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Abstract: A novel biosensor for the detection of thrombin was developed by using QDs electrochemiluminescence (ECL) technique. The thiol-terminated aptamer with 15 nucleotides (probe I) was first immobilized on Au electrode, and then thrombin was imported to form the aptamer–thrombin bioaffinity complexes. Another 5′-biotin modified aptamer (29 nucleotides, probe II) was next hybridized with the combined thrombin to form a sandwich type structure. Streptavidin modified QDs (avidin–QDs) were bound to probe II via the biotin–avidin-system. The QDs ECL signal was responsive to the amount of probe II, which was indirect proportional to the combined thrombin. The ECL intensity of the biosensor increased with the increase of thrombin concentration in the range of 0–20μgmL−1. In addition, the biosensor exhibited the excellent selectivity responses and good stability toward the target analyte. [Copyright &y& Elsevier]

Quantifying the glycan expression status on cell surfaces is of vital importance for insight into the glycan function in biological processes and related diseases. Here we developed a versatile aptasensor for electrochemical quantification of cell surface glycan by taking advantage of the cell-specific aptamer, and the lectin-functionalized gold nanoparticles acting as both a glycan recognition unit and a signal amplification probe. To construct the aptasensor, amine-functionalized mucin 1 protein (MUC1) aptamer was first covalently conjugated to carboxylated-magnetic beads (MBs) using the succinimide coupling (EDC-NHS) method. On the basis of the specific recognition between aptamer and MUC1 protein that overexpressed on the surface of MCF-7 cells, the aptamer conjugated MBs showed a predominant capability for cell capture with high selectivity. Moreover, a lectin-based nanoprobe was designed by noncovalent assembly of concanavalin A (ConA) on gold nanoparticles (AuNPs). This nanoprobe incorporated the abilities of both the specific carbohydrate recognition and the signal amplification based on the gold-promoted reduction of silver ions. By coupling with electrochemical stripping analysis, the proposed sandwich-type cytosensor showed an excellent analytical performance for the ultrasensitive detection of MCF-7 cells and quantification of cell surface glycan. More importantly, taking advantage of Con A-gold nanoprobe catalyzed silver enhancement, the proposed method was further used for naked-eye tracking glycolytic inhibition in living cells. This aptasensor holds great promise as a new point-of-care diagnostic tool for analyzing glycan expression on living cells and further helps cancer diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Abstract: A sensitive colorimetric detection for biomolecules based on aptamer was described. Poly(dimethylsiloxane) (PDMS)–gold nanoparticles (AuNPs) composite film was used as a platform for immobilizing anti-target aptamer. PDMS–AuNPs composite film only covered with aptamer showed high inhibiting ability towards silver reduction, after target molecules were conjugated on the modified surface, the catalytic efficiency of AuNPs for silver reduction was increased. In this system, the darkness density of silver enhancement was applied for target quantitative measurement. Lysozyme and adenosine 5′-triphosphate (ATP) were tested as the models, quantitative measurements with imaging software or semiquantitative measurements with naked eyes were carried out in the range of 1×10−2–1μg/mL and 1×10−4–1×103 μg/mL, the volume of reagent using in each assay is 15μL or less. We speculated that aptamer-target conjugates’ inhibition ability for AuNPs’ catalytic efficiency toward silver reduction might come from charge and spatial effects. This study can offer a completely novel and relatively general approach for colorimetrical aptamer sensors with good analytical properties and potential applications. The sensor could be coupled with digital transmission of images for remote monitoring system in diagnosis, food control, and environmental analysis. [Copyright &y& Elsevier]