• Journal of Magnetics
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• v.19 no.1
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• pp.10-14
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• 2014

In this study, we fabricate an integrated microfluidic chip with a planar Hall effect (PHE) sensor for single magnetic bead detection. The PHE sensor was constructed with a junction size of $10{\mu}m{\times}10{\mu}m$ using a trilayer structure of Ta(3 nm)/NiFe(10 nm)/Cu(1.2 nm)/IrMn(10 nm)/Ta(3 nm). The sensitivity of the PHE sensor was 19.86 ${\mu}V/Oe$. A diameter of 8.18 ${\mu}m$ magnetic beads was used, of which the saturation magnetization was ~2.1 emu/g. The magnetic susceptibility ${\chi}$ of these magnetic beads was calculated to be ~0.14. The diluted magnetic beads solution was introduced to the microfluidic channel attributing a single bead flow and simultaneously the PHE sensor voltage was measured to be 0.35 ${\mu}V$. The integrated microchip was able to detect a magnetic moment of $1.98{\times}10^{-10}$ emu.

• Journal of Magnetics
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• v.12 no.1
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• pp.40-44
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• 2007

The Planar Hall effect in a spin valve structure has been applied as a biosensor being capable of detecting $Dynabeads^{(R)}$ M-280. The sensor performance was tested under the application of a DC magnetic field where the output signals were obtained from a nanovoltmeter. The sensor with the pattern size of $50{\times}100{\mu}m^2$ has produced high sensitivity; especially, the real-time profiles by using that sensor revealed significant performance at external applied magnetic field of around 7.0 Oe with the resolution of 0.04 beads per $\mu m^2$. Finally, a successful array including 24 patterns with the single sensor size of $3{\times}3{\mu}m^2$ has shown the uniform and stable signals for single magnetic bead detection. The comparison of this sensor signal with the others has proved feasibility for biosensor application. This, connecting with the advantages of more stable and high signal to noise of PHR sensor's behaviors, can be used to detect the biomolecules and provide a vehicle for detection and study of other molecular interaction.

• Proceedings of the Korean Magnestics Society Conference
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• 2003.06a
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• pp.48-49
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• 2003

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.6
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• pp.948-955
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• 1997

Immunomagnetic separation of virus coupled with .reverse transcription-polymerase chain reaction (IMS-PCR) was performed with infectious hematopoietic necrosis virus (IHNV). A DNA fragment of expected size was synthesized in the RT-PCR with total RNA extracted from IHNV inoculated CHSE-214. In a SDS-PAGE analysis, a protein band of over 70kDa was detected from non-infected cells and cells inoculated with IHNV and infectious pancreatic necrosis virus (IPNV). This protein was detected in the Western blot analysis probably because of non-specific reaction to monoclonal antibody against IHNV nucleocapsid protein. In the immunomagnetic separation, magnetic beads coated with monoclonal antibody against the IHNV nucleocapsid protein was incubated with supernatant from IHNV inoculated CHSE-214 cells. During this process, the non-specifically reacting protein could be removed by washing the magnetic bead with PBS in the presence of an external magnetic field, and viral proteins were detected from the remaining, cleaned magnetic beads. It was necessary to extract viral RNA from the captured virus particles before RT-PCR, and no DNA product was detected when the captured virus was only heated 5 min at $95^{\circ}C$. A PCR-product of expected size was synthesized from IMS-PCR with magnetic beads double coated either by goat anti-mouse IgG antibody -monoclonal antibody or streptavidin - biotin conjugated monoclonal antibody.

• Food Science of Animal Resources
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• v.38 no.4
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• pp.727-736
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• 2018

In this study, an immuno-magnetic bead (IMB)-based assay was developed to simultaneously detect Escherichia coli, Staphylococcus aureus, and Salmonella spp. and was tested in four animal-derived foods: beef, ham, egg, and ricotta cheese. The IMB-based assay exhibited good specificity by binding to five E. coli serotypes [capture efficiency (CE) average (avg.) 90.4%], five S. aureus strains (CE avg. 91.4%), and five Salmonella serotypes (CE avg. 95.4%) but not binding to non-target bacteria (CE<10%). Furthermore, the assay detected all three pathogens with a detection limit of 10 CFU/g without the need for enrichment or additional platforms. Since the results demonstrated that the IMB-based assay can effectively separate and enrich target bacteria from a variety of animal-derived food matrixes, the assay exhibits good specificity for potential use in providing rapid, immunological, presumptive identification of pathogenic bacteria.

