Transcription Factor Based Small-Molecule Sensing with a Rapid Cell Phone Enabled Fluorescent Bead Assay

被引：22

作者：

Chern, Margaret ^{[1
]}

Garden, Padric M. ^{[2
]}

Baer, R. C. ^{[3
]}

Galagan, James E. ^{[2
,3
,4
]}

Dennis, Allison M. ^{[1
,2
]}

机构：

[1] Boston Univ, Div Mat Sci & Engn, Boston, MA 02215 USA

[2] Boston Univ, Dept Biomed Engn, Boston, MA 02215 USA

[3] Boston Univ, Dept Microbiol, Boston, MA 02215 USA

[4] Boston Univ, Natl Emerging Infect Dis Labs, Boston, MA 02215 USA

来源：

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | 2020年 / 59卷 / 48期

基金：

美国国家科学基金会; 美国国家卫生研究院;

关键词：

biosensors; DNA; fluorescence; quantum dots; transcription factors; QUANTUM DOTS; BIOSENSORS; SENSITIVITY; IMMUNOASSAY; RECOGNITION; SMARTPHONE; BINDING; TETR;

D O I：

10.1002/anie.202007575

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Recently, allosteric transcription factors (TFs) were identified as a novel class of biorecognition elements for in vitro sensing, whereby an indicator of the differential binding affinity between a TF and its cognate DNA exhibits dose-dependent responsivity to an analyte. Described is a modular bead-based biosensor design that can be applied to such TF-DNA-analyte systems. DNA-functionalized beads enable efficient mixing and spatial separation, while TF-labeled semiconductor quantum dots serve as bright fluorescent indicators of the TF-DNA bound (on bead) and unbound states. The prototype sensor for derivatives of the antibiotic tetracycline exhibits nanomolar sensitivity with visual detection of bead fluorescence. Facile changes to the sensor enable sensor response tuning without necessitating changes to the biomolecular affinities. Assay components self-assemble, and readout by eye or digital camera is possible within 5 minutes of analyte addition, making sensor use facile, rapid, and instrument-free.

引用

页码：21597 / 21602

页数：6

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