Abstract 4625: A biotin-label-based antibody array for high-content profiling of protein expression

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Abstract Abstract: Profiling protein expression on a global scale will have significant impact on biomedical research, particularly in the discovery and development of drugs and biomarkers. Through the years, several antibody array systems have been invented and developed for multiple protein detection. However, a reliable and high-content system for protein profiling from many biological samples has yet developed. To address this problem, we have developed a high density antibody array and used this technology to uncover the potential biomarkers of ovarian cancer. In this approach, biological samples are conjugated with biotin. Antibody arrays are then used to capture the biotinylated proteins. The presence of proteins captured by the antibody chip is detected using streptavidin-conjugated fluorescent dye (Cy3 equivalent) as a reporter. The signals, which are visualized by laser scanning, are normalized using positive, negative, and internal controls. Using this biotin-label-based antibody array technology, the expression levels of 507 human, 308 mouse, 90 rat target proteins can be simultaneously detected, including cytokines, chemokines, adipokines, growth factors, angiogenic factors, proteases, soluble receptors, soluble adhesion molecules, and other proteins in a variety of samples. Most proteins can be detected at pg/ml and ng/ml levels with a CV of less than 20%. Using human biotin-based antibody arrays, we screened the serum expression profiles of 507 proteins in 47 ovarian cancer patients and 39 normal subjects. A panel of protein expression showed significant difference between normal and cancer (P<0.05). By Classification Analysis and Split-Point Score Analysis of these two groups, a small group of proteins can be used to distinguish ovarian cancer patients from normal subjects and between ovarian cancer patients and normal subjects. Our results suggest the biotin label-based antibody arrays that we have developed have great potential in applications for biomarker discovery. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4625.