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Optimization of Rolling-Circle Amplified Protein Microarrays for Multiplexed Protein Profiling

Author(s): Shao Weiping | Zhou Zhimin | Laroche Isabelle | Lu Hong | Zong Qiuling | Patel Dhavalkumar D. | Kingsmore Stephen | Piccoli Steven P.

Journal: Journal of Biomedicine and Biotechnology
ISSN 1110-7243

Volume: 2003;
Issue: 5;
Start page: 299;
Date: 2003;
Original page

Protein microarray-based approaches are increasingly being used in research and clinical applications to either profile the expression of proteins or screen molecular interactions. The development of high-throughput, sensitive, convenient, and cost-effective formats for detecting proteins is a necessity for the effective advancement of understanding disease processes. In this paper, we describe the generation of highly multiplexed, antibody-based, specific, and sensitive protein microarrays coupled with rolling-circle signal amplification (RCA) technology. A total of 150 cytokines were simultaneously detected in an RCA sandwich immunoassay format. Greater than half of these proteins have detection sensitivities in the pg/ml range. The validation of antibody microarray with human serum indicated that RCA-based protein microarrays are a powerful tool for high-throughput analysis of protein expression and molecular diagnostics.
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