• Title/Summary/Keyword: affinity matrix

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Production of polyclonal anti-$\beta$-adrenergic receptor antibody and it′s effects on receptor ligand binding

  • Kim, Hee-Jin;Shin, Chan-Young;Noh, Min-Su;Ko, Kwang-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.86-86
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    • 1995
  • The analysis of membrane receptors for hormones and neurotransmitters has progressed considerably by pharmacological and biochemical means and more recently by the use of specific anti-receptor antibodies. A 14-mer peptide (from Phe102 to Leu115 of ${\beta}$2-adrenergic receptor) was synthesized and this peptide was coupled to carrier protein Keyhole Limpet Hemocyanin(KLH) by glutaraldehyde method. A 0.5mg of KLH-coupled peptide was emulsified with equal volume of complete Freund's adjuvant and injected via popliteal lymph node to each of the three Newzealnd White rabbits. Booster injections were repeated at 4 weeks interval for three times with incomplete Freund's adjuvants. One week after the final injection, serum was prepared from ear artery. Nonspecific immunoglobulins were removed by passing the serum through KLH-Sepharose 6B affinity matrix and further by incubation with bovine lung aceton powder. The titer of the antibody for synthetic peptide which was determined by enzyme linked immunosorbent assay(ELISA) was about l/l,000. The antibody produced in this study revealed 67kDa protein band in the western blot of partially purified guinea pig lung ${\beta}$-adrenergic receptor preparation. The antibody inhibited ${\beta}$-adrenergic antaginist [3H] Dihydroalprenolol binding to soluble ${\beta}$-adrenergic receptor by 25% while control sera did not show any inhibitory effects, The result of this study suggests that the peptide sequence selected in this study may play some important roles in adrenergic receptor-ligand interaction.

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Effect of Coupling Agent and Fiber Loading on Mechanical Behavior of Chopped Jute Fiber Reinforced Polypropylene Composites (황마 단섬유 강화 폴리프로필렌 복합재료의 기계적 거동에 미치는 결합제 및 섬유 Loading의 영향)

  • Rasel, S.M.;Nam, G.B.;Byeon, J.M.;Kim, B.S.;Song, J.I.
    • Elastomers and Composites
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    • v.46 no.3
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    • pp.204-210
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    • 2011
  • In this study, Jute fibers reinforced polypropylene (JFRP) composites were manufactured by injection molding technique. In order to improve the affinity and adhesion between fibers and thermoplastic matrices during manufacturing, Maleic anhydride (MA) as a coupling agent have been employed. Untreated and treated surfaces of jute fibers were characterized using SEM and Fourier transform infrared (FTIR). Physical properties like water absorption rate were studied. Tensile and flexural tests were carried out to evaluate the composite mechanical properties. Tensile test and bending test indicated that JFRP composites show higher strength and modulus than pure PP. In addition, strength and modulus were found to be influenced by the variation of MAPP content (1%, 2%, and 3%). Tensile fracture surfaces were examined using scanning electron microscope. It ensures better interfacial adhesion between fibers and matrix by increasing the percentage of MAPP.

EXPRESSIONS OF VASCULAR METASTASIS RELATED FACTORS IN MURINE ORTHOTOPIC TUMOR MODELS OF SALIVARY GLANDS (마우스에서 확립된 타액선 동위종양에서 혈관성 전이관련 인자의 발현)

  • Jang, Jae-Hyun;Kwon, Kwang-Jun;Park, Young-Wook
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.6
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    • pp.499-508
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    • 2007
  • Background and Purpose: Some subtypes of malignant salivary gland tumors such as adenoid cystic carcinoma (ACC) frequently result in distant metastasis of vascular origin, which are main causes of treatment failure. The reasons for the affinity for vascular metastatic potential are unclear. Therefore, molecular characteristics that influence the dissemination of metastatic tumor cells are important for the design of more effective treatment of salivary ACC. Tumor angiogenesis has been known to be essential for the distant metastasis of malignant cells. So, we determined expressions of vascular metastasis related factors in orthotopic (parotid) murine models of parotid ACC and compared with those in ectopic (subcutis) tumors of athymic mice. Experimental Design: Using specimens from murine parotid (orthotopic, experimental group) and subcutaneous (ectopic, control group) tumors, which have developed via transplantation of tumor cells, originated from human parotid ACC, we performed immunohistochemical assays with anti-vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF, FGF2), matrix metalloproteinase (MMP)-9, and interleukin (IL)-8 antibodies. We also performed immunohistochemical assays with VEGF receptor (VEGFR)-1, VEGFR-2, VEGFR-3, and phosphorylated VEGFR-2. Results: Transplantation of human ACC tumor cell $(5{\times}10^5)$ into the parotid and subcutis successfully resulted in orthotopic (parotid) and ectopic (subcutaneous) tumors in athymic mice. Immunohistochemical staining demonstrated higher expression of major angiogenic factors (VEGF, bFGF, MMP-9) in the orthotopic tumors than in ectopic tumors (P<0.05). But the expression level of angiogenic receptors were same in orthotopic and ectopic tumors of parotid ACC. Conclusion: VEGF, bFGF, and MMP-9 could be a good candidates for antiangiogenic therapy for the contol of vascular metastatic lesions of salivary ACC.

Surface Oxidation of High Strength Automotive Steels during Continuous Annealing, and the Influence of Trace Elements of P,B, and Sb

  • Sohn, Il-Ryoung;Park, Joong-Chul;Kim, Jong-Sang
    • Corrosion Science and Technology
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    • v.9 no.6
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    • pp.259-264
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    • 2010
  • In continuous hot dip galvanizing process, oxide formation on steel surface has an influence on Zn wetting. High strength automotive steel contains high amount of Si and Mn, where Si-Mn composite oxides such as $Mn_2SiO_4$ or $MnSiO_3$ covers the surface after annealing. Zn wetting depends on how the aluminothermia reaction can reduce the Mn-Si composite oxides and then form inhibition layer such as $Fe_2Al_5$ on the steel surface. The outward diffusion of metallic ions such as $Mn^{2+}$, $Si^{2+}$ in the steel matrix is very important factor for the formation of the surface oxides on the steel surface. The surface state and grain boundaries provide an important role for the diffusion and the surface oxide reactions. Some elements such as P, Sb, and B have a strong affinity for the interface precipitation, and it influence the diffusivity of metallic ions on grain boundaries. B oxide forms very rapildly on the steel surface during the annealing, and this promote complex oxides with $SiO_2$ or MnO. P has inter-reacted with other elements on the grain boundaries and influence the diffusion through on them. Small addition of Sb could suppress the decarburization from steel surface and retards the formation of internal and external selective oxides on the steel surface. Interface control by the trace elements such as Sb could be available to improve the Zn wettability during the hot dip galvanizing.

In Vitro Refolding of Inclusion Body Proteins Directly from E. coli Cell Homogenate in Expanded Bed Adsorption Chromatography (Expanded Bed Adsorption 크로마토그래피를 사용하여 재조합 E. coli 세포 파쇄액으로부터 내포체 단백질을 직접 재접힘하는 공정)

  • 조태훈;서창우;이은규
    • KSBB Journal
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    • v.16 no.2
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    • pp.146-152
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    • 2001
  • To avoid the intrinsic problem of aggregation associated with the traditional solution-phase refolding process, we propose a solid-phase refolding method integrated with expanded bed adsorption chromatography. The model protein used was a fusion protein of recombinant human growth hormone and a glutathione S transferase fragment. It was demonstrated that the EBA-mediated refolding technique could simultaneously remove cellular debris and directly renature the fusion protein inclusion bodies in the cell homogenate with much higher yields and less agregation. To demonstrate the applicability of the method, we successfully tested the three representative types of starting materials, i. e., rhGH monomer, washed inclusion bodies, and the E. coli homogenate. This direct and simplified refolding process could also reduce the number of renaturation steps required and allow refolding at a higher concentration, at approximately 2 mg fusion protein per ml of resin. To the best of our knowledge, it is the first approach that has combined the solid-phase refolding method with expanded bed chromatography.

