Optimization of proximity-dependent initiation of hybridization chain reaction for improved performance

摘要

Proximity based detection methods are invaluable tools in the field of molecular biology, increasing selec- tivity and allowing for analysis of protein interactions. ProxHCR utilizes pairs of antibodies labelled with oli- gonucleotides to probe for proximal binding and to initiate a hybridization chain reaction (HCR) to generate an amplified detection signal. As HCR is based upon hybridization of DNA hairpins, the performance is de- pendent on salt concentrations and temperature. Herein we have redesigned the proxHCR system to in- crease the performance and to reduce dependency on temperature and salt concentrations. The new oli- gonucleotides provide an increased signal when performed at physiological salt concentrations and in room temperature.