Proximity-dependent initiation of hybridization chain reaction

Björn Koos, Gaëlle Cane, Karin Grannas, Liza Löf, Linda Arngården, Johan Heldin, Carl-Magnus Clausson, Axel Klaesson, M. Karoliina Hirvonen, Felipe M. S. de Oliveira, Vladimir O. Talibov, Nhan T. Pham, Manfred Auer, U. Helena Danielson, Johannes Haybaeck, Masood Kamali-Moghaddam and Ola Söderberg ()
Additional contact information
Björn Koos: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
Gaëlle Cane: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
Karin Grannas: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
Liza Löf: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
Linda Arngården: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
Johan Heldin: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
Carl-Magnus Clausson: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
Axel Klaesson: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
M. Karoliina Hirvonen: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
Felipe M. S. de Oliveira: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
Vladimir O. Talibov: Box 256, Uppsala University
Nhan T. Pham: School of Biological Sciences and School of Biomedical Sciences, University of Edinburgh, C H Waddington Building, Max Born Cresent, Kings Buildings, Edinburgh EH9 3BF, UK
Manfred Auer: School of Biological Sciences and School of Biomedical Sciences, University of Edinburgh, C H Waddington Building, Max Born Cresent, Kings Buildings, Edinburgh EH9 3BF, UK
U. Helena Danielson: Box 256, Uppsala University
Johannes Haybaeck: Institute of Pathology, Medical University of Graz
Masood Kamali-Moghaddam: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center
Ola Söderberg: Uppsala University, Genetics and Pathology, Science for Life Laboratory, Biomedical center

Nature Communications, 2015, vol. 6, issue 1, 1-10

Abstract: Abstract Sensitive detection of protein interactions and post-translational modifications of native proteins is a challenge for research and diagnostic purposes. A method for this, which could be used in point-of-care devices and high-throughput screening, should be reliable, cost effective and robust. To achieve this, here we design a method (proxHCR) that combines the need for proximal binding with hybridization chain reaction (HCR) for signal amplification. When two oligonucleotide hairpins conjugated to antibodies bind in close proximity, they can be activated to reveal an initiator sequence. This starts a chain reaction of hybridization events between a pair of fluorophore-labelled oligonucleotide hairpins, generating a fluorescent product. In conclusion, we show the applicability of the proxHCR method for the detection of protein interactions and posttranslational modifications in microscopy and flow cytometry. As no enzymes are needed, proxHCR may be an inexpensive and robust alternative to proximity ligation assays.

Date: 2015
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DOI: 10.1038/ncomms8294

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