Two-color, rolling-circle amplification on antibody microarrays for sensitive, multiplexed serum-protein measurements.

ABSTRACT: The ability to conveniently and rapidly profile a diverse set of proteins has valuable applications. In a step toward further enabling such a capability, we developed the use of rolling-circle amplification (RCA) to measure the relative levels of proteins from two serum samples, labeled with biotin and digoxigenin, respectively, that have been captured on antibody microarrays. Two-color RCA produced fluorescence up to 30-fold higher than direct-labeling and indirect-detection methods using antibody microarrays prepared on both polyacrylamide-based hydrogels and nitrocellulose. Replicate RCA measurements of multiple proteins from sets of 24 serum samples were highly reproducible and accurate. In addition, RCA enabled reproducible measurements of distinct expression profiles from lower-abund

SUBMITTER: Zhou H 

PROVIDER: S-EPMC395787 | biostudies-literature | 2004

REPOSITORIES: biostudies-literature