• 제목/요약/키워드: affinity matrix

검색결과 95건 처리시간 0.024초

다양한 정제방법에 의한 전기영동용 한천유래 아가로즈의 제조 및 DNA분리 특성 (Preparation of Agarose from Gelidium amansii for Gel Electrophoresis using Various Purification Methods and Its Resolution Characteristics for DNA)

  • 도정룡;오세욱
    • 한국식품과학회지
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    • 제31권1호
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    • pp.110-114
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    • 1999
  • 제주도산 우뭇가사리로부터 agar를 제조하여 분석용 등급의 agarose 제조를 위하여 cetylpyridium chloride (CPC), polyethylene glycol (PEG), chitosan을 처리하여 agaropectin을 제거하여 agarose를 분리, 정제하였다. 3가지 방법중 chitosan을 처리하여 정제한 agarose는 황산기 함량이 가장 낮았으며, electroendosomosis도 가장 낮은 수치를 나타내어 분석용 agarose 제조를 위한 가장 효율적인 방법으로 나타났으며 실제 DNA 전기영동 실험에서도 가장 우수한 resolution을 나타내었으며 교차면역 전기영동에서도 시판되고 있는 제품에 비하여 품질이 양호함을 알 수 있었다.

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충전재의 종류에 따른 유리/노볼락 복합재료의 기계적 및 열적 성질 연구 (Effect of Fillers on the Mechanical and Thermal Properties of Glass/Novolac Composites)

  • 이수;이인규;박상희
    • 한국응용과학기술학회지
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    • 제25권1호
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    • pp.15-22
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    • 2008
  • The effects of fillers on the mechanical and thermal properties of glass/novolac composites have been studied. The matrix polymer and reinforcement were novolac type phenolic resin and milled glass fiber, respectively. Three different fillers, such as calcium carbonate, aluminum oxide, and wood powder were used for glass fiber reinforced plastic(GFRP) manufacture. Gravity, moisture content, tensile and flexural strength were measured to analyze the mechanical properties of GFRP and the final composites was burned in the electronic furnace at $1000^{\circ}C$ to confirm thermal properties GFRP containing aluminium oxide shows the highest thermal stability with 32% of weight loss at $1000^{\circ}C$ for one hour. GFRP containing calcium carbonate shows the maximum flexural strength (146 MPa), but that containing wood powder dose the highest tensile strength (65 MPa). Conclusively, we found that the characteristics of final composites strongly depend on several factors, such as types of materials, contents and chemical affinity of fillers. Therefore, it is very important to set up the combination of fillers for GFRP manufacturing to improve both mechanical and thermal properties at the same time.

FAP Inhibitors as Novel Small Molecules for Cancer Imaging using Radionuclide

  • Anvar Mirzaei;Jung-Joon Min;Dong-Yeon Kim
    • 대한방사성의약품학회지
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    • 제9권1호
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    • pp.49-55
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    • 2023
  • Tumors are encircled by various non-cancerous cell types in the extracellular matrix, including fibroblasts, endothelial cells, immune cells, and cytokines. Fibroblasts are the most critical cells in the tumor stroma and play an important role in tumor development, which has been highlighted in some epithelial cancers. Many studies have shown a tight connection between cancerous cells and fibroblasts in the last decade. Regulatory factors secreted into the tumor environment by special fibroblast cells, cancer-associated fibroblasts (CAFs), play an important role in tumor and vessel development, metastasis, and therapy resistance. This review addresses the development of FAP inhibitors, emphasizing the first, second, and latest generations. First-generation inhibitors exhibit low selectivity and chemical stability, encouraging researchers to develop new scaffolds based on preclinical and clinical data. Second-generation enzymes such as UAMC-1110 demonstrated enhanced FAP binding and better selectivity. Targeted treatment and diagnostic imaging have become possible by further developing radionuclide-labeled fibroblast activation protein inhibitors (FAPIs). Although all three FAPIs (01, 02, and 04) showed excellent preclinical and clinical findings. The final optimization of these FAPI scaffolds resulted in FAPI-46 with the highest tumor-to-background ratio and better binding affinity.

