• KSBB Journal
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• v.30 no.2
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• pp.82-90
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• 2015

Plant cell immobilization can protect plant cells from shear forces and increase the stability of gene. An additional advantage of immobilization is the easiness for performing continuous culture with cell recycling. Therefore plant cell immobilization can overcome the limitations of plant cell applications. In addition, target protein should be selected from pharmaceutical proteins to get rid of low expression level problem. The enhanced production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was investigated in immobilized Nicotiana tabacum suspension cell cultures. When the cells were immobilized in polyurethane foam, specific production of hGM-CSF was higher than that in alginate bead immobilization. Optimum continuous culture condition was the addition of 60 g/L sucrose in growth media with exchanging media every 6 day. Under the same condition, specific hGM-CSF production was 7 times higher in a 500-mL spinner flask than that in 100-mL Erlenmeyer flasks. Therefore, development of an effective immobilization process would be possible when the advantage of easy cell recycling was used. Consequently, enhanced production of target proteins could be possible in immobilized continuous cultures when the advantages of immobilization were applied.

• Proceedings of the Korean Environmental Health Society Conference
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• 2004.12a
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• pp.51-62
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• 2004

Lime based stabilization/solidification (S/S) can be an effective remediation alternative for the immobilization of arsenic (As) in contaminated soils and sludges. However, the exact immobilization mechanism has not been well established, Based on previous research, As immobilization could be attributed to sorption and/or inclusion in pozzolanic reaction products and/or the formation of calcium-arsenic (Ca-As) precipitates. In this study, suspensions of lime-As were studied in an attempt to elucidate the controlling mechanism of As immobilization in lime treated soils. Aqueous lime-As suspensions (slurries) with varying Ca/As molar ratios (1:1, 1.5:1, 2:1, 2.5:1 and 4:1) were prepared and soluble As concentrations were determined. X-ray diffraction (XRD) analyses were used to establish the resulting mineralogy of crystalline precipitate formation. Depending on the redox state of the As source, different As precipitates were identified. When As (III) was used, the main precipitate formation was Ca-As-O. With As(V) as the source, Ca4(OH)2(AsO4)2${\cdot}$4H2O formed at Ca/As molar ratios greater than 1:1. A significant increase in As (III) immobilization was observed at Ca/As molar ratios greater than 1:1. Similarly, a substantial increase in As (V) immobilization was noted at Ca/As molar ratios greater than or equal to 2.5: 1. This observation was also confirmed by XRD. The effectiveness of both As (III) and As(V) immobilization in these slurries appeared to increase with increasing Ca/As molar ratios.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.12
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• pp.5115-5118
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• 2015

Background: The $p16^{INK4a}$ is a protein that expressed in Liquid-based cervical cytology specimens and has been proved link to cervical cancer. The $p16^{INK4a}$ could be detection by piezoelectric immunosensor and the immobilization of the $p16^{INK4a}$ antibody influence the sensitivity of the piezoelectric immunosensor. Materials and Methods: $5{\mu}L$ mouse polyclonal antibody against $p16^{INK4a}$ was bound onto the surface of immonosensor through two methods. (directly immobilized method; protein A method). Absorb of the $p16^{INK4a}$ antibody on the surface of immonosensor caused a shift in the resonant frequency of the immunosensor and The frequency changes recorded showed a better reproducibility. The activity of the immobilization antibody with the directly method and protein A method was tested with $p16^{INK4a}$ antigen. Results: The resonant frequency for different antibody immobilization methods were different, and the sensitivity for $p16^{INK4a}$ detection also different. Conclusions: The protein A method was found to be much more better than the directly method for the immobilization of the p16INK4A antibody on the gold electrode of the quartz crystal for cervical lesion detection. The Protein A method created more reproducible and stable immobilization antibody layers with p16INK4A antigen.

