• Title/Summary/Keyword: Affinity techniques

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A study on the application of mill scale-derived magnetite particles for adsorptive removal of phosphate from wastewater (인제거용 흡착제로서 밀스케일로부터 선별된 마그네타이트 적용 연구)

  • Kim, Yunjung;Doliente, Jonica Ella;Choi, Younggyun
    • Journal of Korean Society of Water and Wastewater
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    • v.31 no.4
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    • pp.281-287
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    • 2017
  • Mill scale, an iron waste, was used to separate magnetite particles for the adsorption of phosphate from aqueous solution. Mill scale has a layered structure composed of wustite (FeO), magnetite ($Fe_3O_4$), and hematite ($Fe_2O_3$). Because magnetite shows the highest magnetic property among these iron oxides, it can be easily separated from the crushed mill scale particles. Several techniques were employed to characterize the separated particles. Mill scale-derived magnetite particles exhibited a strong uptake affinity to phosphate in a wide pH range of 3-7, with the maximum adsorptive removal of 100%, at the dosage of 1 g/L, pH 3-5. Langmuir isotherm model well described the equilibrium data, exhibiting maximum adsorption capacities for phosphate up to 4.95 and 8.79 mg/g at 298 and 308 K, respectively. From continuous operation of the packed-bed column reactor operated with different EBCT (empty bed contact time) and adsorbent particle size, the breakthrough of phosphate started after 8-22 days of operation. After regeneration of the column reactor with 0.1N NaOH solution, 95-98% of adsorbed phosphate could be detached from the column reactor.

Expression and Purification of a Cathelicidin-Derived Antimicrobial Peptide, CRAMP

  • Park Eu-Jin;Chae Young-Kee;Lee Jee-Young;Lee Byoung-Jae;Kim Yang-Mee
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1429-1433
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    • 2006
  • Application of recombinant protein production and particularly their isotopic enrichment has stimulated development of a range of novel multidimensional heteronuclear NMR techniques. Peptides in most cases are amenable to assignment and structure determination without the need for isotopic labeling. However, there are many cases where the availability of $^{15}N$ and/or $^{13}C$ labeled peptides is useful to study the structure of peptides with more than 30 residues and the interaction between peptides and membrane. CRAMP (Cathelicidin-Related AntiMicrobial Peptide) was identified from a cDNA clone derived from mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptides. CRAMP was successfully expressed as a GST-fused form in E. coli and purified using affinity chromatography and reverse-phase chromatography. The yield of the CRAMP was 1.5 mg/l 1. According to CD spectra, CRAMP adopted ${\alpha}$-helical conformation in membrane-mimetic environments. Isotope labeling of CRAMP is expected to make it possible to study the structure and dynamic properties of CRAMP in various membrane systems.

A Study on Properties of OLEDs using $Zn(HPB)_2$ as hole blocking layer ($Zn(HPB)_2$를 Hole blocking layer로 이용한 OLEDs의 특성 연구)

  • Kim, Dong-Eun;Kim, Byoung-Sang;Kwon, Oh-Kwan;Lee, Burm-Jong;Kwon, Young-Soo
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2005.07a
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    • pp.447-448
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    • 2005
  • Recently, organic light emitting diodes(OLEDs) is widely used as one of the information display techniques. We synthesized 2-(2-hydroxyphenyl)benzoxazole($Zn(HPB)_2$). We studied the luminescent properties of OLEDs using $Zn(HPB)_2$. The ionization potential(IP) and the electron affinity(EA) of $Zn(HPB)_2$ investigated using cyclic-voltammetry(C-V). The JP, EA and Eg were 6.5eV, 3.0eV and 3.5eV, respectively. The PL and EL spectra of $Zn(HPB)_2$ were observed at the wavelength of 4S0nm. We used $Zn(HPB)_2$ as an emitting layer and hole blocking layer. At the experiment about hole blocking effect, we inserted $Zn(HPB)_2$ between emitting material layer(EML) and cathode, and hole transport layer(HTL) and emitting material layer(EML). We measured current density-voltage and luminance-voltage characteristics at room temperature.