CaCO3을 이용한 혼합매질분리막의 이산화탄소 투과도 향상 (Enhancement of CO2 permeance by incorporating CaCO3 in Mixed Matrix Membranes)

  • 박철훈;정정표;이재훈;김종학
    • 멤브레인
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    • 제28권1호
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    • pp.55-61
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    • 2018
  • 급진적인 산업발전에 따른 무분별한 이산화탄소 배출로 인하여 지구온난화, 이상 기후 등의 문제가 불거지고 있으며 이는 한 나라에만 국한되지 않은 국제적인 문제가 되었다. 위와 같은 문제를 해결하고자 본 연구에서는 이산화탄소에 대해 높은 친화도를 가지고 있는 PEGBEM-g-POEM 가지형 공중합체를 합성하였으며 $CaCO_3$염을 첨가하여 염의 중량에 따른 혼합매질분리막의 이산화탄소/질소 투과 성능을 관찰하였다. $CaCO_3$염이 고분자 중량 대비 50% 첨가되었을 때 최적의 이산화탄소/질소 분리 투과성능을 보였으며, 선택도는 염을 추가하기 전 44.7에서 45.42로 크게 변하지 않은 반면 이산화탄소 투과도는 염을 첨가하기 전 22.5 GPU에서 28.16 GPU로 약 25% 향상되었다. 이는 첨가한 염과 이산화탄소간의 상호작용으로 인하여 향상된 고분자 복합막의 이산화탄소에 대한 용해도에 기인하는 것으로 생각된다. 또한 염을 과량으로 첨가한 경우에는 고분자가 복합막의 표면을 충분히 메우지 못하여 계면결함이 생성되었으며 이로 인해 이산화탄소 기체 분리성능이 감소하였다. 기체분리성능 향상을 위해서는 적정량의 염 첨가가 필수적인 것으로 분석되었다.

Generation of a monoclonal anti-human $\beta$2-adrenergic receptor antibody using GST-$\beta$-adrenergic receptor C-terminal fusion proteins expressed in E.Coli.

  • Kang, Suk-Jo;Shin, Chan-Young;Park, Kyu-Hwan;Ko, Kwang-Ho
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1997년도 춘계학술대회
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    • pp.95-95
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    • 1997
  • Among the various receptor molecules discovered so far the ${\beta}$2-adrenergic receptors have been regarded as excellent model systems for the so called 7 transmembrane helix receptor and have been the focus of extensive studies. For the analysis of receptor structure and function a monoclonal antibody plays a crucial role, thus providing useful tools for the study of receptor. However, because of the minute quantity of receptor molecules which could be obtained from natural sources, the generation of specific monoclonal antibody against receptor molecules from the purified receptors has been regarded as virtually impractical in consideration of cost and experimental times. The purpose of the present study was to generate and characterize a monoclonal antibody against human ${\beta}$2-adrenergic receptor. For the production of antibody, C-terminal regions of the human ${\beta}$2-adrenergic receptor was produced as a fusion protein with Glutathion S-transferase (GST) in E. Coli. The expression of the fusion protein was identified by SDS-PAGE and Western blot using monoclonal anti-GST antibody. The fusion protein was purified to an apparent homogeniety by affinity chromatography with Glutathion Sepharose CL-4B and used as an antigen for the immunization of BALB/C mice. The Production of monoclonal antibody was achieved by fusion of the immunized spleen cells and SP/2-0 myeloma cells. Positive hybridomas were screened by ELISA and were cloned by two consecutive rounds of limiting dilution. The monoclonal antibody produced in this study (mAb${\beta}$C02) was IgM type and purified by immunoaffinity chromatography using anti-mouse IgM agarose as an affinity matrix. MAb${\beta}$C02 showed strong and specific immunoreactivity against both the fusion protein and human ${\beta}$2-adrenergic receptor in ELISA and Western blot. The molecular weight of immunoreactive band was 64 kDa and exactly coincided with the previously reported molecular weight of ${\beta}$2-adrenergic recepters. The results of the present study suggest that mAb${\beta}$C02 may be used for the study of receptor function and regulation in normal or nonphysiological status.

