• 제목/요약/키워드: heavy chain

검색결과 335건 처리시간 0.031초

Isolation and Characterization of Single-Chain Fv Against Ductal Cells

  • Lee Myung-Hoon;Ryu Hye-Myung;Kim Sun-Zoo;Park Ji-Young;Uhm Ji-Hyun;Park Tae-In
    • 대한의생명과학회지
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    • 제10권3호
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    • pp.211-217
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    • 2004
  • For discrimination of ductal and ascinar cells, we isolated a single-chain variable domain fragment (scFv) antibody against ductal cells of salivary gland using phage display technique. From the spleen of a mouse immunized with ductal cell lysate, total RNA was prepared and used as a template for cDNA synthesis of antibody genes. The scFv genes were constructed with variable domain genes of heavy and light chain and were introduced into pCANTAB5E to construct phage scFv library. The phage particles specific for acinar cells were screened by subtraction using immunotubes coated with acinar and ductal cell lysate and enzyme-linked immunoabsorbance assay (ELISA). The characteristics of the scFv were determined by immunohistochemistry (IHC) and the result indicated that the isolated scFv has the specificity against ductal cells of salivary glands and tubules of kidney. And the scFv has an unique binding activity specific for Hashimoto's thyroiditis. The nucleotide sequence of isolated scFv gene was determined and revealed that V/sub H/ belongs to the mouse H-chain family subgroup IB and V/sub L/ to the mouse L-chain family subgroup III.

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설계 개선을 통한 선박의 계류 시스템 최적화 사례 소개 (Introduction of Optimized Design of Anchoring System through Design Modification of Pocket and Chain Compressor)

  • 이재훈
    • 대한조선학회 특별논문집
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    • 대한조선학회 2011년도 특별논문집
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    • pp.55-62
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    • 2011
  • Although the performance of the commercial vessel has been dramatically improved through innovations, there has been no big changes on the traditional anchoring method of commercial vessels, both on design and operation until now. In this paper, two cases of design modifications were introduced for optimized design of pocket type anchor handling, which resulted in improved performance of the vessel's anchoring. From the first time fully balanced type anchors were applied on vessels in Korean shipyard, main design problem on this application was that the anchor doesn't normally slide into the pocket when the anchor fluke is not in line with pocket, as the anchor freely rotates by the swivel on forerunner. In order to prevent the problem, swivel has been deleted on the forerunner to prevent anchor rotation until now, but this solution caused problems such as twist lock of anchor chain, restriction of windlass direction, etc. On this paper, one of the solution is introduced to overcome the design problem by tilting the hawse pipe to some extent, which makes anchor turned at the time anchor ring touches the pocket skirt and that it properly slides into the pocket. Secondly, one of the solution is introduced to overcome misalignment problem between anchor chain cable and roller of chain compressor, which has been frequently occurred, by modification of roller design.

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Generation of 1E8 Single Chain Fv-Fc Construct Against Human CD59

  • Hong, Jeong-Won;Cho, Woon-Dong;Hong, Kwon-Pyo;Kim, So-Seul;Son, Seung-Myoung;Yun, Seok-Joong;Lee, Ho-Chang;Yoon, Sang-Soon;Song, Hyung-Geun
    • IMMUNE NETWORK
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    • 제12권1호
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    • pp.33-39
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    • 2012
  • Background: Therapeutic approaches using monoclonal antibodies (mAbs) against complement regulatory proteins (CRPs:i.e.,CD46,CD55 and CD59) have been reported for adjuvant cancer therapy. In this study, we generated a recombinant 1E8 single-chain anti-CD59 antibody (scFv-Fc) and tested anti-cancer effect.by using complement dependent cytotoxicity (CDC). Methods: We isolated mRNA from 1E8 hybridoma cells and amplified the variable regions of the heavy chain (VH) and light chain (VL) genes using reversetranscriptase polymerase chain reaction (RT-PCR). Using a linker, the amplified sequences for the heavy and light chains were each connected to the sequence for a single polypeptide chain that was designed to be expressed. The VL and VH fragments were cloned into the pOptiVEC-TOPO vector that contained the human CH2-CH3 fragment. Then, 293T cells were transfected with the 1E8 single-chain Fv-Fc (scFv-Fc) constructs. CD59 expression was evaluated in the prostate cancer cell lines using flow cytometry. The enhancement of CDC effect by mouse 1E8 and 1E8 scFv-Fc were evaluated using a cytotoxicity assay. Results: The scFv-Fc constructs were expressed by the transfected 293T cells and secreted into the culture medium. The immunoreactivity of the secreted scFv-Fc construct was similar to that of the mouse 1E8 for CCRF-CEM cells. The molecular masses of 1E8 scFv-Fc were about 120 kDa and 55 kDa under reducing and non-reducing conditions, respectively. The DNA sequence of 1E8 scFv-Fc was obtained and presented. CD59 was highly expressed by the prostate cancer cell line. The recombinant 1E8 scFv-Fc mAb revealed significantly enhanced CDC effect similar with mouse 1E8 for prostate cancer cells. Conclusion: A 1E8 scFv-Fc construct for adjuvant cancer therapy was developed.

