Surface Immobilized Nucleic Acid–Transcription Factor Quantum Dots for Biosensing
Abstract
Immobilization of biosensors on surfaces is a key step toward development of devices for real‐world applications. Here the preparation, characterization, and evaluation of a surface‐bound transcription factor–nucleic acid complex for analyte detection as an alternative to conventional systems employing aptamers or antibodies are described. The sensor consists of a gold surface modified with thiolated Cy5 fluorophore‐labeled DNA and an allosteric transcription factor (TetR) linked to a quantum dot (QD). Upon addition of anhydrotetracycline (aTc)—the analyte—the TetR‐QDs release from the surface‐bound DNA, resulting in loss of the Förster resonance energy transfer signal. The sensor responds in a dose‐dependent manner over the relevant range of 0–200 µm aTc with a limit of detection of 80 nm. The fabrication of the sensor and the subsequent real‐time quantitative measurements establish a framework for the design of future surface‐bound, affinity‐based biosensors using allosteric transcription factors for molecular recognition.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jul 21, 2020
- Source ID
- 10.1002/adhm.202000403
Entities
People
- Allison M Dennis
- Catherine M. Klapperich
- Chloé Grazon
- James Galagan
- Margaret Chern
- Mark W Grinstaff
- Mingfu Chen
- Nitinun Varongchayakul
- R. C. Baer
- Sébastien Lecommandoux
- Thuy T. Nguyen
Organizations
- Biomedical Engineering Department, University of Michigan
- Boston University
- Defense Advanced Research Projects Agency
- Horizon 2020
- National Institutes of Health