隆抗体为检测抗体,通过优化封闭液以及缓冲液pH等条件,建立双抗体夹心ELISA方法检测amandin。该方法的检出限为(0.88±0.08)ng/mL,线性范围为5.86~187.5 ng/mL。该方法与芝麻全蛋白、花生全蛋白、核桃全蛋白以及β-乳球蛋白均无交叉反应,表明特异性良好。添加回收实验中面包、饼干、巧克力回收率的范围为82.16%~112.04%,实际样品的检出限分别为(9.07±0.23)ng/mL、(7.78±0.18)ng/mL和(327.37±0.25)ng/mL,说明能够用于实际样品中amandin的准确定量分析。In this study,a high-affinity anti-amandin monoclonal antibody was prepared with the use of extracted and pur-ified amandin,the main allergen of sweet almonds,as the target.Using the mouse monoclonal antibody as the capture anti-body and the rabbit polyclonal antibody as the detection antibody,a double-antibody sandwich ELISA method was estab-lished for the detection of amandin protein by optimizing the conditions of the sealing solution and the pH of the buffer.The detection limit of this method was(0.88±0.08)ng/mL,and the linear range was 5.86~187.5 ng/mL.The method showed no cross-reactivity with whole protein of sesame,peanut,walnut,and β-lactoglobulin,indicating good specificity.The re-covery rate of bread,cookie,and chocolate in the spiked recovery test ranged from 82.16%to 112.04%,and the limits of detection(LODs)in the actual samples were(9.07±0.23)ng/mL,(7.78±0.18)ng/mL,and(327.37±0.25)ng/mL,respectively,which indicated that they can be used for the accurate quantitative analysis of amandin proteins in the actual samples.

关 键 词：杏仁过敏原 amandin 双抗夹心ELISA 

分 类 号：TS201.6[轻工技术与工程—食品科学]