摘要
为建立基于竞争酶联免疫分析（ELISA）的杏仁蛋白饮料中杏仁蛋白质定量方法,使用2-D电泳结合MALDI-TOF/MS鉴定了杏仁的高丰度蛋白,分离纯化了杏仁40 ku prunin蛋白α链,并以此作为抗原制备了抗杏仁蛋白单克隆抗体,使用该抗体建立了检测杏仁蛋白质的竞争酶联免疫分析方法。该方法以杏仁可溶全蛋白作为标准品,IC50为19.6μg/m L,线性范围为5.0-100μg/m L（R^2=0.990）。该方法特异性良好,和其他可食种子蛋白质无交叉反应。检测植物蛋白饮料样品的检出限为0.6 mg/m L,相对标准偏差〈10%。使用巴氏杀菌、超高温瞬时灭菌和高压灭菌的蛋白质平均回收率分别为96%、92%和81%。
The aim of the research was to develop the quantitation method of apricot kernel protein in apricot kernel protein beverage based on competitive enzyme linked immunosorbent assay（ ELISA）. We applied two-dimensional electrophoresis and MALDI-TOF / MS to identify the high-abundant protein in apricot kernel. Pru-1 subunit α-chain with a mass of 40 ku was purified and used as antigen to prepare a specific monoclonal antibody. Using the monoclonal antibody and apricot kernel soluble protein as standard,an optimized ELISA method was established linear detection range of 5. 0 to 100 μg / m L（ R^2= 0. 990）.The assay did not register cross-reactivity with other edible plant seeds. The limit of detection（ LOD） was0. 6 mg / m L for apricot kernel protein beverage,with relative standard deviation less than 10%. The average recovery rates were 96%,92% and 81% under the sterilization condition of pasteurization,ultra high temperature treated and autoclaved sterilization,respectively.
出处
《河南农业科学》
CSCD
北大核心
2016年第4期145-149,共5页
Journal of Henan Agricultural Sciences
基金
国家质检总局科技计划项目(2013QK274)
广东省质量技术监督局科技计划项目(2013CZ05)
关键词
杏仁
蛋白饮料
ELISA
定量
apricot kernel
protein beverage
enzyme linked immunosorbent assay
quantitation
