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A Nanobody-on-Quantum Dot Displacement Assay for Rapid and Sensitive Quantification of the Epidermal Growth Factor Receptor (EGFR).
Su, Ruifang; Wu, Yu-Tang; Doulkeridou, Sofia; Qiu, Xue; Sørensen, Thomas Just; Susumu, Kimihiro; Medintz, Igor L; van Bergen En Henegouwen, Paul M P; Hildebrandt, Niko.
Afiliação
  • Su R; nanoFRET.com, Laboratoire COBRA (UMR6014 & FR3038), Université de Rouen Normandie, CNRS, INSA, Normandie Université, 76000, Rouen, France.
  • Wu YT; Nano-Science Center & Department of Chemistry, University of Copenhagen, Universitetsparken 5, 2100, Copenhagen, Denmark.
  • Doulkeridou S; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198, Gif-sur-Yvette, France.
  • Qiu X; Cell Biology, Neurobiology and Biophysics, Department of Biology, Science Faculty, Utrecht University, 3508 TB, Utrecht, The Netherlands.
  • Sørensen TJ; Princess Maxima Center, Heidelberglaan 25, 3584CS, Utrecht, The Netherlands.
  • Susumu K; Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), 91198, Gif-sur-Yvette, France.
  • Medintz IL; Key Laboratory of Marine Drug, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, 266003, Qingdao, China.
  • van Bergen En Henegouwen PMP; Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, 266237, Qingdao, China.
  • Hildebrandt N; Nano-Science Center & Department of Chemistry, University of Copenhagen, Universitetsparken 5, 2100, Copenhagen, Denmark.
Angew Chem Int Ed Engl ; 61(33): e202207797, 2022 08 15.
Article em En | MEDLINE | ID: mdl-35759268
Biosensing approaches that combine small, engineered antibodies (nanobodies) with nanoparticles are often complicated. Here, we show that nanobodies with different C-terminal tags can be efficiently attached to a range of the most widely used biocompatible semiconductor quantum dots (QDs). Direct implementation into simplified assay formats was demonstrated by designing a rapid and wash-free mix-and-measure immunoassay for the epidermal growth factor receptor (EGFR). Terbium complex (Tb)-labeled hexahistidine-tagged nanobodies were specifically displaced from QD surfaces via EGFR-nanobody binding, leading to an EGFR concentration-dependent decrease of the Tb-to-QD Förster resonance energy transfer (FRET) signal. The detection limit of 80±20 pM (16±4 ng mL-1 ) was 3-fold lower than the clinical cut-off concentration for soluble EGFR and up to 10-fold lower compared to conventional sandwich FRET assays that required a pair of different nanobodies.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Pontos Quânticos / Anticorpos de Domínio Único Tipo de estudo: Diagnostic_studies Idioma: En Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Pontos Quânticos / Anticorpos de Domínio Único Tipo de estudo: Diagnostic_studies Idioma: En Ano de publicação: 2022 Tipo de documento: Article