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A New Process for the Requirements Based Aerospace System Design and Optimization (요구도 기반 항공우주 시스템 강건최적설계 기법 연구)

  • Park, Hyeong-Uk;Lee, Jae-Woo;Byun, Yung-Hwan;Chung, Joon;Behdinan, Karman
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.37 no.3
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    • pp.255-266
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    • 2009
  • In this study, a robust aerospace system design process for the aerospace system is developed by considering the uncertainties of user requirements, manufacturing errors, and operational environment variation. User requirements are analyzed and quantified by decision making models and system engineering methods to select alternative concepts which satisfies the various requirements. Robust design and optimization method is applied to derive the robust solution of the selected system. First, a variance of objective function is calculated, and a mean value and a variance of target value are determined by the deterministic design optimization results of the system. A robust optimum design formulation is then needed to derive the robust solution that minimizes the variance of the response and moves the mean values to the target value. It is applied to Very Light Jet (VLJ) aircraft to which much attention is paid recently in civil aerospace market.

Immobilization and properties of Streptomyces sp. S56 endoinulase (Streptomyces sp. S56 endoinulase의 고정화 및 성질)

  • Kim, Su-Il;Chang, Ho-Jin
    • Applied Biological Chemistry
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    • v.35 no.5
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    • pp.410-414
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    • 1992
  • Endoinulase from Streptomyces sp. S56 was immobilized by adsorption on DEAE-cellulose in 0.01 M citrate-sodium phosphate buffer, pH 6.0 and the properties of immobilized and free enzymes were investigated. The immobilized enzyme preparation, having 40 inulase activity units per dried matrix, revealed the maximal activity at $pH\;4.5{\sim}5.5$ and $55{\sim}60^{\circ}C$ and were most stable at pH 6 and 45^{\circ}C$. The immobilization caused a drop in optimum pH and affinity toward inulin, a slight increase in optimum temperature, an important increase in thermal stability and maximum reaction velocity. The immobilized endoinulase hydrolyzed the tuber extract of jerusalem artichoke and inulin, mainly into fructose and inulobise, degrading 63 and 78% of the total sugar respectively, within 48 hrs in batch reactor.

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Improving Smoothness of Hydrophilic Natural Polymer Coating Layer by Optimizing Composition of Coating Solution and Modifying Chemical Properties of Cobalt-Chrome Stent Surface (코팅 용액의 조성 최적화 및 코발트-크롬 금속스텐트의 화학적 표면개질을 통한 친수성 천연 고분자 코팅층의 표면 거칠기 개선)

  • Kim, Dae Hwan;Kum, Chang Hun
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.256-261
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    • 2018
  • Recently, the number of cardiovascular disease-related deaths worldwide has increased. Therefore, the importance of percutaneous cardiovascular intervention and drug-eluting stents (DES) has been highlighted. Despite the great clinical success of DES, the re-endothelialization at the site of stent implantation is retarded owing to the anti-proliferative effect from the coated drug, resulting in late thrombosis or very late restenosis. In order to solve this problem, studies have been actively carried out to excavate new drugs that promote rapid re-endothelialization. In this study, we introduced hydrophilic drug, tauroursodeoxycholate (TUDCA), that improves the proliferation of endothelial progenitor cells and promotes apoptosis of vascular smooth muscle cells. In addition, we utilized shellac, which is a natural resin from lac bug to coat TUDCA on the surface of the metal. When using conventional coating method including biodegradable polymers and organic solvents, phase separation between polymer and drug occurred in the coating layer that caused incomplete incorporation of drug into the polymer layer. However, when using shellac as a coating polymer, no phase separation was observed and drug was fully covered with the polymer matrix. In addition, by adjusting the composition of coating solution and modifying the hydrophilicity of the metal surface using oxygen plasma, the surface roughness decreased due to the increased affinity between coating solution and metal surface. This result provides a method of depositing a hydrophilic drug layer on the stent.

