Normalization using a tagged-internal standard assay for analysis of antibody arrays and the evaluation of serological biomarkers for liver disease

Deok Hoon Kong, Jae Wan Jung, Keun Na, Seul Ki Jeong, Young Ki Paik, Se Hui Jung, In Bum Suh, Young Myeong Kim, Kwon Soo Ha

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)92-98
Number of pages7
JournalAnalytica Chimica Acta
Publication statusPublished - 2012 Mar 9

Bibliographical note

Funding Information:
This work was supported by the Ministry of Health and Welfare through grants from the Korea Health 21 R&D Project ( A030003 ), the National R&D Program for Cancer Control ( 1020420 ), and the Korean Ministry of Education, Science and Technology (Regional Core Research Program/Medical & Bio-Materials Research Center) .

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy


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