Oligoclonal enzyme-linked immunosorbent assay capable of determining the major food allergen, ovomucoid, irrespective of the degree of heat denaturation
Oligoclonal enzyme-linked immunosorbent assay capable of determining the major food allergen, ovomucoid, irrespective of the degree of heat denaturation
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DOI：
10.1007/s10616-005-3755-6
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发表时间：
2005-01-01
期刊：
影响因子：
2.2
通讯作者：
Narita, H
中科院分区：
文献类型：
--
作者：
Hirose, J;Murakami-Yamaguchi, Y;Narita, H
我们最近建立了一种酶联免疫吸附测定法（ELISA），用于测定总卵类粘蛋白，而不管其热变性程度如何，通过使用特异于卵类粘蛋白的碳水化合物部分的单克隆抗体（mAb）7 D（Biosci Biotechnol Biochem，68，2490-2497，2004）。已经开发了两种新的方法来改进ELISA。首先，通过使用mAb 7 D和具有不同表位的另外两种mAb的寡克隆混合物作为第二抗体，其灵敏度提高了100倍。第二，表明在固相上第一抗体的稳定性范围内，使用变性试剂如SDS和β-巯基乙醇进行提取对于ELISA是可接受的。因此构建的寡克隆夹心ELISA系统的性质进行了讨论与过敏原标记。
We have recently established an enzyme-linked immunosorbent assay (ELISA) for total ovomucoid determination, irrespective of the degree of its heat denaturation, by using a monoclonal antibody (mAb) 7D specific to the carbohydrate moiety of ovomucoid (Biosci Biothechnol Biochem, 68, 2490-2497, 2004). Two novel methods have been developed to improve the ELISA. First, its sensitivity was enhanced 100 times by using an oligoclonal cocktail of mAb 7D and two other mAbs with different epitopes as a second antibody. Second, it was shown that usage of denaturing reagents such as SDS and beta-mercaptoethanol for extraction was acceptable for ELISA within a range of stability of a first antibody on a solid phase. Properties of the oligoclonal sandwich ELISA system thus constructed were discussed in connection with allergen labeling.