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Bioluminescent Antibodies for Point‐of‐Care Diagnostics
Author(s) -
Xue Lin,
Yu Qiuliyang,
Griss Rudolf,
Schena Alberto,
Johnsson Kai
Publication year - 2017
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201702403
Subject(s) - bioluminescence , fluorophore , luciferase , analyte , fluorescence , chemistry , antibody , antigen , förster resonance energy transfer , monoclonal antibody , biosensor , spotting , biochemistry , combinatorial chemistry , biophysics , chromatography , biology , gene , computer science , immunology , transfection , physics , quantum mechanics , artificial intelligence
We introduce a general method to transform antibodies into ratiometric, bioluminescent sensor proteins for the no‐wash quantification of analytes. Our approach is based on the genetic fusion of antibody fragments to NanoLuc luciferase and SNAP‐tag, the latter being labeled with a synthetic fluorescent competitor of the antigen. Binding of the antigen, here synthetic drugs, by the sensor displaces the tethered fluorescent competitor from the antibody and disrupts bioluminescent resonance energy transfer (BRET) between the luciferase and fluorophore. The semisynthetic sensors display a tunable response range (submicromolar to submillimolar) and large dynamic range (Δ R max >500 %), and they permit the quantification of analytes through spotting of the samples onto paper followed by analysis with a digital camera.