• 제목/요약/키워드: binding method

검색결과 1,087건 처리시간 0.027초

Detection and Kinetics of Mucosal Pathogenic Bacteria Binding with Polysaccharides

  • Chung, Kyong-Hwan;Park, Jung-Soon;Hwang, Hyun-Soo;Kim, Jin-Chul;Lee, Ki-Young
    • Journal of Microbiology and Biotechnology
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    • 제17권7호
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    • pp.1191-1197
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    • 2007
  • The detection and kinetics of mucosal pathogenic bacteria binding on polysaccharide ligands were studied using a surface plasmon resonance biosensor. The kinetic model applied curve-fitting to the experimental surface plasmon resonance sensorgrams to evaluate the binding interactions. The kinetic parameters for the mucosal pathogenic bacteria (Pseudomonas aeruginosa, Pseudomonas fluorescens, Serratia marcescens) with the alginate ligand were determined from a kinetic model. In addition, the binding interactions of the mucosal pathogenic bacteria with polysaccharide binding pairs (Pseudomonas aeruginosa/alginate, Streptococcus pneumoniae/pneumococcal polysaccharide, Staphylococcus aureus/pectin) were also compared with their kinetic parameters. The rate constants of association for Pseudomonas aeruginosa with the alginate ligand were higher than those for Pseudomonas fluorescens. Serratia marcescens had no detectable interaction with the alginate ligand. The adhesion affinity of Pseudomonas aeruginosa with alginate was higher than that for the other binding pairs. The binding affinities of the pathogenic bacteria with their own polysaccharide were higher than that of Staphylococcus aureus with pectin. Measuring the contact angle was found to be a feasible method for detecting binding interactions between analytes and ligands.

Identification and Characterization of a Novel Angiostatin-binding Protein by the Display Cloning Method

  • Kang, Ha-Tan;Bang, Won-Ki;Yu, Yeon-Gyu
    • BMB Reports
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    • 제37권2호
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    • pp.159-166
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    • 2004
  • Angiostatin is a potent anti-angiogenic protein. To examine the angiostatin-interacting proteins, we used the display-cloning method with a T7 phage library presenting human cDNAs. The specific T7 phage clone that bound to the immobilized angiostatin was isolated, and a novel gene encoding the displayed polypeptide on the isolated T7 phage was identified. The displayed angiostatin-binding sequence was expressed in E. coli as a soluble protein and purified to homogeneity. This novel angiostatin-binding region interacted specifically to angiostatin with a dissociation constant of $3.4{\times}10^{-7}\;M$. A sequence analysis showed that the identified sequence was a part of the large ORF of 1,998 amino acids, whose function has not yet been characterized. A Northern analysis indicated that the gene containing the angiostatin-binding sequence was expressed differentially in the developmental stages or cell types.

접합자 난관내 이식 환자에 있어서 수정 실패와 항정자 항체와의 관계 및 난자와 정자의 재처리에 관한 연구 (Study on the Detection of Anti-Sperm Antibodies in Zygote Intra Fallopian Transfer (ZIFT) Patients with Fertilization Failure or Low Fertilization Rate and Retreatment of Oocyte and Sperm)

  • 정미경;고정재;도병록;구정진;한세열;차광열
    • Clinical and Experimental Reproductive Medicine
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    • 제19권2호
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    • pp.169-174
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    • 1992
  • Previous studies have indicated that immunological factor is responsible for the infertility. We have detected sperm antibodies in ZIFT patients which grouped as fertilization failure (A; n=18) and low fertilization rate (${\leq}50%$)(B; n=20). Patients, however, had normal oocytes and sperms. We collected serum from wives and semen from husbands and donors (fertile sperm), if it was needed. We examined class, binding patterns and amounts of antisperm antibodies(ASA) by direct and indirect immunobead binding assay. In group A, 11 husbands were ASA positive showing 62.2% and 61.1% binding with IgA and IgG, respectively, and two wives were ASA positive showing 70.0% and 71.0% binding with IgA and IgG, respectively. Binding sites were mainly at the head of sperms (84%). In group B, 8 husbands were ASA positive showing 37.5% and 40.0% binding with IgA and IgG, respectively, and two wives were ASA positive showing 41.3% and 42.0% binding with IgA and IgG, respectively. Binding sites were also mainly at the head of sperms (78%). For the treatment of ZIFT patients who had fertilization failure at the first trial, we used albumin fractionation method and dilution method with 30% fetal cord serum (FCS) to reduce the titer of ASA. We used partial zona dissection (P.Z.D.) method for wives who have antisperm antibodies in their serum. According to represented method, we could inhance the fertilization rate to 60.0% by albumin fractionation and 20.0% by P.Z.D., respectively. We concluded that the use of micromanipulation like P.Z.D. or the other sperm processing methods is required to increase a chance of fertilization. This result suggested that it should be a prerequisite to test antisperm antibodies prior to entering assisted reproductive technologies (ART) programs.

