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

[摘要] Protein microarray-based approaches are increasingly being used inresearch and clinical applications to either profile theexpression of proteins or screen molecular interactions. Thedevelopment of high-throughput, sensitive, convenient, andcost-effective formats for detecting proteins is a necessity forthe effective advancement of understanding disease processes. Inthis paper, we describe the generation of highly multiplexed,antibody-based, specific, and sensitive protein microarrayscoupled with rolling-circle signal amplification (RCA)technology. A total of 150 cytokines were simultaneously detectedin an RCA sandwich immunoassay format. Greater than half of theseproteins have detection sensitivities in the pg/ml range. Thevalidation of antibody microarray with human serum indicated thatRCA-based protein microarrays are a powerful tool forhigh-throughput analysis of protein expression and moleculardiagnostics.

[学科分类] 基础医学