• Journal of Microbiology and Biotechnology
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• v.22 no.5
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• pp.628-636
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• 2012

Raw-starch-digesting enzyme (RSDA) was immobilized on Amberlite beads by conjugation of glutaraldehyde/polyglutaraldehyde (PG)-activated beads or by crosslinking. The effect of immobilization on enzyme stability and catalytic efficiency was evaluated. Immobilization conditions greatly influenced the immobilization efficiency. Optimum pH values shifted from pH 5 to 6 for spontaneous crosslinking and sequential crosslinking, to pH 6-8 for RSDA covalently attached on polyglutaraldehyde-activated Amberlite beads, and to pH 7 for RSDA on glutaraldehyde-activated Amberlite. RSDA on glutaraldehyde-activated Amberlite beads had no loss of activity after 2 h storage at pH 9; enzyme on PG-activated beads lost 9%, whereas soluble enzyme lost 65% of its initial activity. Soluble enzyme lost 50% initial activity after 3 h incubation at $60^{\circ}C$, whereas glutaraldehyde-activated derivative lost only 7.7% initial activity. RSDA derivatives retained over 90% activity after 10 batch reuse at $40^{\circ}C$. The apparent $K_m$ of the enzyme reduced from 0.35 mg/ml to 0.32 mg/ml for RSDA on glutaraldehyde-activated RSDA but increased to 0.42 mg/ml for the PG-activated RSDA derivative. Covalent immobilization on glutaraldehyde Amberlite beads was most stable and promises to address the instability and contamination issues that impede the industrial use of RSDAs. Moreover, the cheap, porous, and non-toxic nature of Amberlite, ease of immobilization, and high yield make it more interesting for the immobilization of this enzyme.

• Journal of Microbiology and Biotechnology
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• v.23 no.2
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• pp.205-210
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• 2013

Thiol-ene polymerization is a versatile tool for several applications. Here we report the preparation of epoxide groups containing thiol-ene photocurable polymeric support and the covalent immobilization of ${\alpha}$-amylase onto these polymeric materials. The morphology of the polymeric support was characterized by scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) coupled with SEM was used to explore the chemical composition. The polymeric support and the immobilization of the enzyme were characterized by FTIR analysis. SEM-EDS and FTIR results showed that the enzyme was successfully covalently attached to the polymeric support. The immobilization efficiency and enzyme activity of ${\alpha}$-amylase were examined at various pH (5.0-8.0) and temperature ($30-80^{\circ}C$) values. The storage stability and reusability of immobilized ${\alpha}$-amylase were investigated. The immobilization yield was $276{\pm}1.6$ mg per gram of polymeric support. Enzyme assays demonstrated that the immobilized enzyme exhibited better thermostability than the free one. The storage stability and reusability were improved by the immobilization on this enzyme support. Free enzyme lost its activity completely within 15 days. On the other hand, the immobilized enzyme retained 86.7% of its activity after 30 days. These results confirm that ${\alpha}$-amylase was successfully immobilized and gained a more stable character compared with the free one.

• Journal of Applied Biological Chemistry
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• v.49 no.1
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• pp.28-32
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• 2006

Effect of organic-C on immobilization and nitrification patterns in acidic soil was examined during 20 weeks incubation period to verify if organic amendments such as composted material can increase soil retention of N by stimulating microbial immobilization of $NH_4^+$. Four treatments were laid out: control without fertilizer N and glucose (treatment code: S), ammonium sulfate (SN), ammonium sulfate with single glucose at the commencement (0 week) of incubation (SNG), and ammonium sulfate with double glucose at 0 and 4 weeks of incubation (SNGG). Glucose application (SNG) significantly increased microbial immobilization of $NH_4^+$ within 1 week of incubation over SN. Immobilization was followed by remineralization thereafter; however, second-application of glucose (SNGG) restored $NH_4^+$ immobilization. At the same time, nitrification was significantly inhibited by glucose application as indicated by consistently low $NO_3^-$ concentration in SNG and SNGG soils, suggesting that microbial assimilation of $NH_4^+$ is predominant compared to nitrification when available C-source is abundant. These results suggest application of chemical fertilizer-N with organic amendment would have beneficial effect on soil-N retention and environmental conservation by reducing production of $NO_3^-$ which is likely to be lost through leaching or denitrification.

