Resonance energy transfer nanobiosensors based on affinity binding between apo-enzyme and its substrate.
Systems for glucose monitoring based on resonance energy transfer (RET) and competitive binding using Concanavalin A (Con A) are problematic as a result of problems of toxicity, aggregation, and irreversible binding. This paper presents an improved RET assay wherein Con A was replaced by apo-glucose oxidase (apo-GOx). The basic principle for transduction is identical to that used in assays based on Con A-dextran: a reduction in RET from fluorescein isothiocyanate (FITC) to tetramethyl rhodamine isothiocyanate (TRITC) occurs when FITC-dextran (donor) is displaced from TRITC-apo-GOx (acceptor) as a result of the competition of glucose. Fluorescence measurements confirm that the apo-GOx/dextran complexes are highly sensitive to glucose, measured as an increase in the donor peak relative to acceptor due to stepwise addition of glucose. The solution-phase assay showed strong signals and excellent repeatability, with a sensitivity of 0.0163 (ratio units)/mM over the range of 0-90 mM glucose. If properly encapsulated, these sensors can potentially be used for in vivo sensing without the concern of toxicity associated with Con A.
Chinnayelka
Swetha
S
Institute for Micromanufacturing, Louisiana Tech University, Ruston 71272, USA.
McShane
Michael J
MJ
D015374:Biosensing Techniques / Q000379:methods*; D004735:Energy Transfer / Q000502:physiology*; D049329:Nanostructures / Q000032:analysis* / Q000737:chemistry*; D011485:Protein Binding / Q000502:physiology; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013379:Substrate Specificity / Q000502:physiology