[Ru(bpy)₂(dcbpy)NHS] Labeling/Aptamer‐Based Biosensor for the Detection of Lysozyme by Increasing Sensitivity with Gold Nanoparticle Amplification

Abstract

A novel [Ru(bpy)(2)(dcbpy)NHS] labeling/aptamer-based biosensor combined with gold nanoparticle amplification for the determination of lysozyme with an electrochemiluminescence (ECL) method is presented. In this work, an aptamer, an ECL probe, gold nanoparticle amplification, and competition assay are the main protocols employed in ECL detection. With all the protocols used, an original biosensor coupled with an aptamer and [Ru(bpy)(2)(dcbpy)NHS] has been prepared. Its high selectivity and sensitivity are the main advantages over other traditional [Ru(bpy)(3)](2+) biosensors. The electrochemical impedance spectroscopy (EIS) and atomic force microscopy (AFM) characterization illustrate that this biosensor is fabricated successfully. Finally, the biosensor was applied to a displacement assay in different concentrations of lysozyme solution, and an ultrasensitive ECL signal was obtained. The ECL intensity decreased proportionally to the lysozyme concentration over the range 1.0x10(-13)-1.0x10(-8) mol L(-1) with a detection limit of 1.0x10(-13) mol L(-1). This strategy for the aptasensor opens a rapid, selective, and sensitive route for the detection of lysozyme and potentially other proteins.

Electrochemical aptasensor using the tripropylamine oxidation to probe intramolecular displacement between target and complementary nucleotide for protein array

“…The precision for five replicate determinations at different concentration level ranged from 1.4 to 4.2% (RSDs, n = 5). The acceptable relative standard deviations and quantitative recoveries imply that the present aptasensor offers a promising feature for the analytical application in complex biological samples with a simple mode over the previous biosensors for lysozyme determination Bai et al, 2008;Deng et al, 2009a;Peng et al, 2009;Du et al, 2009;Cho et al, 2006;Kawde et al, 2005).…”

“…A label-free electrochemical aptasensor for lysozyme was developed with the detection limits of 18 nM (Rodriguez and Rivas, 2009). Bai et al (2008) developed a method based on Ru[(bpy) 2 (dcbpy)]NHS labelling for the ECL determination of lysozyme. In combination with Au nanoparticle amplification, the detection limits of 0.1 pM were obtained (Bai et al, 2008).…”

“…Bai et al (2008) developed a method based on Ru[(bpy) 2 (dcbpy)]NHS labelling for the ECL determination of lysozyme. In combination with Au nanoparticle amplification, the detection limits of 0.1 pM were obtained (Bai et al, 2008). However, the synthesis of both Au nanoparticle and probe molecule is needed.…”

Analytical potential of gold nanoparticles in functional aptamer-based biosensors

“…c The schematic diagram of fabrication of ECL aptasensor for sensitive detection of thrombin based on a "sandwich" structure fabrication [106]. d Schematic representation of the preparation of solid-state ECL sensing platform for detection of adenosine [109] AuNPs for the determination of lysozyme using the ECL method was also developed by our group [107]. At first, AuNPs was self-assembled on the gold electrode.…”

Ultrasensitive and Signal‐on Electrochemiluminescence Aptasensor Using the Multi‐tris(bipyridine)ruthenium(II)‐β‐cyclodextrin Complexes

“…[25,26] Among different methods of fluorescence, [27] EC, [28] ECL [29] and colorimetry, [30,31] ECL has attracted substantial attention in the development of aptasensors because it is highly sensitive and inexpensive, and uses simple instruments. Traditionally, ECL aptasensors have used diverse luminophore-labeled aptamers, [32][33][34] which are difficult to obtain. Therefore, it is necessary to develop a simple and efficient method to fabricate aptasensors.…”