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약물과 생체고분자 간의 상호작용(제 13보)-세파로친과 소혈청알부민의 결합에 미치는 이온강도, pH 및 온도의 영향 (Drug-Biomacromolecule Interaction (XIII)-Effect of ionic Strength, pH and Temperature on Binding of Cephalothin to Bovine Serum Albumin-)

  • 김종국;임연수;양지선;정은주
    • Journal of Pharmaceutical Investigation
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    • 제19권3호
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    • pp.163-171
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    • 1989
  • To investigate the protein binding characteristics of cephalothin, the effects of ionic strength, pH and temperature on the binding of cephalothin to bovine serum albumin (BSA) were studied by UV difference spectrophotometric method. With increasing ionic strength at constant PH and temperature, association constant decreased, but the number of binding sites sites was about 2 constantly. It may be deduced that the binding process is not only due to electrostatic forces. And the increased association constant at high ionic strength is explained by conformational changes of BSA from complex to subunits. The pH effect on the affinity of interaction indicated that the binding affinity of drug is higher in the neutral region than in the alkaline region. And, at high pH value, the number of binding sites decreased from 2 to 1 because of the conformational changes of BSA in alkaline region. The decrease in binding affinity of BSA to drug with increasing temperature was characteristic of an exothermic reaction. And the negative sign of ${\Delta}G^{\circ}$ meant that the binding process occurs spontaneously under the experimental conditions. In cephalothin-BSA complex formation, since the net enthalpy change value and entropy change value are positive, it is assumed that hydrophobic bindings are predominant in this binding process.

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가중치법을 이용한 농작물 지지대 및 결속장치의 최적설계 (Optimum Design of the Agricultural Support and Binder for Stretching Device)

  • 이만기;김진호;신기열
    • 한국기계가공학회지
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    • 제14권4호
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    • pp.28-33
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    • 2015
  • In this study, the optimal design for the support and the binding device for the protection of crops for the maximum allowable stress of the shape necessary to minimize volume has been proposed. Optimization of the support and the binding device for the crops should be designed to support businesses in terms of profit, in part to reduce the material, and to profit from the ease and speed of working that part of the farmers. We used CATIA for the mechanical design and the ANSYS program for the structural analysis. Additionally, the optimization was performed by PIAnO with seven design variables for the binding device and three parameters for the support. The weight method using a multi-objective function was also determined by the Pareto optimal solution. The volume of the binding device in the optimum design result was found to be reduced by 16%, from $2.278e-005m^3to1.912e-005m^3$. From the result, we confirmed the effectiveness of the design method proposed as a multi-objective function optimization problem.

Genetic effects of sterol regulatory element binding proteins and fatty acid-binding protein4 on the fatty acid composition of Korean cattle (Hanwoo)

  • Oh, Dong-Yep;Lee, Jea-Young;Jang, Ji-Eun;Lee, Seung-Uk
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권2호
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    • pp.160-166
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    • 2017
  • Objective: This study identifies single-nucleotide polymorphisms (SNP) or gene combinations that affect the flavor and quality of Korean cattle (Hanwoo) by using the SNP Harvester method. Methods: Four economic traits (oleic acid [C18:1], saturated fatty acids), monounsaturated fatty acids, and marbling score) were adjusted for environmental factors in order to focus solely on genetic effects. The SNP Harvester method was used to investigate gene combinations (two-way gene interactions) associated with these economic traits. Further, a multifactor dimensionality reduction method was used to identify superior genotypes in gene combinations. Results: Table 3 to 4 show the analysis results for differences between superior genotypes and others for selected major gene combinations using the multifactor dimensionality reduction method. Environmental factors were adjusted for in order to evaluate only the genetic effect. Table 5 shows the adjustment effect by comparing the accuracy before and after correction in two-way gene interactions. Conclusion: The g.3977-325 T>C and (g.2988 A>G, g.3977-325 T>C) combinations of fatty acid-binding protein4 were the superior gene, and the superior genotype combinations across all economic traits were the CC genotype at g.3977-325 T>C and the AACC, GACC, GGCC genotypes of (g.2988 A>G, g.3977-325 T>C).