• Proceedings of the Korean Magnestics Society Conference
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• 2015.05a
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• pp.161-163
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• 2015

The micro device, coil, and channel for the biosensor integrated with the GMR-SV device based on the antiferromagnetic IrMn layer was fabricated by the light lithography process. When RBCs coupled with several magnetic beads with a diameter of $1{\mu}m$ passed on the micro channel, the movement of RBC + ${\mu}Beads$ is controlled by the electrical AC input signal. The RBC + ${\mu}Beads$ having a micro-magnetic field captured above the GMR-SV device is changed as the output signals for detection status. From these results, the GMR-SV device having the width magnitude of a few micron size can be applied as the biosensor for the analysis of a new magnetic property as the membrane's deformation of RBC coupled to magnetic beads.

• Food Science and Biotechnology
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• v.17 no.3
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• pp.651-654
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• 2008

In order to enhance the efficacy of norovirus detection by reverse transcriptase-polymerase chain reaction (RT-PCR) and nested PCR, this study developed a norovirus mRNA concentration method using poly oligo dT-conjugated magnetic beads. An efficient norovirus detection protocol was performed on commercial ham using 2 viral elution buffers (glycine buffer and Tris beef extract buffer) and 2 concentration solutions [polyethylene glycol (PEG) and zirconium hydroxide]. The different approaches were verified by RT-PCR and nested PCR. This method was performed on ham in less than 8 hr by artificial inoculation of serial dilutions of the virus ranging from 1,000 to 1 RT-PCR unit/mL. The viral extraction and concentration method had 10-fold higher sensitivity using the combination of Tris beef extract buffer and PEG as compared to glycine buffer and zirconium hydroxide. This method proved that RT-PCR and nested PCR have the sensitive ability to detect norovirus in commercial ham, in that norovirus was successfully detected in artificially contaminated samples at a detection level as low as 1-10 RT-PCR unit/mL. Overall, such a detection limit suggests this protocol is both quick and efficient in terms of its potential use for detecting norovirus in meat products.

• Journal of Microbiology and Biotechnology
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• v.31 no.9
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• pp.1323-1329
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• 2021

Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a double-stranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

• Journal of Microbiology and Biotechnology
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• v.33 no.7
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• pp.964-972
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• 2023

Bacteriophage endolysins are peptidoglycan hydrolases composed of cell binding domain (CBD) and an enzymatically active domain. A phage endolysin CBD can be used for detecting bacteria owing to its high specificity and sensitivity toward the bacterial cell wall. We aimed to develop a method for detection of Enterococcus faecalis using an endolysin CBD. The gene encoding the CBD of ECP3 phage endolysin was cloned into the Escherichia coli expression vector pET21a. A recombinant protein with a C-terminal 6-His-tag (CBD) was expressed and purified using a His-trap column. CBD was adsorbed onto epoxy magnetic beads (eMBs). The bacterial species specificity and sensitivity of bacterial binding to CBD-eMB complexes were determined using the bacterial colony counting from the magnetic separations after the binding reaction between bacteria and CBD-eMB complexes. E. faecalis could bind to CBD-eMB complexes, but other bacteria (such as Enterococcus faecium, Staphylococcus aureus, Escherichia coli, Acinetobacter baumannii, Streptococcus mutans, and Porphyromonas gingivalis) could not. E. faecalis cells were fixed onto CBD-eMB complexes within 1 h, and >78% of viable E. faecalis cells were recovered. The E. faecalis recovery ratio was not affected by the other bacterial species. The detection limit of the CBD-eMB complex for E. faecalis was >17 CFU/ml. We developed a simple method for the specific detection of E. faecalis using bacteriophage endolysin CBD and MBs. This is the first study to determine that the C-terminal region of ECP3 phage endolysin is a highly specific binding site for E. faecalis among other bacterial species.

• Proceedings of the Korean Magnestics Society Conference
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• 2003.12a
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• pp.72-72
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• 2003