| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1166876 | Analytica Chimica Acta | 2012 | 7 Pages |
For minimizing systemic experimental variation in the analysis of antibody array data, we developed a novel median-centered/IgM-tagged-internal standard (TIS) assay normalization using median-centering and TIS assay-based determination of serum IgM concentrations. We evaluated five normalization methods by analyzing correlation coefficients and coefficients of variation for six serum proteins using human serum samples from normal controls (n = 25) and patients with liver cirrhosis (n = 25) or hepatocellular carcinoma (HCC; n = 29). Median-centered normalization improved correlation coefficients, while IgM-based normalizations improved coefficients of variation. The TIS assay was more efficient, economical, and reproducible for determining IgM concentrations than enzyme-linked immunosorbent assay. Additionally, we normalized antibody array data for six serum proteins using the median-centered/IgM-TIS assay, and evaluated serum biomarkers through distribution analysis of normalized fluorescence intensities and receiver operating characteristic analyses for the diagnosis of liver cirrhosis and HCC. Apolipoprotein A-1 and a combination of alpha-fetoprotein and C-reactive protein were determined to be potential serological biomarkers for liver cirrhosis and HCC, respectively. Thus, median-centered/IgM-TIS assay normalization is a useful approach for analyzing antibody array data and evaluating serological biomarkers for the diagnosis of liver disease or cancers.
Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► We analyzed six serum proteins from 79 human serum samples using antibody arrays. ► We evaluated five normalization methods for minimizing systemic experimental variation. ► Median-centered/IgM-ELISA normalization provided was found to be optimal. ► Using this normalization, we identified potential biomarkers for liver disease. ► Potential biomarkers were verified by Western blot.
