摘要
Abstract
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ELISAKey words
almond allergen/amandin/double antibody sandwich ELISA分类
轻工业
