An aptamer folding-based sensory platform decorated with nanoparticles for simple cocaine testing.

The consumption of illicit drugs such as cannabis, cocaine, and amphetamines is still a major health and social problem, creating an abuse in adults especially. Novel techniques which estimate the drug of abuse are needed for the detection of newly revealed psychoactive drugs. Herein, we have constructed a combinatorial platform by using quantum dots (QDs) and gold nanoparticles (AuNPs) as well as a functional aptamer which selectively recognizes cocaine and its metabolite benzoylecgonine (BE). We have called it an aptamer folding-based sensory device (AFSD). For the fabrication of AFSD, QDs were initially immobilized onto the poly-L-lysine coated µ-well surfaces. Then, the AuNP-aptamer conjugates were bound to the QDs. The addition of cocaine or BE caused a change in the aptamer structure which induced the close interaction of AuNPs with the QDs. Hence, quenching of the fluorescence of QDs was observed depending on the analyte amount. The linearity of cocaine and BE was 1.0-10 nM and 1.0-25 µM, respectively. Moreover, the limits of detection for cocaine and BE were calculated as 0.138 nM and 1.66 µM. The selectivity was tested by using different interfering substances (methamphetamine, bovine serum albumin, codeine, and 3-acetamidophenol). To investigate the use of AFSD in artificial urine matrix, cocaine/BE spiked samples were applied. Also, confirmatory analyses by using high performance liquid chromatography were performed. It is shown that AFSD has a good potential for testing the cocaine abuse and can be easily adapted for detection of various addictive drugs by changing the aptamer according to desired analytes. Copyright © 2016 John Wiley & Sons, Ltd.

Polypeptide Functional Surface for the Aptamer Immobilization: Electrochemical Cocaine Biosensing.

Electroanalytical technologies as a beneficial subject of modern analytical chemistry can play an important role for abused drug analysis which is crucial for both legal and social respects. This article reports a novel aptamer-based biosensing procedure for cocaine analysis by combining the advantages of aptamers as selective recognition elements with the well-known advantages of biosensor systems such as the possibility of miniaturization and automation, easy fabrication and modification, low cost, and sensitivity. In order to construct the aptasensor platform, first, polythiophene bearing polyalanine homopeptide side chains (PT-Pala) was electrochemically coated onto the surface of an electrode and then cocaine aptamer was attached to the polymer via covalent conjugation chemistry. The stepwise modification of the surface was confirmed by electrochemical characterization. The designed biosensing system was applied for the detection of cocaine and its metabolite, benzoylecgonine (BE), which exhibited a linear correlation in the range from 2.5 up to 10 nM and 0.5 up to 50 µM for cocaine and BE, respectively. In order to expand its practical application, the proposed method was successfully tested for the analysis of synthetic biological fluids.

Polythiophene-g-poly(ethylene glycol) with Lateral Amino Groups as a Novel Matrix for Biosensor Construction.

In the ever-expanding field of conducting polymer research, functionalized graft hybrid copolymers have gained considerable interest in the biomedical engineering and biosensing applications, particularly. In the present work, a new biosensor based on conducting graft copolymer for the detection of phenolic compounds was developed. Thereby, a robust and novel material, namely "polythiophene-g-poly(ethylene glycol) with lateral amino groups" (PT-NH2-g-PEG) hybrid conducting polymer was synthesized via Suzuki condensation polymerization and characterized with (1)H NMR analysis, UV-vis spectroscopy, gel permeation chromatography (GPC) and fluorescence spectroscopy. PT-NH2-g-PEG architecture was then applied as an immobilization matrix to accomplish extended biosensing function. In a typical process, Laccase was utilized as a model enzyme for the detection of phenolic compounds. Detailed surface characterization of PT-NH2-g-PEG/Lac was performed by cyclic voltammetry, electrochemical impedance spectroscopy, atomic force microscopy, fluorescence microscopy and scanning electron microscopy measurements. Optimum pH and polymer amount were found to be pH 6.5 and 0.5 mg polymer, respectively, with the linear range of 0.0025-0.05 mM and 132.45 µA/mM sensitivity. The kinetic parameters of PT-NH2-g-PEG/Lac are 0.026 mM for Km(app) and 7.38 µA for Imax, respectively. Furthermore, the PT-NH2-g-PEG/Lac biofilm was retained 82% of its activity for 12 days indicating excellent recovery as tested with artificial wastewater.