Donghoon Kwon

3D-Printed Microfluidic Device for the Detection of Pathogenic Bacteria Using Size-based Separation in Helical Channel with Trapezoid Cross-Section

A facile method has been developed to detect pathogenic bacteria using magnetic nanoparticle clusters (MNCs) and a 3D-printed helical microchannel. Antibody-functionalized MNCs were used to capture E. coli (EC) bacteria in milk, and the free MNCs and MNC-EC complexes were separated from the milk using a permanent magnet. The free MNCs and MNC-EC complexes were dispersed in a buffer solution, then the solution was injected into a helical microchannel device with or without a sheath flow. The MNC-EC complexes were separated from the free MNCs via the Dean drag force and lift force, and the separation was facilitated in the presence of a sheath flow. The concentration of the E. coli bacteria was determined using a light absorption spectrometer, and the limit of detection was found to be 10 cfu/mL in buffer solution and 100 cfu/mL in milk.

Ultrarapid detection of pathogenic bacteria using a 3D immunomagnetic flow assay.

We developed a novel 3D immunomagnetic flow assay for the rapid detection of pathogenic bacteria in a large-volume food sample. Antibody-functionalized magnetic nanoparticle clusters (AbMNCs) were magnetically immobilized on the surfaces of a 3D-printed cylindrical microchannel. The injection of a Salmonella-spiked sample solution into the microchannel produced instant binding between the AbMNCs and the Salmonella bacteria due to their efficient collisions. Nearly perfect capture of the AbMNCs and AbMNCs-Salmonella complexes was achieved under a high flow rate by stacking permanent magnets with spacers inside the cylindrical separator to maximize the magnetic force. The concentration of the bacteria in solution was determined using ATP luminescence measurements. The detection limit was better than 10 cfu/mL, and the overall assay time, including the binding, rinsing, and detection steps for a 10 mL sample took less than 3 min. To our knowledge, the 3D immunomagnetic flow assay described here provides the fastest high-sensitivity, high-capacity method for the detection of pathogenic bacteria.

Highly sensitive diagnostic assay for the detection of protein biomarkers using microresonators and multifunctional nanoparticles.

We developed a novel gravimetric immunoassay for sensitive detection of multiple protein biomarkers using silicon microcantilever arrays and multifunctional hybrid nanoparticles. Magnetic-photocatalytic hybrid nanoparticles with a highly crystalline TiO(2) shell were synthesized using a solvothermal reaction without a calcination process. After functionalizing the hybrid nanoparticles and silicon cantilevers with antibodies, the nanoparticles were used to magnetically separate target biomarkers from human serum. Frequency changes of the microcantilevers due to the binding of the nanoparticles were measured using a dip-and-dry method. Frequency changes were further amplified using a photocatalytic silver reduction reaction. Several biomarkers, including interleukin-6, interferon-γ, and alpha-fetoprotein, were selectively detected using arrays of eight silicon microcantilevers. The detection limit of this assay was ∼0.1 pg/mL, which is superior to the clinical threshold of the biomarkers.

Silicon nanowire biosensors for detection of cardiac troponin I (cTnI) with high sensitivity

We have demonstrated highly sensitive and label-free detection of cardiac troponin I (cTnI), a biomarker for diagnosis of acute myocardial infarction, using silicon nanowire field-effect transistors. A honeycomb-like structure is utilized for nanowire configuration to offer improved electrical performance and increased sensing area. The fabricated devices show n-type behavior with a relatively high ON-OFF current ratio, small sub-threshold swing and low gate leakage current. Monoclonal antibodies for cTnI were covalently immobilized on the nanowire surface and the attachment of antibodies is clearly visualized by atomic force microscope. The sensitivity with various concentrations of buffer solution was also investigated in order to determine the optimal buffer condition. The devices exhibit highest sensitivity under buffer solutions with low ion concentration. In addition, the detection limit of the sensor is as low as ~5 pg/mL, the lowest reported in the literature to date and nearly an order of magnitude smaller than the suggested threshold limit. The fabricated devices demonstrate a good selectivity for detecting cTnI.

A facile and sensitive immunoassay for the detection of alpha-fetoprotein using gold-coated magnetic nanoparticle clusters and dynamic light scattering

A facile and sensitive immunoassay protocol for the detection of alpha-fetoprotein (AFP) was developed using gold-coated iron oxide magnetic nanoclusters and dynamic light scattering (DLS) methods. The increase in the average particle size due to AFP-mediated aggregation was measured using DLS, and the detection limit was better than 0.01 ng mL(-1).