Preparation and Bioevaluation of 177Lu-labelled Anti-CD44 for Radioimmunotherapy of Colon Cancer

  • Lee, SoYoung;Hong, YoungDon;Jung, SungHee;Choi, SunJu
    • Journal of Radiation Industry
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    • v.9 no.4
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    • pp.187-192
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    • 2015
  • CD44 is a particular adhesion molecule and facilitates both cell-cell and cell-matrix interactions. In particular, splice variants of CD44 are particularly overexpressed in a large number of malignancies and carcinomas. In this study, the $^{177}Lu$-labelled CD44 targeting antibody was prepared and bioevaluated in vitro and in vivo. Anti-CD44 was immunoconjugated with the equivalent molar ratio of cysteine-based DTPA-NCS and radioimmunoconjugated with $^{177}Lu$ at room temperature within 15 minutes. The stability was tested in human serum. An in vitro study was carried out in HT-29 human colon cancer cell lines. For the biodistribution study $^{177}Lu$-labelled anti-CD44 was injected in xenograft mice. Anti-CD44 was immunoconjugated with cysteine-based DTPA-NCS and purified by a centricon filter system having a molecular cut-off of 50 kDa. Radioimmunoconjugation with $^{177}Lu$ was reacted for 15 min at room temperature. The radiolabeling yield was >99%, and it was stable in human serum without any fragmentation or degradation. The radioimmunoconjugate showed a high binding affinity on HT-29 colon cancer cell surfaces. In a biodistribution study, the tumor-to-blood ratio of the radioimmunoconjugate was 43 : 1 at 1 day post injection (p.i) in human colon cancer bearing mice. The anti-CD44 monoclonal antibody for the targeting of colon cancer was effectively radioimmunoconjugated with $^{177}Lu$. The in vitro high immunoactivity of this radioimmunoconjugate was determined by a cell binding assay. In addition, the antibody's tumor targeting ability was demonstrated with very high uptake in tumors. This radioimmunoconjugate is applicable to therapy in human colon cancer with highly expressed CD44.

Anti-inflammatory effects of Hataedock with Coptidis Rhizoma and Glycyrrhiza Uralensis on Allergic Rhinitis through Regulating IL-4 Activation (알레르기성 비염에서 황련-감초 하태독법의 IL-4활성 조절을 통한 항염증효과)

  • Jung, A Ram
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.33 no.2
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    • pp.116-122
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    • 2019
  • The aim of this study is to evaluate the anti-inflammatory effect of Hataedock treatment using Coptidis Rhizome and Glycyrrhiza Uralensis (CG) mixed extract in allergic rhinitis induced NC/Nga mice. We divided NC/Nga mice into 3 groups as follows; allergic rhinitis-induced group after CG Hataedock treatment (CGT, n=10), no treatment group (Ctrl), allergic rhinitis elicited group (ARE). To induce allergic rhinitis, NC/Nga mice of 3 weeks age were sensitized on 7, 8 and 9week by Ovalbumin (OVA) antigen in intranasal space. Hataedock using CG extract was administered on week 3 in allergic rhinitis-induced group (CGT) after Hataedock treatment. To identify distribution of Interlukin (IL)-4, Cluster of differentiation 40 (CD40), high-affinity IgE receptor ($Fc{\varepsilon}RI$), substance P, Matrix metallopeptidase 9 (MMP-9), Nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) p65, Inducible nitric oxide synthase (iNOS) and Cycloxygenase-2 (COX-2), we used histological examination. CGT significantly inhibited IL-4 and CD40 response compared with ARE. The reduction of Th2 cytokine expression decreased inflammatory mediators such as $Fc{\varepsilon}RI$, substance P, MMP-9, $NF-{\kappa}B$ p65, iNOS and COX-2. Such immunological improvement induced reduction of respiratory epithelial damage and mucin secretion in goblet cell. These results indicate that Hataedock treatment suppresses allergic rhinitis through modulating of Th2 responses and diminishing various inflammatory mediators in nasal mucosal tissue. It might have potential applications for prevention and treatment of allergic rhinitis.