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Published on: June 9, 2017
Branched Proximity Hybridization Assay for the Quantification of Nanoscale Protein-Protein Proximity
1Laboratoire d'ImmunoRhumatologie Moléculaire, Institut national de la santé et de la recherche médicale (INSERM) UMR_S 1109, Institut thématique interdisciplinaire (ITI) de Médecine de Précision de Strasbourg, Transplantex NG, Faculté de Médecine, Fédération Hospitalo-Universitaire OMICARE, Fédération de Médecine Translationnelle de Strasbourg (FMTS), Université de Strasbourg, Strasbourg, France. jianying.yang@unistra.fr.
Abstract:
To better quantify the nanoscale protein-protein proximity, we developed a new branched proximity hybridization assay (bPHA). In this assay, oligo-coupled target protein-binding reagents, such as aptamer, nanobody, and antibodies, are used to convert the proximity of target proteins to the proximity of a pair of designed oligos. The closely positioned oligo pair then serve as a template for a maximum of 400-fold linear signal amplification through branched DNA (bDNA). The amplified bPHA signal is recorded and quantified by flow cytometer, providing high throughput, multiplexing, and single-cell resolution for nanoscale protein proximity studies.
