• 제목/요약/키워드: Affinity techniques

검색결과 65건 처리시간 0.032초

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

  • 김윤중;엘라;최영균
    • 상하수도학회지
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    • 제31권4호
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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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    • 제16권9호
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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.

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

  • 김동은;김병상;권오관;이범종;권영수
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2005년도 하계학술대회 논문집 Vol.6
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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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방사면역치료(II): 임상적 이용 (Radioimmunotherapy (II): Clinical Application)

  • 천기정;강혜진;임상무
    • Nuclear Medicine and Molecular Imaging
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    • 제40권2호
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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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    • 제52권3호
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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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    • 제13권2호
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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).

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

  • 최주희;김현진;박선영;함경식
    • 한국식품영양과학회지
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    • 제40권10호
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    • pp.1447-1452
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    • 2011
  • 우리나라 버섯 생산량의 약 80%를 차지하는 느타리버섯(Pleurotus ostreatus)의 유통 중에 발생하는 갈변을 방지하기 위한 기초 조사로 갈변원인 효소인 polyphenol oxidase(PPO)를 분리 정제하여 그 특성을 조사하였다. 느타리버섯으로부터 3개의 PPO isoforms(PO-I, PO-II-1, PO-II-2)을 분리하였으며, gel-filteration을 이용한 분자량 확인 결과 PO-II-1은 void volume(70 kDa), PO-I과 PO-II-2는 include volume(6 kDa)에서 분리되는 것으로 보아 PO-II-1은 70 kDa 이상, PO-I과 PO-II-2는 6 kDa 이하인 것으로 확인되었다. 분리된 3개의 PPO isoform들을 partially denatured-PAGE 후, activity staining을 이용하여 분석한 결과, 비록 이들 PPO isoforms PO-II-1과 PO-II-2에 다수의 단백질들이 들어있기는 하지만 PPO 활성을 보이는 band가 하나로 나타남으로써 분리된 PPO isoform들이 실제 PPO 활성을 갖고 있는 단백질로 확인되었으며 다른 PPO isoform이 혼합되지 않은 단일 PPO isoform으로 분리되었음을 알 수 있다. Hydrophobic interaction chromatography에서 분리된 isoform PO를 이용하여 특성을 조사한 결과 최적 반응온도와 pH는 일반적인 다른 과채류와는 달리 $50{\sim}55^{\circ}C$, pH 5.5에서 높은 활성을 나타내었으며, 열 안정성 실험에 있어서는 $60^{\circ}C$에서 45분간 가열 처리 시 약 40%의 PPO activity가 남아있는 반면, $80^{\circ}C$에서 30분간 가열 처리 시 PPO가 완전히 불 활성화되었다. 그리고 chlorogenic acid와 pyrogallol에 대하여 높은 기질 특이성을 보였다.

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

  • 양진국;김세범;이상범
    • 한국건설관리학회논문집
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    • 제18권1호
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    • pp.58-64
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    • 2017
  • 건설관리 (CM)는 효율적인 프로젝트 관리 시스템으로서 활용되고 있다. 건설관리 시스템은 과학적 관리 기법을 제공할 뿐만 아니라, 전문 지식을 가진 전문가의 서비스를 제공한다. 따라서 CM 서비스 수요는 건설 프로젝트에서 지속적으로 증가하고 있다. 건설사업관리기술자의 등급은 건설 기술자 역량지수 (ICEC)에 의해 구분된다. 건설사업관리기술자는 다른 분야보다 높은 점수를 필요로 한다. 하지만 초급 건설사업관리기술자는 교육과 자격에 따라 등급을 획득할 수 있다. 본 연구는 현행 건설사업관리기술자 등급 시스템의 문제점을 분석한 후 초급 건설사업관리기술자를 대상으로 인터뷰를 실시하였다. 그 결과 본 연구에서는 23가지의 요구역량 요인들을 추출하였다. 그리고 요구역량은 친화도법을 사용하여 5가지 영역으로 그룹화 하였다. 다음으로, ANP 기법을 이용하여 중요도를 분석하였다. 그 결과 우선 순위가 높은 영역이 프로젝트 관리와 관련된 품질 및 비용 영역으로 분석되었다.

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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    • 제35권1호
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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.
    • 한국가축번식학회지
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    • 제9권2호
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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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