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使用大鼠嗜碱性白血病(RBL-1)细胞对小麦过敏原进行体外电化学检测。

In vitro electrochemical detection of wheat allergen using rat basophilic leukaemia (RBL-1) cells.

作者信息

Nakamura N, Kumazawa S, Matsunaga T

机构信息

Department of Biotechnology, Tokyo University of Agriculture and Technology, Japan.

出版信息

Appl Microbiol Biotechnol. 1995 Aug-Sep;43(4):622-5. doi: 10.1007/BF00164764.

Abstract

We have investigated an electrochemical method of detecting foods that cause an allergic reaction. Rat basophilic leukaemia (RBL-1) cells were sensitized with serum from a rat that was allergic to wheat. A sample containing the protein fraction of a food was added to the cells and incubated. The cells were immobilized on a membrane filter and attached to a basal-plane pyrolytic graphite electrode. When a potential was applied in the range 0-1.0 V relative to a saturated calomel electrode, an anodic peak current appeared at around 0.33 V. This peak current, attributed to serotonin, increased with time, and the maximum current (0.5 microA) was obtained 20-25 min of incubation. The response of the RBL-1 cells was specific to the protein fraction of wheat. The peak current increased linearly with increasing protein concentration in the range of 0.01-0.5 micrograms ml-1. These results suggest that the concentration of the protein bringing about the allergic reaction can be determined by cyclic voltammetry within 25 min. This method is more sensitive than the conventional skin tests.

摘要

我们研究了一种检测引起过敏反应食物的电化学方法。用对小麦过敏的大鼠血清使大鼠嗜碱性白血病(RBL-1)细胞致敏。将含有食物蛋白质部分的样品加入细胞中并孵育。将细胞固定在膜滤器上,并附着在基面热解石墨电极上。当相对于饱和甘汞电极施加0至1.0 V范围内的电位时,在约0.33 V处出现阳极峰值电流。这个归因于血清素的峰值电流随时间增加,孵育20至25分钟时获得最大电流(0.5微安)。RBL-1细胞的反应对小麦蛋白质部分具有特异性。在0.01至0.5微克/毫升范围内,峰值电流随蛋白质浓度增加呈线性增加。这些结果表明,引起过敏反应的蛋白质浓度可在25分钟内通过循环伏安法测定。该方法比传统的皮肤试验更灵敏。

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