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Radioimmunotherapy (II): Clinical Application (방사면역치료(II): 임상적 이용)

  • Cheon, Gi-Jeong;Kang, Hye-Jin;Lim, Sang-Moo
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.2
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    • pp.74-81
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    • 2006
  • Molecular targeting may be defined as the specific concentration of a diagnostic or therapeutic tracer by its Interaction with a molecular species that is distinctly present or absent in a disease state. Monoclonal antibody (mAb) is one of the successful agents for targeted therapy in cancer. To enhance the therapeutic effect, the concept of targeting radionuclides to tumors using radiolabeled mAbs against tumor-associated antigens, radioimmunotherapy, was proposed. The efficacy of radioimmunotherapy, however, has to be further optimized. Several strategies to improve targeting of tumors with radiolabeled mAbs have been developed, such as the use of mAb fragments, the use of high-affinity mAbs, the use of labeling techniques that are stable in vivo, active removal of the radiolabeled mAb from the circulation, and pretargeting strategies. Until now, however, there are many kinds of obstacles to be solved in the use of mAb for the targeted therapy. Major technical challenges to molecular targeting are related to the rapid and specific delivery of tracers to the target, the elimination of unwanted background activity, and the development of more specific targets to create a cytocidal effect. further development of this field will be determined by success in solving these challenges.

N-terminal formylmethionine as a novel initiator and N-degron of eukaryotic proteins

  • Kim, Jeong-Mok
    • BMB Reports
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    • v.52 no.3
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    • pp.163-164
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    • 2019
  • The ribosomal synthesis of proteins in the eukaryotic cytosol has always been thought to start from the unformylated N-terminal (Nt) methionine (Met). In contrast, in virtually all nascent proteins in bacteria and eukaryotic organelles, such as mitochondria and chloroplasts, Nt-formyl-methionine (fMet) is the first building block of ribosomal synthesis. Through extensive approaches, including mass spectrometric analyses of the N-termini of proteins and molecular genetic techniques with an affinity-purified antibody for Nt-formylation, we investigated whether Nt-formylated proteins could also be produced and have their own metabolic fate in the cytosol of a eukaryote, such as yeast Saccharomyces cerevisiae. We discovered that Nt-formylated proteins could be generated in the cytosol by yeast mitochondrial formyltransferase (Fmt1). These Nt-formylated proteins were massively upregulated in the stationary phase or upon starvation for specific amino acids and were crucial for the adaptation to specific stresses. The stress-activated kinase Gcn2 was strictly required for the upregulation of Nt-formylated proteins by regulating the activity of Fmt1 and its retention in the cytosol. We also found that the Nt-fMet residues of Nt-formylated proteins could be distinct N-terminal degradation signals, termed fMet/N-degrons, and that Psh1 E3 ubiquitin ligase mediated the selective destruction of Nt-formylated proteins as the recognition component of a novel eukaryotic fMet/N-end rule pathway, termed fMet/N-recognin.

Comparative study of Pb (II) adsorption from water on used cardboard and powdered activated carbon

  • Benhafsa, Fouad. Mekhalef;Bouchama, Abdelghani.;Chadli, Aicha.;Tadjer, Belgacem.;Addad, Djelloul.
    • Membrane and Water Treatment
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    • v.13 no.2
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    • pp.73-83
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    • 2022
  • In the present study, we compared the adsorption capacity of Pb (II) from contaminated water of used cardboard (UC) and a commercial powdered activated carbon (PAC), the latter has been characterized by different techniques, namely X-ray diffraction (XRD), scanning electron microscopy with energy dispersive spectroscopy (SEM/EDS), wavelength dispersion x-ray fluorescence (WDXRF), infrared spectroscopy (IR) and surface area B.E.T analyzer. The effect of various parameters, such as the pH, the contact time, the amount of adsorbent, and the temperature on the adsorption of Pb (II) on both materials was investigated. The Pb (II) adsorptions are perfectly described by a pseudo-second-order model, while the intraparticle diffusion is a decisive step after the first minutes of contact. The fit to the Langmuir and Redlich-Peterson models seems perfect for these adsorption reactions. (PAC) showed a greater affinity for Pb (II) compared to (UC) and the adsorption of Pb (II) ions is strongly pH-dependent, on the other hand, the increase in temperature doesn't have much influence on the two solids. This study showed that the capacity of (UC) to adsorb Pb (II) from an aqueous solution is greater than two-thirds of that of (PAC).