절대절점좌표를 이용한 회전체인의 시뮬레이션 (Simulation of a Rotating Chain with an Absolute Nodal Coordinate Formulation)

  • 유완석
    • 대한기계학회:학술대회논문집
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    • 대한기계학회 2004년도 추계학술대회
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    • pp.649-654
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    • 2004
  • A physically simple but mathematically cumbrous problem of rotating heavy chain with one fixed top point is studied. Nonlinear equation of its two-dimensional shapes of relative equilibrium is obtained and solved numerically. A linear case of small displacements is analyzed in terms of Bessel functions. The qualitative and quantitative behavior of the problem is discussed with the help of bifurcation diagram. Dynamics of the two-dimensional model near the equilibrium positions is studied with the help of simulation using the absolute nodal coordinate formulation (ANCF). The equilibriums are found instable, and the reason of instability is explained using a variational principle.

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Assessment of Selected Heavy Metal Concentrations in Agricultural Soils around Industrial Complexes in Southwestern Areas of Korea

  • Kim, Dong-Jin;Park, Jung-Hwon;Lee, Jin-Ho
    • 한국토양비료학회지
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    • 제49권5호
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    • pp.524-530
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    • 2016
  • Agricultural soils near or around industrial complexes can contain a certain amount of heavy metals that readily enter the food chain and negatively affect human health. Therefore, we conducted the study to investigate the distribution of selected heavy metals, including arsenic (As), cadmium (Cd), chromium (Cr), nickel (Ni), lead (Pb), mercury (Hg), and zinc (Zn), in farm-land soils around fifteen industrial complexes in the southwestern provinces, Korea. The concentrations of heavy metals in the soil samples were determined by the pseudo-total aqua regia (3 HCl : $1HNO_3$) digestion procedure. The heavy metal concentrations in most soils examined did not exceed the levels of Soil Contamination Warning Standard (SCWS) for agricultural lands (Region 1) presented in Soil Environment Conservation Law (SECL) established by Ministry of Environment (MOE), Korea. However, only one sampling site showed higher As amount ($27.1mg\;kg^{-1}$) than the SCWS level of As ($25mg\;kg^{-1}$). Pollution index (PI) for heavy metals did not exceed 1.0. The PI values were significantly positively correlated (p < 0.01) with the heavy metal concentrations. In particular, the values of correlation coefficient between the Cd and Pb concentrations and the PI values were higher than those estimated from other combinations, and thus the amounts of Cd and Pb in the agricultural soils highly affected the PI values for the heavy metals.

도인(桃仁)이 중대뇌동맥 폐색 유발 흰쥐의 근육형태 변화와 혈관신생에 미치는 영향 (Muscle Type Change and Vascularization Effect of Semen Persicae in MCAo rats)

  • 권혁준;소갑석;김호준;이명종
    • 한방재활의학과학회지
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    • 제19권1호
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    • pp.11-21
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    • 2009
  • Objectives : This study was performed to investigate the effects of Semen Persicae(SP) on muscle type change and vascularization effect by measuring expression of Myosin heavy chain (MHC) and Vascular endothelial growth factor (VEGF) protein in the Middle cerebral artery occlusion(MCAo) rats. Methods : The middle cerebral artery was occluded, SP extraction was administrated for 4 days. The effects of SP on muscle type change and vascularization were measured. Results : 1. VEGF protein expressions in the Semen Persicae oral administration group of MCAo group were increased compared to the control group. 2. There were no significant difference between the Semen Persicae oral administration of MCAo group and the control group in MHC isoform (Type I, Type IIa) expression change. Conclusions : The present study demonstrates the effect of Semen Persicae in the vascularizing after ischemia, but has no significant effect in musle type change and the improvement of musle atrophy.

