Zangar R C, Daly D S, White A M, Servoss S L, Tan R M, Collett J R
Pacific Northwest National Laboratory, Richland, Washington 99354, USA.
J Proteome Res. 2009 Aug;8(8):3937-43. doi: 10.1021/pr900247n.
Our research group has been developing enzyme-linked immunosorbent assays (ELISA) microarray technology for the rapid and quantitative evaluation of biomarker panels. Studies using antibody microarrays are susceptible to systematic bias from the various steps in the experimental process, and these biases can mask biologically significant differences. For this reason, we have developed a calibration system that can identify and reduce systematic bias due to processing factors. Specifically, we developed a sandwich ELISA for green fluorescent protein (GFP) that is included on each chip. The GFP antigen is spiked into each biological sample or standard mixture and the resulting signal is used for calibration between chips. We developed ProMAT Calibrator, an open-source bioinformatics tool, for the rapid visualization and interpretation of the calibrator data and, if desired, data normalization. We demonstrate that data normalization using this system markedly reduces bias from processing factors. Equally useful, this calibrator system can help reveal the source of the bias, thereby facilitating the elimination of the underlying problem. ProMAT Calibrator can be downloaded at http://www.pnl.gov/statistics/ProMAT .
我们的研究小组一直在开发酶联免疫吸附测定（ELISA）微阵列技术，用于生物标志物组的快速定量评估。使用抗体微阵列的研究易受实验过程中各个步骤产生的系统偏差影响，这些偏差可能掩盖生物学上的显著差异。因此，我们开发了一种校准系统，可识别并减少因处理因素导致的系统偏差。具体而言，我们针对每个芯片上包含的绿色荧光蛋白（GFP）开发了一种夹心ELISA。将GFP抗原添加到每个生物样品或标准混合物中，所得信号用于芯片间的校准。我们开发了ProMAT校准器，这是一种开源生物信息学工具，用于快速可视化和解释校准器数据，并在需要时进行数据归一化。我们证明，使用该系统进行数据归一化可显著减少处理因素带来的偏差。同样有用的是，该校准器系统有助于揭示偏差来源，从而便于消除潜在问题。可从http://www.pnl.gov/statistics/ProMAT下载ProMAT校准器。
