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

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TítuloTwo-color, rolling-circle amplification on antibody microarrays for sensitive, multiplexed serum-protein measurements.
Publication TypeJournal Article
Year of Publication2004
AutoresZhou, H, Bouwman, K, Schotanus, M, Verweij, C, Marrero, JA, Dillon, D, Costa, J, Lizardi, P, Haab, BB
JournalGenome Biol
Volume5
Issue4
PaginationR28
Date Published2004
ISSN1465-6914
Palabras claveAntibodies, Biotin, Blood Proteins, Color, Digoxigenin, Enzyme-Linked Immunosorbent Assay, Humans, Immunoassay, Liver Cirrhosis, Liver Neoplasms, Protein Array Analysis, Reproducibility of Results, Sensitivity and Specificity
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-abundance proteins that were not measurable using the other detection methods. Two-color RCA on antibody microarrays should allow the convenient acquisition of expression profiles from a great diversity of proteins for a variety of applications.

DOI10.1186/gb-2004-5-4-r28
Alternate JournalGenome Biol.
PubMed ID15059261
PubMed Central IDPMC395787
Grant ListCA81671 / CA / NCI NIH HHS / United States
P30 DK 34989 / DK / NIDDK NIH HHS / United States
UO1 CA084986 / CA / NCI NIH HHS / United States