Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, 611-0011, Uji, Japan.
Cytotechnology. 2005 Jan;47(1-3):145-9. doi: 10.1007/s10616-005-3755-6.
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.
我们最近建立了一种酶联免疫吸附测定法（ELISA），可用于测定总卵黏蛋白，无论其热变性程度如何，方法是使用一种针对卵黏蛋白碳水化合物部分的单克隆抗体（mAb）7D（Biosci Biothechnol Biochem, 68, 2490-2497, 2004）。为了改进 ELISA，我们开发了两种新方法。首先，通过使用 mAb 7D 的寡克隆鸡尾酒和另外两种具有不同表位的 mAb 作为第二抗体，将其灵敏度提高了 100 倍。其次，表明 SDS 和β-巯基乙醇等变性试剂的使用可接受用于 ELISA，只要在固相上第一抗体的稳定性范围内即可。因此，讨论了所构建的寡克隆夹心 ELISA 系统的特性，与过敏原标记有关。