Purification and Characterization of Polyphenol Oxidase from Oyster Mushroom (Pleurotus ostreatus) (느타리버섯(Pleurotus ostreatus)의 Polyphenol Oxidase 분리 정제 및 특성 조사)

  • Choi, Ju-Hee;Kim, Hyun-Jin;Park, Sun-Young;Ham, Kyung-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1447-1452
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    • 2011
  • Polyphenol oxidase (PPO) isoforms were partially purified from oyster mushroom (Pleurotus ostreatus) using various chromatography techniques, and their characteristics of heat stability, substrate affinity, optimum pH, and optimum temperature were investigated. Three PPO isoforms named PO-I, PO-II-1, and PO-II-2 were partially purified from oyster mushroom. The molecular weight of PO-II-1 was 70 kDa and PO-I and PO-II-2 were less than 6 kDa each. Characterization was carried out using a PPO isoform partially purified by hydrophobic interaction chromatography. Optimum temperature was $55^{\circ}C$ and optimum pH 5.0. However, the PPO was inactivated at neutral pH or by heating at $80^{\circ}C$ for 30 min, while the 40% PPO still remained active after heating at $60^{\circ}C$ for 45 min. The PPO isoform showed the highest substrate affinity to chlorogenic acid and pyrogallol, in which KM values were 1.01 and 2.06 mM, respectively. Therefore, these results suggested that the mushrooms should be stored at a pH higher than 7.0 and at a low temperature to prevent enzymatic browning.

An Extraction and ANP Importance Analysis for Competence Factors of Junior Grade CM Engineer (초급 건설사업관리기술자(CMr)의 요구역량 추출 및 ANP 중요도 분석)

  • Yang, Jinkook;Kim, Sebeom;Lee, Sangbeom
    • Korean Journal of Construction Engineering and Management
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    • v.18 no.1
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    • pp.58-64
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    • 2017
  • Construction Management (CM) is being utilized as an efficient project management system. Construction management system provide scientific management techniques as well as the expert's service with the expertise. Accordingly, CM services demand is continuously increasing in construction project. Construction Management engineer grade is distinguished by the Index of Construction Engineer's Competency (ICEC). Construction management engineer requires a higher score than the other fields. However, junior grade CM engineer can get grade according to the education and qualification. This study was conducted junior grade CM engineer depth interview after analyzing the problem of the current CM engineer grade system. As a result, this study was to extracted the 23 kind competence factors. And, competence factors were five area grouped using affinity diagram method. Next, we analyzed the importance utilizing the Analytic Network Process (ANP) technique. As a result, high priority areas were analyzed to be quality and cost areas associated with project management.

Receptor Binding Affinities of Synthetic Cannabinoids Determined by Non-Isotopic Receptor Binding Assay

  • Cha, Hye Jin;Song, Yun Jeong;Lee, Da Eun;Kim, Young-Hoon;Shin, Jisoon;Jang, Choon-Gon;Suh, Soo Kyung;Kim, Sung Jin;Yun, Jaesuk
    • Toxicological Research
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    • v.35 no.1
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    • pp.37-44
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    • 2019
  • A major predictor of the efficacy of natural or synthetic cannabinoids is their binding affinity to the cannabinoid type I receptor ($CB_1$) in the central nervous system, as the main psychological effects of cannabinoids are achieved via binding to this receptor. Conventionally, receptor binding assays have been performed using isotopes, which are inconvenient owing to the effects of radioactivity. In the present study, the binding affinities of five cannabinoids for purified $CB_1$ were measured using a surface plasmon resonance (SPR) technique as a putative non-isotopic receptor binding assay. Results were compared with those of a radio-isotope-labeled receptor binding assay. The representative natural cannabinoid ${\Delta}^9$-tetrahydrocannabinol and four synthetic cannabinoids, JWH-015, JWH-210, RCS-4, and JWH-250, were assessed using both the SPR biosensor assay and the conventional isotopic receptor binding assay. The binding affinities of the test substances to $CB_1$ were determined to be (from highest to lowest) $9.52{\times}10^{-3}M$ (JWH-210), $6.54{\times}10^{-12}M$ (JWH-250), $1.56{\times}10^{-11}M$ (${\Delta}^9$-tetrahydrocannabinol), $2.75{\times}10^{-11}M$ (RCS-4), and $6.80{\times}10^{-11}M$ (JWH-015) using the non-isotopic method. Using the conventional isotopic receptor binding assay, the same order of affinities was observed. In conclusion, our results support the use of kinetic analysis via SPR in place of the isotopic receptor binding assay. To replace the receptor binding affinity assay with SPR techniques in routine assays, further studies for method validation will be needed in the future.