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인체지방유래 간질세포의 부착 및 연골분화유도를 위한 PLGA 지지체의 플라즈마 처리 효과 (The Effect of the Plasma Treatment on PLGA Scaffold for Adhesion and Chondrogenic Differentiation of Human Adipose-derived Stromal Cells)

  • 동춘희;전영준;조현미;오득영;한동근;이종원;안상태
    • Archives of Plastic Surgery
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    • 제33권1호
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    • pp.46-52
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    • 2006
  • High-density micromass culture was needed to take three dimensions culture with ASCs(adipose derived stromal cells) and chondrogenesis. However, the synthetic polymer has hydrophobic character and low affinity to cells and other biomolecules. Therefore, the surface modification without changes of physical and chemical properties is necessary for more suitable condition to cells and biomolecules. This study was performed to investigate the effect of surface modification of poly (lactic-co-glycolic acid)(PLGA) scaffold by plasma treatment (P(+)) on the adhesion, proliferation and chondrogenesis of ASCs, and not plasma treatment (P(-)). ASCs were isolated from human subcutaneous adipose tissue obtained by lipectomy and liposuction. At 1 hour 30 minutes and 3days after cell seeding onto the P(-) group and the P(+) group, total DNA amount of attached and proliferated ASCs markedly increased in the P(+) group (p < 0.05). The changes of the actin under confocal microscope were done for evaluation of cellular affinity, at 1 hour 30 minutes, the shape of the cells was spherical form in all group. At 3rd day, the shape of the cells was fiber network form and finely arranged in P(+) group rather than in P(-) group. RT-PCR analysis of cartilage-specific type II collagen and link protein were expressed in 1, 2 weeks of induction. Amount of Glycoaminoglycan (GAG) markedly increased in P(+) group(p < 0.05). In a week, extracellular matrix was not observed in the Alcian blue and Safranin O staining. However in 2 weeks, it was observed that sulfated proteoglycan increased in P(+) group rather than in P(-) group. In conclusion, we recognized that plasma treatment of PLGA scaffold could increase the hydrophilic property of cells, and provide suitable environment for high-density micromass culture to chondrogenesis

Cloning, Over-expression, and Characterization of YjgA, a Novel ppGpp-binding Protein

  • Gnanasekaran, Gopalsamy;Pan, SangO;Jung, Wontae;Jeong, Kwangjoon;Jeong, Jae-Ho;Rhee, Joon Haeng;Choy, Hyon E.;Jung, Che-Hun
    • Bulletin of the Korean Chemical Society
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    • 제34권8호
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    • pp.2419-2424
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    • 2013
  • Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

초음파 수상 그래프팅을 이용하여 개질된 MWCNT가 첨가된 PVA 나노복합체의 전기적, 기계적 물성 (Mechanical and Electrical Properties of PVA Nanocomposite Containing Sonochemically Modified MWCNT in Water)

  • 김영선;김민재;최진규;심상은
    • 폴리머
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    • 제39권1호
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    • pp.136-143
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    • 2015
  • 초음파를 이용하여 poly(vinyl alcohol)(PVA)를 multiwalled carbon nanotube(MWCNT) 표면에 수상 그래프팅하였고, 개질된 MWCNT를 이용하여 PVA와 나노복합체를 제조하였다. PVA로 개질된 MWCNT는 PVA 매트리스에 높은 친화성을 띠고, 우수한 분산성을 가지며, 그 복합체는 균일한 물성을 가지고 있었다. 이로 인하여 0.1 wt% 함량의 MWCNT 첨가시에 전기전도도의 percolation threshold이 관찰되었다. 개질한 MWCNT를 3.0 wt%로 사용한 복합체는 순수 PVA 대비 인장강도는 약 50%, 파단 연신율은 약 430%, 모듈러스는 약 100% 증가하였다. 또한 개질된 MWCNT는 PVA 매트리스에 핵제로 작용하여 5.0 wt% 첨가 시, 결정화온도를 $8.5^{\circ}C$ 증가시키고 결정화도는 11.5% 증가하였다.

