Journal
ANALYTICAL BIOCHEMISTRY
Volume 380, Issue 2, Pages 164-173Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2008.05.018
Keywords
aptamer; real-time PCR; aptamer beacon; conformation-switching aptamer; SELEX; diagnostics
Funding
- NIBIB NIH HHS [R01 EB003424-03, R01 EB003424] Funding Source: Medline
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We have developed a novel method that uses conformation-switching aptamers for real-time PCR analysis of protein analytes. The aptamers have been designed so that they assume one secondary structure in the absence of a protein analyte and a different secondary structure in the presence of a protein such as thrombin or platelet-derived growth factor (PDGF). The protein-bound structure in turn assembles a ligation junction for the addition of a real-time PCR primer. Protein concentrations could be specifically detected into the picomolar range, even in the presence of cell lysates. The method has advantages relative to both immunoPCR (because no signal is produced by background binding) and the proximity ligation assay (PLA) (because only one epitope, rather than two epitopes, on a protein surface must be bound). (c) 2008 Elsevier Inc. All rights reserved.
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