间接竞争ELISA检测核桃制品中核桃蛋白含量

Detection and Quantification of Walnut Protein in Walnut Products by an Indirect Competitive Enzyme Linked Immunosorbent Assay

通过核桃蛋白组分分析,确定核桃丰度蛋白。以其作为抗原免疫小鼠,制备可特异性识别核桃蛋白的单克隆抗体,建立间接竞争ELISA检测方法。筛选出一株能稳定分泌抗核桃蛋白的杂交瘤细胞株HT-1,对间接竞争ELISA方法的反应条件进行优化后,建立了检测核桃蛋白的间接竞争ELISA方法,核桃蛋白的检测线性范围在4.664.2μg/mL,IC50为17.2μg/mL,最低检测限为0.67μg/mL。抗体特异性较好,与其他的植物蛋白没有交叉反应。该方法适用于检测核桃制品中的核桃蛋白含量。

Walnut abundance proteins were find by proteomic analysis. A monoclonal antibody H1 recognizing walnut protein was selected and characterized after immunization of mice with walnut protein. One clone of the hybridoma cell stably secreting anti-walnut antibody was obtained and named HT-1. After optimization of the reactive condition, the method of indirect competitive ELISA for detection of walnut protein was established. The linear detection range was 4.664.2 μg/mL, the half inhibitory concentration（IC50） of ELISA was 17.2 μg/mL. The detection limit for walnut was 0.67 μg/mL. H1 exhibited no cross-reactivity among these food ingredients and high specificity with walnut. This method was suitable for detecting walnut protein from the walnut products.