Streamlined circular proximity ligation assay provides high stringency and compatibility with low-affinity antibodies.

ABSTRACT: Proximity ligation assay (PLA) is a powerful tool for quantitative detection of protein biomarkers in biological fluids and tissues. Here, we present the circular proximity ligation assay (c-PLA), a highly specific protein detection method that outperforms traditional PLA in stringency, ease of use, and compatibility with low-affinity reagents. In c-PLA, two proximity probes bind to an analyte, providing a scaffolding that positions two free oligonucleotides such that they can be ligated into a circular DNA molecule. This assay format stabilizes antigen proximity probe complexes and enhances stringency by reducing the probability of random background ligation events. Circle formation also increases selectivity, since the uncircularized DNA can be removed enzymatically. We compare this meth

SUBMITTER: Jalili R 

PROVIDER: S-EPMC5798375 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature