• 제목/요약/키워드: binding method

검색결과 1,087건 처리시간 0.027초

Quantitative Assay of Hepatitis B Surface Antigen by Using Surface Plasmon Resonance Biosensor

  • Hwang, Sang-Yoon;Yoo, Chang-Hoon;Jeon, Jun-Yeoung;Choi, Sung-Chul;Lee, Eun-Kyu
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권4호
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    • pp.309-314
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    • 2005
  • We performed a basic experiment for the rapid, on-line, real-time measurement of hepatitis B surface antigen using a surface plasmon resonance biosensor. We immobilized anti­HBsAg (hepatitis B surface antigen) polyclonal antibody, as a ligand, to the dextran layer on a CM5 chip surface that had previously been activated by N-hydroxysuccinimide. A sample solution containing HBsAg was fed through a microfluidic channel, and the reflecting angle change due to the mass increase from the binding was detected. The binding characteristics between HBsAg and its polyclonal antibody followed the typical monolayer adsorption isotherm. When the entire immobilized antibody had interacted, no additional, non-specific binding occurred, suggesting the immunoreaction was very specific. The bound antigen per unit mass of the antibody was independent of the immobilized ligand density. No significant steric hindrance was observed at an immobilization density of approximately $17.6 ng/mm^2$. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the ligand was linear up to ca. $40{\mu}g$/mL. This linearity was much higher than that of the ELISA method. It appeared the anti­gen-antibody binding increased as the immobilized ligand density increased. In summary, this study showed the potential of this SPR biosensor-based method as a rapid, simple and multi­sample on-line assay. Once properly validated, it may serve as a more efficient method for HBsAg quantification for replacing the ELISA.

Development of a Magnetic Bead-Based Method for Specific Detection of Enterococcus faecalis Using C-Terminal Domain of ECP3 Phage Endolysin

  • Yoon-Jung Choi;Shukho Kim;Jungmin Kim
    • Journal of Microbiology and Biotechnology
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    • 제33권7호
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    • pp.964-972
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    • 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.

Mesoscale simulation of chloride diffusion in concrete considering the binding capacity and concentration dependence

  • Wang, Licheng;Ueda, Tamon
    • Computers and Concrete
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    • 제8권2호
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    • pp.125-142
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    • 2011
  • In the present paper, a numerical simulation method based on mesoscopic composite structure of concrete, the truss network model, is developed to evaluate the diffusivity of concrete in order to account for the microstructure of concrete, the binding effect of chloride ions and the chloride concentration dependence. In the model, concrete is described as a three-phase composite, consisting of mortar, coarse aggregates and the interfacial transition zones (ITZs) between them. The advantage of the current model is that it can easily represent the movement of mass (e.g. water or chloride ions) through ITZs or the potential cracks within concrete. An analytical method to estimate the chloride diffusivity of mortar and ITZ, which are both treated as homogenious materials in the model, is introduced in terms of water-to-cement ratio (w/c) and sand volume fraction. Using the newly developed approaches, the effect of cracking of concrete on chloride diffusion is reflected by means of the similar process as that in the test. The results of calculation give close match with experimental observations. Furthermore, with consideration of the binding capacity of chloride ions to cement paste and the concentration dependence for diffusivity, the one-dimensional nonlinear diffusion equation is established, as well as its finite difference form in terms of the truss network model. A series of numerical analysises performed on the model find that the chloride diffusion is substantially influenced by the binding capacity and concentration dependence, which is same as that revealed in some experimental investigations. This indicates the necessity to take into account the binding capacity and chloride concentration dependence in the durability analysis and service life prediction of concrete structures.