Effect of Quercetin in the UV-Irradiated Human Keratinocyte HaCaT Cells and A Model of Its Binding To p38 MAPK

  • Jnawali, Hum Nath;Lee, Eunjung;Shin, Areum;Park, Young Guen;Kim, Yangmee
    • Bulletin of the Korean Chemical Society
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    • 제35권9호
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    • pp.2787-2790
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    • 2014
  • Quercetin is a major dietary flavonoid found in onions, apples, tea, and red wine, and potentially has beneficial effects on disease prevention. We carried out this study to investigate the effect of quercetin on UVB-induced matrix metalloproteinase-1 (MMP-1) expression in human keratinocyte HaCaT cells and to further understand the mechanisms of its action. The anti-inflammatory activity of quercetin was investigated and quercetin significantly suppressed the NO production in LPS-stimulated RAW264.7 mouse macrophages. Post treatment of quercetin decreased UV irradiation-induced phosphorylation of JNK, p38 MAPK, and ERK by 91%, 21%, and 17%, respectively. MMP-1 is mainly responsible for the degradation of dermal collagen during the aging process of human skin and quercetin suppressed the UVB-induced MMP-1 by 94%. Binding studies revealed that quercetin binds to p38 with high binding affinity ($1.85{\times}10^6M^{-1}$). The binding model showed that the 4'-hydroxy groups of the B-ring of quercetin participated in hydrogen bonding interactions with the side chains of Lys53, Glu71, and Asp168 and the 5-hydroxy group of the A-ring formed a hydrogen bond with the backbone amide of Met109. The major finding of this study shows that quercetin inhibits phosphorylation of JNK, p38 MAPK, and ERK pathway leading to the prevention of MMP-1 expression in human keratinocyte HaCaT cells. Therefore, our findings suggested the potentials of quercetin as a skin anti-photoaging agent.

생분해성 다공질 Calcium Metaphosphate 블록의 조직적합성에 관한 연구 (Biocompatibility and Bone Conductivity of Porous Calcium Metaphosphate Blocks)

  • 이용무;김석영;신승윤;구영;류인철;정종평
    • Journal of Periodontal and Implant Science
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    • 제28권4호
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    • pp.559-568
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    • 1998
  • direct bone apposition during bone remodelling. To address these problem, we developed a new ceramic, calcium metaphosphate(CMP), and report herein the biologic response to CMP in subcutaneous tissue, muscle and bone. Porous CMP blocks were prepared by condensation of anhydrous $Ca(H_2PO_4)_2$ to form non-crystalline $Ca(PO_3)_2$. Macroporous scaffolds were made using a polyurethane sponge method. CMP block possesses a macroporous structure with approximate pore size range of 0.3-1mm. CMP blocks were implanted in 8mm sized calvarial defect, subcutaneous tissue and muscle of 6 Newzealand White rabbits and histologic observation were performed at 4 and 6 weeks later. CMP blocks in subcutaneous tissue and muscle were well adapted without any adverse tissue reaction and resorbed slowly and spontaneously. Histologic observation of calvarial defect at 4 and 6 weeks revealed that CMP matrix were mingled with and directly apposed to new bone without any intervention of fibrous connective tissue. CMP blocks didn't show any adverse tissue reaction and resorbed spontaneously also in calvarial defect. This result revealed that CMP had a high affinity for bone and was very biocompatible. From this preliminary result, it was suggested that CMP was a promising ceramic as a bone substitute and tissue engineering scaffold for bone formation.

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