期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 56, 期 25, 页码 7112-7116出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201702403
关键词
antibodies; bioluminescence; point-of-care monitoring; sensors; therapeutic drug monitoring
资金
- Swiss National Science Foundation
- NCCR Chemical Biology
- EPFL
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 (Delta R-max>500%), and they permit the quantification of analytes through spotting of the samples onto paper followed by analysis with a digital camera.
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