Proteomic biomarker evaluation using antibody microarrays: association between analytical methods such as microarray and ELISA.

Abstract

Objective: To evaluate the associations between analytical methods, such as microarray and enzyme-linked immunosorbent assay (ELISA); expedient cutoffs; and the lowest possible number of microarrays in analysis for target biomarker estimation in case-control studies.

Methods: This study included 321 serum specimens, gathered in different case-control studies to test for atherosclerosis and atrial fibrillation. Among them, 48 serum specimens were analyzed using microarray technology. We used ELISA and commercial kits for confirmation of the results.

Results: Three proteins-cadherin-P, neuronal nitric oxide synthase, and adenovirus fiber-were shown to have distinctly different values in the case group vs the control group. As a result, we used those proteins as the target for confirmation using our alternative analytical method. Also, these protein values represented the limiting range between the highest and lowest differences in case-control groups. The results of microarray assay were confirmed using ELISA and commercial kits in the same specimens, in which microarray profiling was performed, and also in separate large case-control groups.

Conclusions: A 1.5-fold difference in the protein content, as measured using microarray technology, was shown to be sufficient for further investigation of the candidate proteins. As few as 3 microarrays were considered sufficient for perspective evaluation of the target proteins. Microarray serum profiling, therefore, provides semiquantitative determination of protein in serum.