Battery-triggered ultrasensitive electrochemiluminescence detection on microfluidic paper-based immunodevice based on dual-signal amplification strategy

Dual-signal amplification strategy for ultrasensitive electrochemiluminescence (ECL) multiplexed immunoassay on microfluidic paper-based analytical devices (μ-PADs) was demonstrated. This dual-signal amplification technique was achieved by employing graphene oxide-chitosan/gold nanoparticles (GCA) immunosensing platform and [4,4′-(2,5-dimethoxy-1,4-phenylene)bis(ethyne-2,1-diyl) dibenzoic acid] (P-acid) functionalized nanoporous silver (P-acid/NPS) signal amplification label. For further low-cost and disposable applications, battery-triggered constant-potential ECL (+1.0 V for P-acid label (vs. Ag/AgCl auxiliary electrode)) was applied on this paper-based immunodevice with the aid of a home-made voltage-tunable power device, allowing the traditional electrochemical workstation to be abandoned. We found that two tumor markers could be sequentially detected in the linear ranges of 0.003–20 and 0.001–10 ng mL⁻¹ with the detection limits down to 1.0 and 0.8 pg mL⁻¹, respectively, by simply reversing the connection mode on two working electrodes. The results exhibited excellent precision and high sensitivity of such immunoassay, and it also demonstrated that this battery-triggered ECL paper-based immunodevice could provide a rapid, simple and simultaneous multiplex immunoassay with high throughput, low-cost and low detection limits for point-of-care testing.     

Fluorescence-based immunoassay for human chorionic gonadotropin based on polyfluorene-coated silica nanoparticles and polyaniline-coated Fe3O4 nanoparticles

We report on an ultrasensitive fluorescence immunoassay for human chorionic gonadotrophin antigen (hCG). It is based on the use of silica nanoparticles coated with a copolymer (prepared from a fluorene, a phenylenediamine, and divinylbenzene; PF@SiO₂) that acts as a fluorescent label for the secondary monoclonal antibody to β-hCG antigen. In parallel, Fe₃O₄ nanoparticles were coated with polyaniline, and these magnetic particles (Fe₃O₄@PANI) served as a solid support for the primary monoclonal antibody to β-hCG antigen. The PF@SiO₂ exhibited strong fluorescence and good dispersibility in water. A fluorescence sandwich immunoassay was developed that enables hCG concentrations to be determined in the 0.01–100 ng·mL^?1 concentration range, with a detection limit of 3 pg·mL^?1.

Molecular simulation of the interaction between novel type rhodanine derivative probe and bovine serum albumin

The interaction between 3-(4′-methylphenyl)-5-(4′-methyl-2′-sulfophenylazo) rhodanine (M4MRASP) and bovine serum albumin (BSA) was studied by using spectrofluorimetry. It was shown in fluorescence spectrums that the quenching mechanism of BSA by M4MRASP was a static quenching. Meanwhile, the binding constant and binding site numbers were calculated. The action distance (r = 8.03 nm) and energy transfer efficiency (E = 0.12) between donor (BSA) and acceptor (M4MRASP) were obtained according to the theory of Förster non-radiation energy transfer. The effect of M4MRASP on the conformation of BSA was further analyzed by using synchronous fluorescence spectrometry. A new model of the interaction between small organic molecule and biomacromolecule was established. The results offered a reference for the studies on the biological effects and action mechanism of small molecule with protein.     

Highly sensitive hybridization assay using the electrochemiluminescence of an ITO electrode, CdTe quantum dots functionalized with hierarchical nanoporous PtFe nanoparticles, and magnetic graphene nanosheets

We report on a disposable microdevice suitable for sandwich-type electrochemiluminescence (ECL) detection of DNA. The method is making use of CdTe quantum dots functionalized with hierarchical nanoporous PtFe (CdTe@PtFe) nanoparticles and with magnetic graphene nanosheets. The latter were selected as carriers for the capture DNA due to their excellent biomagnetic separation capability and electrical properties. The CdTe@PtFe nanoparticles were used to label the signal DNA which resulted in distinctly enhanced ECL owing to the large specific surface area and good electrical conductivity of the PtFe alloy. A DNA sensor was constructed on a disk-shaped indium tin oxide electrode that was fabricated via etching. Under optimal conditions, the biosensor responds linearly to DNA in the 0.02 fM to 5000 fM concentration range, with a detection limit as low as 15 aM. The electrode is regenerable. The method displays excellent specificity, extremely good sensitivity, and is highly reproducible.

Highly selective molecular recognition and high throughput detection of melamine based on molecularly imprinted sol-gel film

Multimode reader has been generally applied in immunoassay, and in the proposed paper, the 96 well micro-plate was modified with molecularly imprinted melamine sol-gel film, based on which the highly selective and high throughput detection of melamine was achieved. Melamine was imprinted into silica sol-gel films directly using phenyltrimethoxysilane and methyltrimethoxysilane as functionalized organosilicon precursors. The binding characteristic of the imprinted film to melamine was evaluated by equilibrium binding experiments and the morphology was studied by scanning electronic microscope (SEM). Scatchard analysis was carried out to estimate the binding parameters of the imprinted film. The proposed method exhibited excellent selectivity because of specific recognition of MM by molecularly imprinted film. Under the optimum conditions, the chemiluminescence (CL) intensity had a linear relationship against the concentration of melamine over the range of 0.1-50 μg mL⁻¹ with a lower detection limit of 0.02 μg mL⁻¹.     

Paper-based electrochemiluminescent 3D immunodevice for lab-on-paper, specific, and sensitive point-of-care testing

Recent research on microfluidic paper-based analytical devices (μPADs) has shown that paper has great potential for the fabrication of low-cost diagnostic devices for healthcare and environmental monitoring applications. Herein, electrochemiluminescence (ECL) was introduced for the first time into μPADs that were based on screen-printed paper-electrodes. To further perform high-specificity, high-performance, and high-sensitivity ECL on μPADs for point-of-care testing (POCT), ECL immunoassay capabilities were introduced into a wax-patterned 3D paper-based ECL device, which was characterized by SEM, contact-angle measurement, and electrochemical impedance spectroscopy. With the aid of a home-made device-holder, the ECL reaction was triggered at room temperature. By using a typical tris(bipyridine)ruthenium-tri-n-propylamine ECL system, this paper-based ECL 3D immunodevice was applied to the diagnosis of carcinoembryonic antigens in real clinical serum samples. This contribution further expands the number of sensitive and specific detection modes of μPADs.     

Determination of sibutramine with a new sensor based on luminol electrochemiluminescence

A flow injection electrochemiluminescence (FI-ECL) analysis method for the determination of sibutramine in the presence of luminol was studied under conventional cyclic voltammetry in alkaline Na₂CO₃-NaHCO₃ buffer solution (pH 8.0-12.0). This method is based on the enhanced ECL of luminol-sibutramine. Meanwhile, in order to overcome the drawbacks of conventional cells, a FI cell was designed, which is reusable and has a great improvement in sensitivity and selectivity for ECL analysis. Under the optimal experimental conditions, the enhanced ECL intensity was linearly related to the concentration of sibutramine in the range 1.0×10⁻⁸-1.0×10⁻⁶ g mL⁻¹ with a detection limit of 2.48×10⁻⁹ g mL⁻¹ and a correlation coefficient (R) of 0.9995. The relative standard deviation (RSD) for 1.0×10⁻⁷ g mL⁻¹ samples was 2.1% (n=11). The possible mechanism discussed. The proposed FI-ECL method has been successfully applied to the determination of sibutramine in diet pill samples.