Optimization of Rolling-Circle Amplified Protein Microarrays for Multiplexed Protein Profiling.
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.
Shao
Weiping
W
Zhou
Zhimin
Z
Laroche
Isabelle
I
Lu
Hong
H
Zong
Qiuling
Q
Patel
Dhavalkumar D.
DD
Kingsmore
Stephen
S
Piccoli
Steven P.
SP