• 제목/요약/키워드: Conjugated molecule

검색결과 52건 처리시간 0.025초

Single-Step Purification of Proteins of Interest from Proteolytically Cleaved Recombinant Maltose-binding Protein (MBP) Fusion Proteins by Selective Immunoprecipitation of MBP

  • Park, Jung-Hyun;Na, Shin-Young;Lee, Dong-Gun;Han, Byoung-Don;Kim, Kil-Lyong
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제3권2호
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    • pp.82-86
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    • 1998
  • The maltose binding protein (MBP) fusion protein system is a versatile tool to express and isolate recombinant proteins in E. coli. In this system, MBP fusion proteins are efficiently isolated from whole cell lysate using amylose conjugated agarose beads and then eluted by competition with free maltose. Since MBP is a rather large molecule (∼42 kDa), for further experiments, the MBP part is usually proteolytically cleaved from the fusion protein and subsequently removed by ion-exchange chromatography or rebinding to amylose columns after washing out excess and MBP-bound maltose. In the present study, we have developed an improved method for the removal of cleaved MBP, which is advantageous over conventional methods. In this method, factor Xa cleaved MBP fusion proteins were incubated with Sepharose beads conjugated with MBP specific monoclonal antibodies and then precipitated buy centrifugation, resulting in highly purified proteins in the supernatant.

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Cell-Specific Targeting of Texas Red with Anti-Ep-CAM Antibody

  • Lee, Soo-Chul;Tae, Gun-Sik
    • Journal of Photoscience
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    • 제12권3호
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    • pp.123-127
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    • 2005
  • The polyclonal antibody was generated against the peptide fragment of 62 amino acid residues (D 181-T242) near the COOH-terminal region of the extracellular domain of epithelial-cell adhesion molecule (Ep-CAM) and shown to be able to recognize Ep-CAM in competitive ELISA. Then, sulforhodamine 101 acid chloride (so called Texas red), a fluorescence dye, was conjugated to the affinity-purified anti-Ep-CAM antibody utilizing the reaction between the aliphatic amines of antibody and the sulfonyl chloride of Texas red. The molar ratio of Texas red to antibody was estimated to be approximately 1.86 by measuring optical densities at 280 nm and 596 nm, implying that the two molecules of Texas red at most were conjugated to antibody. The anti-Ep-CAM antibody-Texas red conjugate was then used for immunohistochemistry of CT-26 murine colon carcinoma cells. Based upon the fluorescence microscope images, anti-Ep-CAM antibody is able to deliver Texas red specifically to the surface of CT-26 cells on which Ep-CAM was actively expressed. This result indicates that anti-Ep-CAM antibody could be useful for the tissue-specific delivery of photosensitizers via antigen-antibody interaction.

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The Characteristics of Molecular Conjugated Optical Sensor Based on Silicon Nanowire FET

  • 이동진;김태근;황동훈;황종승;황성우
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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    • pp.486-486
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    • 2013
  • Silicon nanowire devices fabricated by bottom-up methods are attracted due to their electrical, mechanical, and optical properties. Especially, to functionalize the surface of silicon nanowires by molecules has received interests. The changes in the characteristics of the molecules is delivered directly to the surface of the silicon nanowires so that the silicon nanowire can be utilized as an efficient read-out device by using the electronic state change of molecules. The surface treatment of the silicon nanowire with light-sensitive molecules can change its optical characteristics greatly. In this paper, we present the optical response of a SiNW field-effect-transistor (FET) conjugated with porphyrin molecules. We fabricated a SiNW FET and performed porphyrin conjugation on its surface. The characteristic and the optical response of the device shows a large difference after conjugation while there is not much change of the surface in the SEM observation. It attributed to the existence of few layer porphyrin molecules on the SiNW surface and efficient variation of the surface potential of the SiNW due to light irradiation.

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In vitro and in vivo application of anti-cotinine antibody and cotinine-conjugated compounds

  • Kim, Hyori;Yoon, Soomin;Chung, Junho
    • BMB Reports
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    • 제47권3호
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    • pp.130-134
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    • 2014
  • The combination of a high-affinity antibody to a hapten, and hapten-conjugated compounds, can provide an alternative to the direct chemical cross-linking of the antibody and compounds. An optimal hapten for in vitro use is one that is absent in biological systems. For in vivo applications, additional characteristics such as pharmacological safety and physiological inertness would be beneficial. Additionally, methods for cross-linking the hapten to various chemical compounds should be available. Cotinine, a major metabolite of nicotine, is considered advantageous in these aspects. A high-affinity anti-cotinine recombinant antibody has recently become available, and can be converted into various formats, including a bispecific antibody. The bispecific anti-cotinine antibody was successfully applied to immunoblot, enzyme immunoassay, immunoaffinity purification, and pre-targeted in vivo radioimmunoimaging. The anti-cotinine IgG molecule could be complexed with aptamers to form a novel affinity unit, and extended the in vivo half-life of aptamers, opening up the possibility of applying the same strategy to therapeutic peptides and chemical compounds.

Development and Evaluation of Non-Hydrous Skin Analogue Liquid Crystal using Thermo-Sensitivity Smart Sensor

  • Yoo, Kwang-Ho;Hong, Jae-Hwa;Eun, So-Hee;Jeong, Tae-Hwa;Jeong, Kwan-Young
    • 한국응용과학기술학회지
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    • 제31권3호
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    • pp.367-374
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    • 2014
  • In this study, skin permeation enhancement was confirmed by designing it to have a structure and composition similarity to the intercellular lipids that improve miscibility with skin by cross-linked lipids poloxamer. The cross-linked lipids poloxamer was synthesized and analyzed by 1H NMR that structure dose had conjugated pluronic with ceramide3. Active component is released by modification of liquid crystal structure because PPO part, large-scale molecule block of pluronic, has hydrophobic nature at skin temperature of $35^{\circ}C$. Conjugated pluronic with ceramide3 was synthesized using Pluronic F127 and p-NPC (4-nitrophenyl chloroformate) at room temperature yielded 89%. Pluronic(Ceramide 3-conjugated Pluronic) was synthesized by reaction of p-NP-Pluronic with Ceramide3 and DMAP. The yield was 51%. This cross-linked lipids poloxamer was blended and dissolved at isotropic state with skin surface lipids, phospholipid, ceramide, cholesterol and anhydrous additive solvent. Next step was preceded by ${\alpha}$-Transition at low temperature for making the structure of Meso-Phase Lamella, and non-hydrous skin analogue liquid crystal using thermo-sensitivity smart sensor, lamellar liquid crystal structure through aging time. For confirmation of conjugation thermo-sensitivity smart sensor and non-hydrous skin analogue liquid crystal, structural observation and stability test were performed using XRD(Xray Diffraction), DSC(Differential Scanning Calorimetry), PM (Polarized Microscope) And C-SEM (Cryo-Scanning Electron Microscope). Thermo-sensitivity observation by Franz cell revealed that synthesized smart sensor shown skin permeation effect over 75% than normal liquid crystal. Furthermore, normal non-hydrous skin analogue liquid crystal that not applied smart sensor shown similar results below $35^{\circ}C$ of skin temperature, but its effects has increased more than 30% above $35^{\circ}C$.

Spectroscopic Properties of Quercetin in AOT Reverse Micelles

  • Park, Hyoung-Ryun;Im, Seo-Eun;Seo, Jung-Ja;Bark, Ki-Min
    • Bulletin of the Korean Chemical Society
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    • 제35권3호
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    • pp.828-832
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    • 2014
  • The spectroscopic properties of quercetin (QCT) were studied in the AOT reverse micelle by fluorescence spectroscopy. Because the molecular structure of QCT is completely planar, excited state intramolecular proton transfer (ESIPT) occurs between the -OH at C(5) and carbonyl oxygen via intramolecular hydrogen bonding. This ESIPT happens at the $S_1$ state but not at the $S_2$ state. Because QCT is a good donor-acceptor-conjugated molecule at the excited state, this molecule can emit strong fluorescence but shows no $S_1{\rightarrow}S_o$ emission due to this ESIPT. Since the $S_2{\rightarrow}S_1$ internal conversion was very slow due to the small Franck-Condon factors, $S_2{\rightarrow}S_o$ fluorescence emission was observed. All of the experimental results indicated that the QCT resided at the bound water interface and that the position of solute did not change significantly in the micelle at various water concentrations.

