Analysis of oxidative stress biomarkers using a simultaneous competitive/non-competitive micromosaic immunoassay.
Analysis of oxidative stress biomarkers using a simultaneous competitive/non-competitive micromosaic immunoassay.
复制标题
使用同时竞争性/非竞争性微马赛克免疫分析分析氧化应激生物标志物。
DOI：
10.1016/j.aca.2009.03.003
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发表时间：
2009
影响因子：
6.2
通讯作者：
Henry,CharlesS
中科院分区：
文献类型：
--
作者：
Murphy,BrianM;Dandy,DavidS;Henry,CharlesS
免疫测定代表了从临床诊断到环境监测的许多应用的核心主力方法。在酶联免疫吸附测定（ELISA）等传统形式中，分析物是单独或以小组形式测量的。随着越来越多的生物标志物被鉴定用于疾病状态，需要开发可以同时测量多个标志物的方法。免疫亲和阵列是一种可以实现多标记筛选的化学方法。大多数阵列以竞争或非竞争形式进行，其中前者主要用于小分子，后者用于大分子。迄今为止，ELISA和免疫亲和阵列方法仅依赖于这些形式之一，而不是另一种。在这里，免疫亲和阵列方法能够同时进行竞争性和非竞争性分析，使用微镶嵌免疫分析技术产生的代谢产物和蛋白质的分析。在本报告中，三个氧化应激标志物被用作模型系统。本文所述的方法证明了3-硝基酪氨酸的同时分析，通过间接竞争性免疫测定法，而酶过氧化氢酶和超氧化物歧化酶的分析通过非竞争性夹心免疫测定法。该方法需要少于1μL的样品和45分钟完成。Logistic曲线拟合和LOD（检测限）的结合结果的统计分析，并显示出良好的协议与这些抗体-抗原系统的公布数据。
Immunoassays represent a core workhorse methodology for many applications ranging from clinical diagnostics to environmental monitoring. In traditional formats such as the enzyme linked immunosorbent assay (ELISA), analytes are measured singly or in small sets. As more biomarkers are identified for disease states, there is a need to develop methods that can measure multiple markers simultaneously. Immunoaffinity arrays are one such chemistry that can achieve multi-marker screening. Most arrays are performed in either competitive or non-competitive formats, where the former are used predominantly for small molecules and the later for macromolecules. To date, ELISA and immunoaffinity array methods have relied exclusively on one of these formats and not the other. Here an immunoaffinity array method capable of performing simultaneous competitive and non-competitive analysis generated using micromosaic immunoassay techniques is introduced for the analysis of metabolites and proteins. In this report, three markers of oxidative stress were used as a model system. The method described here demonstrates the simultaneous analysis of 3-nitrotyrosine, by indirect competitive immunoassay while the enzymes catalase and superoxide dismutase are analyzed by non-competitive sandwich immunoassay. The method requires less than 1μL sample and 45min for completion. Logistic curve fits and LOD (limits of detection) statistical analysis of the binding results are presented and show good agreement with published data for these antibody–antigen systems.