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

Proximity based detection methods are invaluable tools in the field of molecular biology, increasing selectivity and allowing for analysis of protein interactions. ProxHCR utilizes pairs of antibodies labelled with oligonucleotides to probe for proximal binding and to initiate a hybridization chain...

Full description

Saved in:
Bibliographic Details
Published inMolecular systems design & engineering Vol. 4; no. 5; pp. 158 - 165
Main Authors Leino, Mattias, Heldin, Johan, Sander, Marie Rubin, Kermpatsou, Despoina, Raykova, Doroteya, Koos, Björn, Söderberg, Ola
Format Journal Article
LanguageEnglish
Published Cambridge Royal Society of Chemistry 2019
Subjects
Antibodies
Hybridization
Molecular biology
Oligonucleotides
Optimization
Selectivity
Temperature dependence
Online AccessGet full text

Cover

More Information
Summary:Proximity based detection methods are invaluable tools in the field of molecular biology, increasing selectivity and allowing for analysis of protein interactions. ProxHCR utilizes pairs of antibodies labelled with oligonucleotides 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 dependent on salt concentrations and temperature. Herein we have redesigned the proxHCR system to increase the performance and to reduce dependency on temperature and salt concentrations. The new oligonucleotides provide an increased signal when performed at physiological salt concentrations and in room temperature. Optimization of proximity-dependent initiation of hybridization chain reaction for use at varying concentrations of salt and temperature.
ISSN:2058-9689
2058-9689
DOI:10.1039/c9me00079h