Alternative Immunossays

  • Barnard, G.J.R.;Kim, J.B.;Collins, W.P.
    • Korean Journal of Animal Reproduction
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    • v.9 no.2
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    • pp.133-139
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    • 1985
  • An immunoassay may be defined as an analytical procedure involving the competitive reaction between a limiting concentration of specific antibody and two populations of antigen, one of which is labelled or immobillized. The advent of immunoassay has revolutionised our knowledge of reproductive physiology and the practice of veterinary and clinical medicine. Radioimmunoassay (RIA) was the first of these methods to be developed, which meausred the analyte with good sensitivity, accuracy and precision (1,2). The essential components of RIA are:-(i) a limited concentration of antibodies, (ii) a reference preparation, and (iii) an antigen labelled with a radioisotope (usually tritium or iodine-125). Most procedures invelove isolating the antibody-bound fraction and measuring the amount of labelled antigen. Good facilities are available for scintilltion counting, data reduction nd statistical analysis. RIA is undergoing refinement through:-(i) the introduction of new techniques to separate the antibody-bound and free fractions which minimize the misclassification of labelled antigen into these compartments, and the amount of non-specfic binding. (3), (ii) the development of non-extration for the measurement of haptens (4), (iii) the determination of a, pp.rent free (i.e. non-protein bound) analytes (5), and (iv) the use of monoclonal antibodies(6). In 1968, Miles and Hales introduced in important new type of immunoassay which they termed immunora-diometric assay (IRMA) based on t도 use of isotopically labelled specific antibodies(7) in a move from limited to excess reagent systems. The concept of two-site IRMAs (with a capture antibody on a solid-phase, and a second labelled antibody to a different antigenic determinant of the analyte) has enabled the development of more sensitive and less-time consuming methods for the measurement of protein hormones ovar wide concentration of analyte (8). The increasing use of isotopic methos for diverse a, pp.ications has exposed several problems. For example, the radioactive half-life and radiolysis of the labelled reagent limits assay sensitivity and imposes a time limit on the usefulness of a kit. In addition, the potential health hazards associated with the use and disposal of radioactive cmpounds and the solvents and photofluors necessary for liquid scientillation counting are incompatable with the development of extra-laboratory tests. To date, the most practical alternative labels to radioisotopes, for the measurement of analytes in a concentration > 1 ng/ml, are erythrocytes, polystyrene particiles, gold sols, dyes and enzymes or cofactors with a visual or colorimetric end-point(9). Increased sensitivity to<1 pg/ml may be obtained with fluorescent and chemiluminescent labels, or enzymes with a fluorometric, chemiluminometric or bioluminometric end-point. The sensitivity of any immunoassay or immunometric assay depends on the affinity of the antibody-antigen reaction, the specific activity of the label, the precision with which the reagents are manipulated and the nonspecific background signal (10). The sensitivity of a limited reagent system for the measurement of haptens or proteins is mainly dependent upon the affinity of the antibodies and the smalleest amount of reagent that may be manipulated. Consequently, it is difficult in practice to improve on the sensitivity obtained with iodine-125 as the label. Conversely, with excess reagent systems for the measurement of proteins it is theoretically possible to increase assay sensitivity at least 1000 fold with alternative luminescent labels. To date, a 10-fold improvement has been achieved, and attempts are being made to reduce the influence of other variables on the specific signal from the immunoreaction.

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