Dynamics of RNA Bacteriophage MS2 Observed with a Long-Lifetime Metal-Ligand Complex

  • Kang, Jung Sook;Yoon, Ji Hye
    • Journal of Photoscience
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    • 제11권1호
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    • pp.35-40
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    • 2004
  • [Ru(2,2'-bipyridine)$_2$(4,4'-dicarboxy-2,2'-bipyridine)]$^{2+}$(RuBDc) is a very photostable probe that possesses favorable photophysical properties including long lifetime, high quantum yield, large Stokes' shift, and highly polarized emission. To evaluate the usefulness of this luminophore (RuBDc) for studying macromolecular dynamics, its intensity and anisotropy decays when conjugated to RNA bacteriophage MS2 were examined using frequency-domain fluorometry with a high-intensity, blue light-emitting diode (LED) as the modulated light source. The intensity decays were best fit by a sum of two exponentials, and the mean intensity decay time was 442.2 ns. The anisotropy decay data showed a single rotational correlation time (2334.9 ns), which is typical for a spherical molecule. The use of RuBDc enabled us to measure the rotational correlation time up to several microseconds. These results indicate that RuBDc can be useful for studying rotational diffusion of biological macromolecules.s.

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Electronic structure of potassium-doped copper phthalocyanine studied by photoemission spectroscopy and density functional calculations

  • 임영지;김종훈;지동현;조상완
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2016년도 제50회 동계 정기학술대회 초록집
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    • pp.142.2-142.2
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    • 2016
  • The metal intercalation to an organic semiconductor is of importance since the charge transfer between a metal and an organic semiconductor can induce the highly enhanced conductivity for achieving efficient organic electronic devices. In this regard, the changes of the electronic structure of copper phthalocyanine (CuPc) caused by the intercalation of potassium are studied by ultraviolet photoemission spectroscopy (UPS) and density functional theory (DFT) calculations. Potassium intercalation leads to the appearance of an intercalation-induced peak between the highest molecular occupied orbital (HOMO) and the lowest molecular unoccupied orbital (LUMO) in the valence-band spectra obtained using UPS. The DFT calculations show that the new gap state is attributed to filling the LUMO+1, unlike a common belief of filling the LUMO. However, the LUMO+1 is not conductive because the ${\pi}$-conjugated macrocyclic isoindole rings on the molecule do not make a contribution to the LUMO+1. This is the origin of a metal-insulator transition through heavily potassium doped CuPc.

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Specific Targeting of Fluorescein Isothiocyanate with Ep-CAM Antibody(Specific targeting of FITC with Ep-CAM Antibody)

  • Lee, Young-Tae;Tae, Gun-Sik
    • Journal of Photoscience
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    • 제10권3호
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    • pp.237-240
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    • 2003
  • The tetradecameric peptide (K47-K60) near the NH$_2$-terminal region of epithelial-cell adhesion molecule (Ep-CAM) was chosen as antigenic site and a polyclonal antibody was generated, which could recognize Ep-CAM from the mouse colon tissue or the colon cancer cell, CT-26, in Western blot analysis. Then, the fluorescein isothiocyanate (FITC), a fluorescence dye, was conjugated with the affinity purified Ep-CAM antibody using thiocyanate and the amino groups of FITC and antibody, respectively. The molar ratio of FITC to antibody was estimated approximately 1.86 to 1.00 by measuring the optical densities at 492 nm and 280 nm. Ep-CAM antibody-FITC conjugate was then used for immunohistochemistry of the CT-26 cells. Judging from the shapes formed by fluorescence, the Ep-CAM antibody could delivered FITC to the surface of cells in which Ep-CAM was expressed. This result implies that Ep-CAM antibody could be also used for the tissue-specific delivery of the photosensitizer to the target protein via antigen-antibody interaction.

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Synthesis of Host Polymers and Guests for Electrophosphorescence

  • Holmes Andrew B.;Chan, Khai-Leok;Cho, Sung-Yong;Evans Nicholas R.;Grimsdale Andrew C.;Mak Chris S.K.;Sandee Albertus J.;Watkins Scott E.;Williams Charlotte K.
    • 한국고분자학회:학술대회논문집
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    • 한국고분자학회 2006년도 IUPAC International Symposium on Advanced Polymers for Emerging Technologies
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    • pp.21-22
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    • 2006
  • Significant progress has been realized in the design and synthesis of light emitting polymers with emission over the whole range of the visible spectrum. However up to seventy-five percent of charge recombination events can lead to triplet states that decay non-radiatively. Following the pioneering work in the field of small molecule organic light emitting devices it has been found that solution processible iridium polymer complexes can be used to harness the wasted triplet energy. In this paper new results concerning electrophosphorescence of solution processible tethered iridium polymer derivatives will be presented. Furthermore our approaches to the design of new high triplet energy conjugated polymer hosts will be reported.

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