염색법에 의한 쌀 단백질의 함량 및 질의 평가에 관한 연구 (Evaluation of Protein Content and Qaulity in Brown Rice by a Dye-binding Method)

  • 김성곤;이춘영;박훈
    • 한국식품과학회지
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    • 제3권2호
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    • pp.101-104
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    • 1971
  • Frankel-Conrat 와 Cooper에 의해 개발된 쉽고 빠르며 능률적으로 단백질 함량 및 질을 결정할 수 있는 염색법을 고단백 품종의 선발에 이용할 수 있는지의 여부를 알기 위하여 최근 고단백 수도품종으로 육종되고 있는 IR-667 계통에 적용하여 단백질, BAA, 및 라이신 함량과의 관계를 비교 검토하여 다음과 같은 결론을 얻었다. DBA-protein의 관계는 고도의 상관관계를 보였으며 단백질 함량을 생체중으로 나타냈을 때 $-0.975^{**}$, 건물중으로 나타냈을 때 $-0.955^{**}$의 상관계수를 보였다. DBA-lysine 및 DBA-BAA의 상관관계는 모두 고도의 상관관계를 보였으며 비슷한 상관계수를 보였다. DBA-lysine의 관계는 DBA-protein 의 관계보다는 다소 낮으나 protein-lysine과의 관계보다는 높은 상관관계를 나타내었다.

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잔류 Sulfamethazine 검출용 ELISA 개발에 관한 실험적 연구 (Experimental Study on Development of ELISA Method for the Detection of Sulfamethazine Residues)

  • 임윤규;김성희
    • 한국식품위생안전성학회지
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    • 제10권4호
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    • pp.213-217
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    • 1995
  • A screening method has been developed for detecting sulfamethazine(SMZ) contamination of meat or feeds by using horseradish peroxidase (HRP) labeled protein A (Prot AHRP)and an indirect competitve enzyme-linked immunosorbent assay(ELISA). The assay is based on competitve binding of guinea pig anti-SMZ with SMZ in smaple and SMZ-gelatin conjugate(SMZ.GEL). Percent binding (B.Bo$\times$100) was calculated from the absorbance in the absence (B0) and presence (B) of SMZ. By the sandard curve prepared by plotting log(SMZ) vs percent binding of each known reference solution, the detection limit was 1.0ppb or less. Cross reacton with sulfadimethoxine, sulfaguaniding, sulfamerazine, sulfamthoxpyridazine, sulfanilamide, sulfisomidine and sufisoxazole were not observed. But sulfamerazine crossreacted in the test. The EC-50 value (concentration causing 50% inhibition of color development compared with blank) of sulfamerazine was 2.0 ppm. Further quality control will make the ELISA system ideal for the detection of SMZ in meat or feeds.

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二中心 Huckel 法의 應用. 포화탄화수소의 C-C 切斷 및 水素의 反應性 (Application of Two Centre Huckel Method for C-C Bond Fission and Hydrogen Abstraction of Saturated Hydrocarbons)

  • 박병각;이무상
    • 대한화학회지
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    • 제16권2호
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    • pp.59-63
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    • 1972
  • In connection with two electron binding energy of each bond of saturated hydrocarbons, C-C bond fission and hydrogen abstraction from C-H bond are discussed by means of two center Huckel method. A beautiful correlation could be noticed between the observed bond dissociation energy and the calculated bond energy except for n-butane. Bond dissociation energies between C-C bond were also related to C-C bond fission. We could also find a very close relation between the relative easiness of hydrogen abstraction and the calculated binding energy of C-H bond. In other words, C-H bonds of tertiary hydrogen have been noticed as most weakely bonded and hence the tertiary hydrogen would most easily from the paraffins. In addition, the C-H binding energy is discussed applying ionic character of C-H bond which is derived from its dipole moment (0.4D)

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Enzyme-Linked, Biotin-Streptavidin Bacterial-Adhesion Assay for Helicobacter pylori Lectin-Like Interactions with Cultured Cells

