公开(公告)号：US20200157184A1

公开(公告)日：2020-05-21

申请号：US16777504

申请日：2020-01-30

申请人： The Royal Institution for the Advancement of Learning/McGill University ,  Universite de Montreal

发明人： Stéphane Alain Laporte ,  Yoon Namkung ,  Michel Bouvier ,  Christian Le Gouill ,  Mireille Hogue ,  Viktoriya Lukasheva ,  Hiroyuki Kobayashi ,  Denis Deblois ,  Étienne Durette

IPC分类号： C07K14/72 ,  G01N33/50

摘要： Bioluminescence resonance energy transfer (BRET) biosensors for assessing the intracellular localization, internalization and trafficking into cellular compartments of proteins such as receptors, and other biomolecules such as second messengers, are disclosed. These biosensors, which are dependent on the concentration/density of the BRET donor and acceptor in cellular compartments rather that specific protein-protein interactions, use a Renilla GFP/Luc BRET pair, which allows the robust and reproducible monitoring of protein trafficking/localization, with a sensitivity compatible with high-throughput screening (HTS). The use of these biosensors for various applications, including assessing/monitoring protein endocytosis, recycling and intracellular trafficking, receptor maturation/rescue by pharmacological chaperones, various endocytosis/exocytosis processes, activation/inhibition, as well as biomolecule concentration/density in different cellular compartments, is also disclosed.

公开(公告)号：US10584159B2

公开(公告)日：2020-03-10

申请号：US15512267

申请日：2015-09-21

申请人： The Royal Institution for the Advancement of Learning/McGill University ,  Université de Montréal

发明人： Stéphane Alain Laporte ,  Yoon Namkung ,  Michel Bouvier ,  Christian Le Gouill ,  Mireille Hogue ,  Viktoriya Lukasheva ,  Hiroyuki Kobayashi ,  Denis Deblois ,  Étienne Durette

IPC分类号： C07K14/72 ,  G01N33/50

摘要： Bioluminescence resonance energy transfer (BRET) biosensors for assessing the intracellular localization, internalization and trafficking into cellular compartments of proteins such as receptors, and other biomolecules such as second messengers, are disclosed. These biosensors, which are dependent on the concentration/density of the BRET donor and acceptor in cellular compartments rather that specific protein-protein interactions, use a Renilla GFP/Luc BRET pair, which allows the robust and reproducible monitoring of protein trafficking/localization, with a sensitivity compatible with high-throughput screening (HTS). The use of these biosensors for various applications, including assessing/monitoring protein endocytosis, recycling and intracellular trafficking, receptor maturation/rescue by pharmacological chaperones, various endocytosis/exocytosis processes, activation/inhibition, as well as biomolecule concentration/density in different cellular compartments, is also disclosed.