• Membrane and Water Treatment
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• v.11 no.3
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• pp.195-200
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• 2020

In this study, a biocatalyst composite was prepared by immobilizing oxidoreductases onto Cu-activated zeolite to facilitate biochemical decomposition of 4-chlorophenol (4-CP). 4-CP monooxygenase (CphC-I) was cloned from a 4-CP degrading bacterium, Pseudarthrobacter chlorophenolicus A6, and then overexpressed and purified. Type X zeolite was synthesized from non-magnetic coal fly ash using acetic acid treatment, and its surfaces were coated with copper ions via impregnation (Cu-zeolite). Then, the recombinant oxidative and reductive enzymes were immobilized onto Cu-zeolite. The enzymes were effectively immobilized onto the Cu-zeolite (79% of immobilization yield). The retained catalytic activity of CphC-I after immobilization was 0.3423 U/g-Cu-zeolite, which was 63.3% of the value of free enzymes. The results of this study suggest that copper can be used as an effective enzyme immobilization binder because it provides favorable metalhistidine binding between the enzyme and Cu-zeolite.

• Journal of the Korean Electrochemical Society
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• v.13 no.1
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• pp.70-74
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• 2010

The immobilization of proteins on silicon surfaces using electrochemical reaction has been studied. Chemical deposition of nitrobenzendiazonium (NiBD) cations is employed to modify silicon surfaces. Electrochemical reduction of nitro-group to primary amine-group have been conducted on the modified surfaces to activate silicon surfaces for the protein immobilization. Attachment of gold nanoparticles was used to prove the reduction. The current method was applied to selective activation of a silicon nanowire and immobilize proteins on the selected nanowire. It has been demonstrated that the use of chemical and electrochemical reaction NiBD is efficient for the selective immobilization of proteins on silicon nanowire surfaces.

• Journal of Oriental Neuropsychiatry
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• v.25 no.2
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• pp.191-202
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• 2014

Objectives: The purpose of this study was to examine the anti-depressive effects of BanHaHuBakTang-kami (BHHBT) on an animal model of depression induced by chronic immobility stress. Methods: Mice were treated daily with immobilization stress for 2 hours over a period of 21 days. To examine the effect of BHHBT, we performed behavioral, biochemical and histological analysis to measure immobility time (FST), brain neurotransmitter concentration (HPLC, ELISA), hippocampal damage (H&E staining) and CRF-R1 expression (immunohistochemistry). Results: BHHBT has reduced the immobility time of immobilization stress exposed mice in the forced swimming test. BHHBT has increased the amount of serotonin in the brain. BHHBT has increased the expression level of serotonin in the brain. BHHBT 540 mg/kg were sufficient to prevent tissue damage in the hippocampus region. BHHBT has reduced the expression level of CRF receptors in the hippocampus region. Conclusions: These results suggest that BHHBT may have anti-depressive effects on mice treated with immobilization stress by reducing immobility, increasing brain serotonin concentration and reducing CRF-R1 expression in the hippocampus region.

• Journal of Physiology & Pathology in Korean Medicine
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• v.28 no.4
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• pp.403-410
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• 2014

The purpose of this study was to examine the anti-depressive effects of OnDam-Tang with addition of Linderae Radix (ODT-L) on the animal model of depression induced by chronic immobilization stress. Depression model was made by chronic immobilization stress for 2 hours for 21 days. And we performed forced swimming test, analysis of the neurotransmitter and immunohistochemical staining, measured expression levels of serotonin in the brain. ODT-L has decreased immobilization time in forced swimming test. ODT-L has increased amount of melatonin in the brain. ODT-L has increased expression levels of serotonin in the brain. ODT-L prevented damage in the hippocampal region. ODT-L has reduced the expression level of CRF receptors in in hippocampus region. These results suggest that ODT-L may have anti-depressive effects on depression.