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Journal of Biomedicine and Biotechnology
Volume 2003 (2003), Issue 5, Pages 299-307
http://dx.doi.org/10.1155/S1110724303209268
Research article

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

1Molecular Staging, Inc, Suite 701, 300 George Street, New Haven 06511, CT, USA
2Thurston Arthritis Research Center, Department of Medicine, University of North Carolina at Chapel Hill, CB# 7280, 3330 Thurston Building, Chapel Hill 27599-7280, NC, USA

Received 13 August 2002; Accepted 10 December 2002

Copyright © 2003 Hindawi Publishing Corporation. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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.