  • Murillo, Guzman;Antonia, Maria;Ascencio, Felipe
    • Journal of Microbiology and Biotechnology
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    • 제11권1호
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    • pp.35-39
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    • 2001
  • A simple method for studying the lectin-like interactions between Helicobacter pylori and cultured human epithelial cell lines was developed using an enzyme-linked, biotin-streptavidin bacterial-adhesion assay. The present study suggests that this method is suitable for evaluating the participation of lectin interactions in the adhesion of H. pylori to cultured HeLa S3 and Kato III cells, both fixed and glycosidase-treated cells, as well as assessing glycoconjugated binding inhibition studies. The time-course and dose-dependent kinetics of the biotin-labeled H. pylori adhesion th the formaldehyde-fixed Hela S3 and Kato III cell lines exhibited saturation. In addition, the binding of the biotin-labeled H. pylori to the formaldehyde-fixed cultured cells was partially blocked by pre-incubation with glycoconjugates and polyclonal antibodies against a heparan sulfate binding protein from H. pylori.

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The Electronic Structure of Carbon Nanotubes with Finite Length : Tight Binding Theory

  • Moon, Won-Ha;Kim, Won-Woo;Hwang, Ho-Jung
    • Transactions on Electrical and Electronic Materials
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    • 제3권1호
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    • pp.23-29
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    • 2002
  • The electronic properties of Carbon Nanotube(CNT) are currently the focus of considerable interest. In this paper, the electronic properties of finite length effect in CNT for the carbon nano-scale device is presented. To Calculate the electronic properties of CNT, Empirical potential method (the extended Brenner potential for C-Si-H) for carbon and Tight Binding molecular dynamic (TBMD) simulation are used. As a result of study, we have known that the value of the band gap decreases with increasing the length of the tube. The energy band gap of (6,6) armchair CNT have the ranges between 0.3 eV and 2.5 eV. Also, our results are in agreements with the result of the other computational techniques.

Electronic Structure of Oxygen in the Defective Nickel Monoxide

  • 이광순;구현주;함경희;안운선
    • Bulletin of the Korean Chemical Society
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    • 제16권2호
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    • pp.164-168
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    • 1995
  • The band structure of nickel monoxide having a cation defect rock salt structure is calculated by means of the tight-binding extended Huckel method. The calculation is also made for the net charge, the DOS, the COOP, the electron density of the constituent atoms, and the O 1s binding energy shift when one of the adjacent nickel atoms is defected. It is found that the band gap near the Γ direction on the Brillouin zone is about 0.2 eV, and that all of the properties calculated including the electronic structure of the oxygen atom are more effectively affected by the surface defect than the inside one. The core O 1s binding energy shift is calculated by the use of valence potential method and the results are very satisfactory in comparison with the XPS experimental findings.

Molecular Dynamics Free Energy Simulation Study to Rationalize the Relative Activities of PPAR δ Agonists

  • Lee, Woo-Jin;Park, Hwang-Seo;Lee, Sangyoub
    • Bulletin of the Korean Chemical Society
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    • 제29권2호
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    • pp.363-371
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    • 2008
  • As a computational method for the discovery of the effective agonists for PPARd, we address the usefulness of molecular dynamics free energy (MDFE) simulation with explicit solvent in terms of the accuracy and the computing cost. For this purpose, we establish an efficient computational protocol of thermodynamic integration (TI) that is superior to free energy perturbation (FEP) method in parallel computing environment. Using this protocol, the relative binding affinities of GW501516 and its derivatives for PPARd are calculated. The accuracy of our protocol was evaluated in two steps. First, we devise a thermodynamic cycle to calculate the absolute and relative hydration free energies of test molecules. This allows a self-consistent check for the accuracy of the calculation protocol. Second, the calculated relative binding affinities of the selected ligands are compared with experimental IC50 values. The average deviation of the calculated binding free energies from the experimental results amounts at the most to 1 kcal/mol. The computational efficiency of current protocol is also assessed by comparing its execution times with those of the sequential version of the TI protocol. The results show that the calculation can be accelerated by 4 times when compared to the sequential run. Based on the calculations with the parallel computational protocol, a new potential agonist of GW501516 derivative is proposed.