• Environmental Engineering Research
• /
• v.14 no.1
• /
• pp.63-67
• /
• 2009

Escherichia coli K-12 (E. coli K-12) is a representative indicator globally used for distinguishing and monitoring dynamic fates of pathogenic microorganisms in the environment. This study investigated how to most critically quantify lacZ ($\beta$-galactosidase) gene in E. coli K-12 by two different real-time polymerase chain reaction (real-time PCR) in association with three different DNA extraction practices. Three DNA extractions, i.e., sodium dodecyl sulfate (SDS)/proteinase K, magnetic beads and guanidium thiocyanate (GTC)/silica matrix were each compared for extracting total genomic DNA from E. coli K-12. Among them, GTC/silica matrix and magnetic beads beating similarly worked out to have the highest (22-23 ng/${\mu}L$) concentration of DNA extracted, but employing SDS/proteinase K had the lowest (10 ng/${\mu}L$) concentration of DNA retrieved. There were no significant differences in the quantification of the copy numbers of lacZ gene between SYBR Green I qPCR and QProbe-qPCR. However, SYBR Green I qPCR obtained somewhat higher copy number as $1{\times}10^8$ copies. It was decided that GTC/silica matrix extraction or magnetic beads beating in combination with SYBR Green I qPCR can be preferably applied for more effectively quantifying specific gene from a pure culture of microorganism.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.5
• /
• pp.1911-1915
• /
• 2012

Aim: New technologies for the early detection of pancreatic cancer (PC) are urgently needed. The aim of the present study was to screen for the potential protein biomarkers in serum using proteomic fingerprint technology. Methods: Magnetic beads combined with surface-enhanced laser desorption/ionization (SELDI) TOF MS were used to profile and compare the protein spectra of serum samples from 85 patients with pancreatic cancer, 50 patients with acute-on-chronic pancreatitis and 98 healthy blood donors. Proteomic patterns associated with pancreatic cancer were identified with Biomarker Patterns Software. Results: A total of 37 differential m/z peaks were identified that were related to PC (P < 0.01). A tree model of biomarkers was constructed with the software based on the three biomarkers (7762 Da, 8560 Da, 11654 Da), this showing excellent separation between pancreatic cancer and non-cancer., with a sensitivity of 93.3% and a specificity of 95.6%. Blind test data showed a sensitivity of 88% and a specificity of 91.4%. Conclusions: The results suggested that serum biomarkers for pancreatic cancer can be detected using SELDI-TOF-MS combined with magnetic beads. Application of combined biomarkers may provide a powerful and reliable diagnostic method for pancreatic cancer with a high sensitivity and specificity.

• Proceedings of the Korean Magnestics Society Conference
• /
• 2008.12a
• /
• pp.147.2-147.2
• /
• 2008

• Journal of the Korean Magnetics Society
• /
• v.24 no.4
• /
• pp.101-106
• /
• 2014

The glass/Ta(5.8 nm)/NiFe(5 nm)/Cu(2.3 nm)/NiFe(3 nm)/IrMn(12 nm)/Ta(5.8 nm) GMR-SV (giantmagneto-resistance-spin valve) multilayer structure films with a magnetoresistance ratio (MR) of 5.0 % and a magnetic sensitivity (MS) of 1.5%/Oe was deposited by dc magnetron sputtering method. Also, GMR-SV device having a width of $7{\mu}m{\sim}8{\mu}m$ similar to the diameter of RBC (red blood cell) was fabricated by the light lithography process. When RBCs coupled with several magnetic beads with a diameter of $1{\mu}m$ dropped upon the GMR-SV device having MR = 1.06% and MS = 0.3 %/Oe, there is observed the variation of about included of a resistance value of ${\Delta}R=0.4{\Omega}$ and ${\Delta}MR=0.15%$ around a external magnetic field of -0.6 Oe. 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 of hemoglobin inside of RBC combined to magnetic beads.

• Journal of Biomedical Engineering Research
• /
• v.28 no.1
• /
• pp.102-107
• /
• 2007

In molecular imaging studies via magnetic resonance imaging, in vivo cell tracking is an important issue for the observation of cell therapy or disease behavior. High resolution imaging and longitudinal study are necessary to track the cell movement. Since the field inhomogeneity extends over several voxels, we have performed the numerical analysis using the sub-voxel method dividing a voxel of MR image into several elements and the information about the field inhomogeneity distribution around the micro-beads. We imbedded ferrite-composite micro-beads with the size of $20-150{\mu}m$ in the subject substituted for cells to induce local field distortion. In the phantom imaging with the isotropic voxel size of $200{\mu}m^3$, we could confirm the feasibility of sub-voxel tracking in a 3.0 T MRI.

• Journal of Environmental Health Sciences
• /
• v.18 no.1
• /
• pp.84-94
• /
• 1992

In an attempt to develop the immobilized biocatalysts based on immobilized Saccharomyces cerevisiae, immobilized yeast was investigated with respect to the conditions affected to ethanol productivities. Saccharomyces cerevisiae was immobilized in the form of the beads by magnetic-calcium alginate, non magnetic-calcium alginate and acrylamide polymerization. Magnetic immobilized yeast, nonmagnetic immobilized yeast and polyacrylamide immobilized yeast were compared in respect of their pH stability, thermostability, heat tolerance, the relation between the concetration of native yeast and retained activity of immobilized yeast, the activity depending on bead size of immobilized yeast, and the effects of magnesium and cobalt on the activities. The more small bead had retained the higher activity for the three kinds of immobilized yeast. In case of 1.0mm diameter of beads, the retained activity was 40~50% for the all groups. The pH stability profile for the immobilized yeast showed a broad range of optimun activity while the native yeast gave a sharp pick for its optimun pH value. The thermostability was at the range of 25~55$^{\circ}$C for the immobilized yeast groups. It was investigated that the influent magnesium and cobalt concentration, and the relative activity have an influent on heat tolerance at steady state. Both protein content released from immobilized yeast and activity of immobilized yeast were changed after activation of immobilized yeast cell.

• Transactions of the Korean Society of Mechanical Engineers B
• /
• v.32 no.11
• /
• pp.856-862
• /
• 2008

This paper presents the characterization of a continuous magnetophoretic microseparator for separating white and red blood cells from peripheral whole blood cells based on their native magnetic properties. The magnetophoretic microseparator separated the blood cells using a high gradient magnetic separation (HGMS) method without the use of additives such as magnetic beads or probing materials. Experimental results show that the paramagnetic capture mode microseparator can continuously separate out 93.5% of red blood cells and 97.4% of white blood cells from diluted whole blood, and the diamagnetic capture mode microseparator can continuously separate out 89.7% of red blood cells and 72.7 % of white blood cells by using applying an external magnetic flux of 0.2 T using a permanent magnet.

• Journal of Sensor Science and Technology
• /
• v.15 no.3
• /
• pp.158-163
• /
• 2006

This paper reports a novel immunoassay method using superparamagnetic nanoparticles and an enhanced magnetic field gradient for the detection of protein in a microfluidic device. We use superparamagnetic nanoparticles as a label and fluorescent polystyrene beads as a solid support. Based on this platform, magnetic force-based microfluidic immunoassay is successfully applied to analyze the concentration of IgG as model analytes. In addition, we present ferromagnetic microstructure connected with a permanent magnet to increase magnetic flux density gradient (dB/dx, ${\sim}10^{4}$ T/m), which makes limit of detection reduced. The detection limit is reduced to about 1 pg/mL.

• Proceedings of the Korean Magnestics Society Conference
• /
• 2008.12a
• /
• pp.248.1-248.1
• /
• 2008

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