Interfacial Chemistry and the Design of Solid-Phase Nucleic Acid Hybridization Assays Using Immobilized Quantum Dots as Donors in Fluorescence Resonance Energy Transfer

被引:29
作者
Algar, W. Russ [1 ]
Krull, Ulrich J. [1 ]
机构
[1] Univ Toronto, Chem Sensors Grp, Dept Chem & Phys Sci, Mississauga, ON L5L 1C6, Canada
来源
SENSORS | 2011年 / 11卷 / 06期
基金
加拿大自然科学与工程研究理事会;
关键词
active area; biosensor; DNA; fluorescence resonance energy transfer; immobilization; regeneration; signal enhancement; quantum dots; DNA; OLIGONUCLEOTIDES; TRANSDUCTION; BIOANALYSIS; SELECTIVITY; ADSORPTION; STABILITY; FORMAMIDE; PROTEINS; DENSITY;
D O I
10.3390/s110606214
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The use of quantum dots (QDs) as donors in fluorescence resonance energy transfer (FRET) offer several advantages for the development of multiplexed solid-phase QD-FRET nucleic acid hybridization assays. Designs for multiplexing have been demonstrated, but important challenges remain in the optimization of these systems. In this work, we identify several strategies based on the design of interfacial chemistry for improving sensitivity, obtaining lower limits of detection (LOD) and enabling the regeneration and reuse of solid-phase QD-FRET hybridization assays. FRET-sensitized emission from acceptor dyes associated with hybridization events at immobilized QD donors provides the analytical signal in these assays. The minimization of active sensing area reduces background from QD donor PL and allows the resolution of smaller amounts of acceptor emission, thus lowering the LOD. The association of multiple acceptor dyes with each hybridization event can enhance FRET efficiency, thereby improving sensitivity. Many previous studies have used interfacial protein layers to generate selectivity; however, transient destabilization of these layers is shown to prevent efficient regeneration. To this end, we report a protein-free interfacial chemistry and demonstrate the specific detection of as little as 2 pmol of target, as well as an improved capacity for regeneration.
引用
收藏
页码:6214 / 6236
页数:23
相关论文
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