냉동절편을 이용한 돼지 자궁내 에스트로겐 수용체의 측정 (A Method for the Determination of Estrogen Receptor Level in Frozen Sections of Porcine Uterus)

  • 윤용달;박철홍;이영근
    • Clinical and Experimental Reproductive Medicine
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    • 제16권2호
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    • pp.131-138
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    • 1989
  • The present study was designed to develop a new method for the determination of estrogen receptor in porcine uterus using frozen sections. Cryostat sections were incubated with $^3H$-estradiol($^3H$-$E_2$) in the presence or absence of diethylstilbestrol(DES) and the radioactivity of 3H-E2 bound to estrogen receptor(ER) was detected. The level of specific estrogen receptor was determined by Scatchard analysis. The highest ratio of specific binding against total binding was achieved in 3 sec. tions(5mm x 5mm) which was corresponded to lOO${\mu}$/ml protein concentration. Optimal binding was obtained during incubation with $^3H$-$E_2$ for 30 minutes at 23$^{\circ}C$ after treatment of sections with acetone for 20 seconds. Three time-washing of sections was proved to be appropriate for the removal of unbound 3H-E2. 200-fold molar excess of DES was substituted for the binding of $^3H$-$E_2$ to ER sufficiently(binding efficiency of 54.8%). ER was saturated with 4nM of $^3H$-$E_2$ and its dissociation constant was 0.1nM. ER assay using frozen sections(Histological radioreceptor assay, HRRA) was significantly correlated with radioreceptor assay for estradiol(RRA, 0.976 , p

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Prediction of Binding Free Energy Calculation Using Molecular Mechanics/Poisson-Boltzmann Surface Area (MM-PBSA) Method in Drug Discovery: A Short Review

  • Kothandan, Gugan;Cho, Seung Joo
    • 통합자연과학논문집
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    • 제5권4호
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    • pp.216-219
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    • 2012
  • Structure-based drug design possibly benefit from in silico methods that precisely predict the binding affinity of small molecules to target macromolecules. There are many limitations arise from the difficulty of predicting the binding affinity of a small molecule to a biological target with the current scoring functions. There is thus a strong interest in novel methodologies based on MD simulations that claim predictions of greater accuracy than current scoring functions, helpful for a regular use designed for drug discovery in the pharmaceutical industry. Herein, we report a short review on free energy calculations using MMPBSA method a useful method in structure based drug discovery.

Identification of a Bacteria-Specific Binding Protein from the Sequenced Bacterial Genome

  • Kong, Minsuk;Ryu, Sangryeol
    • Journal of Microbiology and Biotechnology
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    • 제26권1호
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    • pp.38-43
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    • 2016
  • Novel and specific recognition elements are of central importance in the development of a pathogen detection method. Here, we describe a simple method for identifying the cell-wall binding domain (CBD) from a sequenced bacterial genome employing homology search for phage lysin genes. A putative CBD (CPF369_CBD) was identified from a genome of Clostridium perfringens type strain ATCC 13124, and its function was studied with the CBD-GFP fusion protein recombinantly expressed in Escherichia coli. Fluorescence microscopy showed the specific binding of the fusion protein to C. perfringens cells, which demonstrates the potential of this method for the identification of novel bioprobes for specific detection of pathogenic bacteria.

뉴톤법에 의한 최적전력 조류계산 (Optimal Power Flow Study by The Newton's Method)

  • 황갑주
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 1989년도 하계종합학술대회 논문집
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    • pp.173-178
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    • 1989
  • Optimal Power Flow (OPF) solution by the Newton's method provides a reliable and robust method to classical OPF problems. The major challenge in algorithm development is to identify the binding inequalities efficiently. This paper propose a simple strategy to identify the binding set. From the mechanism of penalty shifting with soft penalty in trial iteration, a active binding sit is identified automatically. This paper also suggests a technique to solve the linear system whore coefficients are presented by the matrix. This implementation is highly efficient for sparsity programming. Case study for 3,5,14,118,190 bus and practrical KEPCO 305 bus system are performed as well.

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