Colorimetric detection of pathogenic bacteria using platinum-coated magnetic nanoparticle clusters and magnetophoretic chromatography

A colorimetric method that uses platinum-coated magnetic nanoparticle clusters (Pt/MNCs) and magnetophoretic chromatography is developed to detect pathogenic bacteria. Half-fragments of monoclonal Escherichia coli O157:H7 (EC) antibodies were functionalized to Pt/MNCs and used to capture E. coli bacteria in milk. After magnetic separation of free Pt/MNCs and Pt/MNC-EC complexes from the milk, a precision pipette was used to imbibe the E. coli-containing solution, then a viscous polyethylene glycol solution. Due to difference in viscosities, the solutions separate into two liquid layers inside the pipette tip. The Pt/MNC-EC complexes were separated from the free Pt/MNCs by applying an external magnetic field, then added to a tetramethylbenzidine (TMB) solution. Catalytic oxidation of TMB by Pt produced color changes of the solution, which enabled identification of the presence of 10 cfu mL(-1) E. coli bacteria with the naked eye. The total assay time including separation, binding and detection was 30 min.

Facile and Sensitive Method for Detecting Cardiac Markers using Ubiquitous pH Meters

A sensitive and easy method was developed for the detection of the cardiac marker troponin I using magnetic immunoassay and ubiquitous pH meters. Monoclonal antibody-functionalized Fe3O4 magnetic nanoparticle clusters (MNCs) were synthesized to capture troponin in human serum, and MNC-troponin complexes were magnetically isolated using a permanent magnet. These complexes were subsequently conjugated to polyclonal antibody-functionalized acetylcholinesterase (AchE) and dispersed in acetylcholine (Ach) solution. As the Ach was hydrolyzed to choline and acetic acid, the pH of the solution decreased, and the resulting pH change was measured in real time using a pH meter. The sensitivity of detection of this assay was found to be 10 pg/mL of troponin in human serum after 10 min of the hydrolysis reaction. Further, the pH change could be determined with the naked eye from the color change of a pH indicator strip.

Magnetophoretic Chromatography for the Detection of Pathogenic Bacteria with the Naked Eye

A facile and sensitive analytical method that uses gold-coated magnetic nanoparticle clusters (Au/MNCs) and magnetophoretic chromatography with a precision pipet has been developed for the detection of Salmonella bacteria. Antibody-conjugated Au/MNCs are used to capture the Salmonella bacteria in milk and are then separated from the milk by applying an external magnetic field. The Salmonella-containing solution is sucked into a precision pipet tip to which a viscous polymer solution is then added. Once the magnetophoretic chromatography process has been carried out for 10 min, the presence of 100 cfu/mL Salmonella bacteria can be detected with the naked eye because the bacteria have become concentrated at the narrow pipet tip. The performance of this method was evaluated by using dynamic light scattering and light absorption spectroscopy.

Facile detection of Troponin I using dendritic platinum nanoparticles and capillary tube indicators

A facile method was developed for the detection of Troponin I (Tnl) using dendritic platinum nanoparticles and capillary tube indicators. Dendritic platinum nanoparticles were functionalized with TnI antibodies, which were used to capture TnI in human serum. The captured TnI was conjugated to the inner surface of a glass vial, to which a hydrogen peroxide (H2O2) solution was added. After sealing the glass vial with a screw cap containing a silicon septum, a capillary tube containing a drop of ink was inserted through the septum. The catalytic dissociation of H2O2 to water and oxygen increased the pressure inside the glass vial and raised the ink level in the capillary tube. The ink level increased with the platinum nanoparticle concentration, which is proportional to the TnI concentration. The sensitivity of this assay for TnI in human serum after a 5-min dissociation reaction, detected with the naked eye, was 0.1 ng/mL, which was better than the sensitivity of the conventional colorimetric method using the TMB oxidation reaction under the same experimental conditions. A control experiment using alpha-fetoprotein, interleukin-5, and C-reactive protein revealed that the developed method was highly selective for the detection of.

Detection of Salmonella bacteria in milk using gold-coated magnetic nanoparticle clusters and lateral flow filters

A novel method was developed for the detection of Salmonella bacteria using gold-coated magnetic nanoparticle clusters (Au/MNCs) and lateral flow filters. Unlike a conventional lateral flow immunoassay (LFA) membrane, where antibodies are immobilized on test and control lines to observe a signal, the test line in the new method is formed simply by pressing a nitrocellulose membrane to decrease its thickness. Half-antibody fragment-functionalized Au/MNCs were used to capture Salmonella in milk and then dispersed in a small volume of buffer solution in which one end of the lateral flow filter was immersed. The free Au/MNCs rose to the pressed test line, while the Salmonella–Au/MNC complexes remained in the solution because they were too large to pass through the pores inside the lateral flow filter. The flow of free Au/MNCs was blocked at the test line because of the reduced pore size, and their accumulation caused the test line to darken. The color of the test line was inversely proportional to the Salmonella concentration, and the limit of detection for Salmonella in milk was determined to be 103 CFU mL−1 after image processing. A control test using Escherichia coli, Staphylococcus aureus, and Vibrio parahaemolyticus revealed that the new lateral flow filtration method with Au/MNCs is highly selective for the detection of Salmonella.