Online Detection of Peroxidase Using 3D Printing, Active Magnetic Mixing, and Spectra Analysis.

A new method for online detection of peroxidase (POD) using 3D printing, active magnetic mixing, fluidic control, and optical detection was developed and demonstrated in this study. The proposed POD detection system consisted of a 3D printing and active magnetic mixing based fluidic chip for online catalytic reaction, an optical detector with a fluidic flow cell for quantitative determination of the final catalysate, and a single-chip microcontroller based controller for automatic control of two rotating magnetic fields and four precise peristaltic pumps. Horseradish peroxidase (HRP) was used as research model and a linear relationship between the absorbance at the characteristic wavelength of 450 nm and the concentration of HRP of 1/4-1/128 µg mL-1 was obtained as A = 0.257ln⁡(C) + 1.425 (R2 = 0.976). For the HRP spiked pork tests, the recoveries of HRP ranged from 93.5% to 110.4%, indicating that this proposed system was capable of detecting HRP in real samples. It has the potential to be extended for online detection of the activity of other enzymes and integration with ELISA method for biological and chemical analysis.

A Review on Magnetophoretic Immunoseparation.

Magnetophoresis is a motion of a magnetic or magnetizable particle induced by an inhomogeneous magnetic field in a fluid. Magnetophoretic immunoseparation, using micro- or nano-sized magnetic particles often modified by monoclonal or polyclonal antibodies for specific separation of biological or chemical targets, has shown a great potential in continuous-flow separation of cells and bacteria in clinical and biomedical fields. In this paper, the basic knowledge, key design considerations and recent developments on magnetophoretic immuno-separation of biological targets were reviewed.

A sensitive impedance biosensor based on immunomagnetic separation and urease catalysis for rapid detection of Listeria monocytogenes using an immobilization-free interdigitated array microelectrode.

In this study, we described a novel impedance biosensor combining immunomagnetic separation with urease catalysis for sensitive detection of foodborne bacteria using Listeria monocytogenes as model and an immobilization-free microelectrode as detector. The monoclonal antibodies (MAbs) were immobilized on the surface of the magnetic nanoparticles (MNPs) with the diameter of 180 nm by biotin-streptavidin system for specifically and efficiently separating Listeria cells from sample background. The polyclonal antibodies (PAbs) and the urease were modified onto the surface of the gold nanoparticles (AuNPs) with the diameter of 20 nm and the modified AuNPs were used to react with Listera to form the MNP-MAb-Listeria-PAb-AuNP-urease sandwich complexes. The urease in the complexes could catalyze the hydrolysis of the urea into ammonium carbonate and this led to an increase in the ionic strength of the media, which could be detected by the microelectrode. The magnetic separation efficiencies for L. monocytogenes at the concentrations ranging from 3.0×10(1) to 3.0×10(4) CFU/mL were over 95% for the pure cultures and over 85% for the spiked lettuce samples. The lower detection limit of this biosensor for L. monocytogenes was found to be 300 CFU/mL in both the pure cultures and the spiked lettuce samples. The microelectrode was demonstrated to be reusable for over 50 times with thorough cleaning by deionized water. This biosensor showed its potential to provide a simple, low-cost and sensitive method for rapid screening of foodborne pathogens and could be extended for detection of other biological or chemical targets.