DBcAMP와 retinoic acid를 이용한 마우스 배아줄기의 평활근세포 분화 (Differentiation of mouse embryonic stem cell into smooth muscle cells by DBcAMP and retinoic acid)

  • 박성수;강주원
    • 한국동물위생학회지
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    • 제31권4호
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    • pp.449-456
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    • 2008
  • The differentiation of mouse embryonic stem(ES) cell into smooth muscle cells(SMC) may play a major role in cardiovascular development and under pathophysiological conditions. Therefore, in the present study, we have examined the differentiation of ES cells and its related gene expression. SMC differentiation was indicated by cellular morphology and time-dependent induction of dibutyryl adenosine 3,5-cyclic monophosphate(DBcAMP)and retinoic acid(RA) on smooth muscle ${\alpha}$-actin($SM{\alpha}A$), smooth muscle myosin heavy chain(SMMHC) gene expression. The control was undifferentiated ES cells(protein expressions represent 50-60kDaOct-4). The results of this study show that morphology of embryoid body and confirmation of $SM{\alpha}A$ expression by immunocytochemistry. Moreover, SMMHC and desmin expression was significantly increased by time dependent manner(5, 7, 15 days), in contrast to $SM{\alpha}A$ expression was slightly decreased on 15days. In conclusion, DBcAMP and RA stimulate mouse ES cells differentiation into SMC and enhanced $SM{\alpha}A$, SMMHC and desmin expression.

A Method for Direct Application of Human Plasmin on a Dithiothreitol-containing Agarose Stacking Gel System

  • Choi, Nack-Shick;Chung, Dong-Min;Yoon, Kab-Seog;Maeng, Pil-Jae;Kim, Seung-Ho
    • BMB Reports
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    • 제38권6호
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    • pp.763-765
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    • 2005
  • A new simplified procedure for identifying human plasmin was developed using a DTT copolymerized agarose stacking gel (ASG) system. Agarose (1%) was used for the stacking gel because DTT inhibits the polymerization of acrylamide. Human plasmin showed the lowest activity at pH 9.0. There was a similar catalytically active pattern observed under acidic conditions (pH 3.0) to that observed under alkaline conditions (pH 10.0 or 11.0). Using the ASG system, the primary structure of the heavy chain could be established at pH 3.0. This protein was found to consist of three fragments, 45 kDa, 23 kDa, and 13 kDa. These results showed that the heavy chain has a similar structure to the autolysed plasmin (Wu et al., 1987b) but there is a different start amino acid sequence of the N-termini.

Production and characterization of monoclonal antibodies (MAb) against flounder serum immunoglobulin (Ig)

  • Jang, Han-Na;Cho, Young-Hye;Park, Sang-Hoon
    • 한국어업기술학회:학술대회논문집
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    • 한국어업기술학회 2000년도 춘계수산관련학회 공동학술대회발표요지집
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    • pp.446-446
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    • 2000
  • Specific polyclonal and/or monoclonal antibodies (MAbs) to immunoglobulins (Igs) and their subunits have proved to be valuable tools in immunological research and in immunological assays. In this study, we developed and characterized MAbs against flounder serum Igs. To obtain the pure flounder serum Igs, mouse IgG (mIgG) was immunized to flounder. Flounder Igs were purified by using mIgG-agarose affinity column chromatography. The structure of purified flounder Ig was observed, on denatured SDS-PAGE, to be composed of two heavy chains (77 and 72 kd) and two light chains (28 and 26 kd). MAbs were produced by fusion of myeloma cells (SP2/0) with Balb/c mouse spleen cells previously primed with the flounder Igs. Finally, three hybridoma clones, FIM 511, FIM 519 and FIM 562 were established to recognize both 2 heavy chains, 26 kd of light chain and 28 kd of light chain, respectively. On the other hand, the flounder immune sera collected on the weekly basis were tested on ELISA and immunoblot analysis whether boosting effect is present in flounder humoral immune system. As a result, the secondary immune response in flounder was ascertained on ELISA, but not on immunoblot analysis. Further, we observed an alteration of serum protein levels following immunization. Our MAbs and basic information on flounder humoral immune system obtained in this study will be helpful to control and monitor the efficiency of fish vaccines and therapeutic process of flounder diseases.

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