Development of a sensitive microarray immunoassay for the quantitative analysis of neuropeptide Y.
Development of a sensitive microarray immunoassay for the quantitative analysis of neuropeptide Y.
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开发用于神经肽 Y 定量分析的灵敏微阵列免疫测定法。
DOI：
10.1021/ac3014548
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发表时间：
2012
影响因子：
7.4
通讯作者：
Sternberg,EstherM
中科院分区：
文献类型：
--
作者：
Jia,Min;Belyavskaya,Elena;Deuster,Patricia;Sternberg,EstherM
建立了一种以抗体芯片形式检测神经肽Y(NPY)的直接竞争免疫分析方法，并将其应用于人体汗液中NPY的分析。这是首次证明抗体芯片作为一种强大的多重分析工具，可以用于NPY和潜在的其他神经肽的灵敏检测。400pg/mL二生物素化NPY和0.1 mg/mL斑点捕获抗体的检测效果最好，其灵敏度为50pg/mLNPY的线性动态范围为0.1-100 ng/mL.对人工汗液基质效应的评价表明，用芯片免疫分析技术分析人体汗液样品中的NPY时，透析是必要的。在初步应用中，在用Macroducts收集器采集的汗液样本中检测到50-210 pg/ml的NPY。这项研究表明，抗体微阵列可以用于NPY分析，人类汗液可以作为生物标记物和蛋白质组研究的有价值的样本来源，特别是当非侵入性人类样本采集更可取的时候。
A direct competitive immunoassay in an antibody microarray format was developed for the sensitive detection of neuropeptide Y (NPY) and employed in the analysis of NPY in human sweat samples. This is the first demonstration that antibody microarray, as a powerful multiplex analysis tool, can be used for the sensitive determination of NPY and potentially other neuropeptides. 400 pg/mL of dibiotinylated NPY and 0.1 mg/mL spotting capture antibody were found to offer the best performance, yielding a sensitivity of 50 pg/mL and a linear dynamic range of 0.1–100 ng/mL for NPY. Evaluation of matrix effects by using artificial sweat revealed that dialysis is necessary for analyzing NPY in human sweat samples with microarray immunoassay. In a preliminary application, 50–210 pg/mL of NPY was detected in sweat samples collected with Macroduct collectors. This study indicates that antibody microarrays can be used for NPY analysis and that human sweat could be a valuable sample source for biomarker and proteomics studies, especially when noninvasive human sample collection is preferable.