Reliable enzyme-linked immunosorbent assay for the determination of soybean proteins in processed foods
Reliable enzyme-linked immunosorbent assay for the determination of soybean proteins in processed foods
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DOI：
10.1021/jf8007629
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发表时间：
2008-08-27
影响因子：
6.1
通讯作者：
Morimatsu, Fumiki
中科院分区：
文献类型：
--
作者：
Morishita, Naoki;Kamjya, Kumiko;Morimatsu, Fumiki
在过敏性食物中，大豆被认为是引起过敏性患者不良反应的食物。为了验证标记的有效性，需要建立特异、灵敏的检测方法来分析大豆蛋白。p34蛋白是大豆中的一种油体相关蛋白，被鉴定为主要的致敏蛋白之一，命名为Gly m Bd 30 K。以p34蛋白为大豆蛋白标记蛋白，采用特异性提取缓冲液，建立了一种检测和定量大豆蛋白的夹心酶联免疫吸附试验（ELISA）。所建立的双抗体夹心ELISA方法对大豆蛋白具有高度的特异性。该方法的检出限（LOD）和定量限（LOQ）分别为0.47 ng/mL（相当于食品中的0.19 μ g/g）和0.94 ng/mL（相当于食品中的0.38 μ g/g）。回收率范围为87.7%至98.7%，而批内和批间变异系数分别小于4.2%和7.5%。本研究表明，建立的ELISA方法是一种特异、准确、可靠的定量分析方法。
Among allergenic foods, soybean is known as a food causing adverse reactions in allergenic patients. To clarify the validity of labeling, the specific and sensitive detection method for the analysis of the soybean protein would be necessary. The p34 protein, originally characterized to be p34 as an oil-body associated protein in soybean, has been identified as one of the major allergenic proteins and named Gly m Bd 30K. A novel sandwich enzyme-linked immunosorbent assay (ELISA) for the detection and quantification of the soybean protein in processed foods was developed using polyclonal antibodies raised against p34 as a soybean marker protein and the specific extraction buffer for extract. The developed sandwich ELISA method was highly specific for the soybean protein. The limit of detection (LOD) and the limit of quantification (LOQ) of the developed ELISA were 0.47 ng/mL (equivalent to 0.19 mu g/g in foods) and 0.94 ng/mL (equivalent to 0.38 mu g/g in foods), respectively. The recovery ranged from 87.7 to 98.7%, whereas the intra- and interassay coefficients of variation were less than 4.2 and 7.5%, respectively. This study showed that the developed ELISA method is a specific, precise, and reliable tool for the quantitative analysis of the soybean protein in processed foods.