• Title/Summary/Keyword: Tandem cell

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Development of Chromatographic Downstream Processing for The Purification of Monoclonal Antibody from Ascites Fluid: Part 1. Tandem Use of Hydroxylapatite Chromatography and Gel Permeation Chromatography (복수로부터의 단세포군 항체의 정제를 위한 크로마토그래프 분리 정제 시스템의 개발 - 1. 히드록실 아파타이트 크로마토그라프와 겔 여과 크로마토그라프로 이루어진 2단계 연속공정 사용 -)

  • Ahn, I.S.;Park, C.Y.
    • Microbiology and Biotechnology Letters
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    • v.17 no.1
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    • pp.19-23
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    • 1989
  • A sequential system composed of hydroxylapatite chromatography and gel permeation chromatography was developed to purify the IgM type monoclonal antibody against the colon cancer cell SC-1 from the ascitic fluid of mice injected with the murine hybridoma CH07E02. In the hydroxylapatite chromatographic step the band dilution could be reduced by controlling the gradient and flow rate of the eluent, the sodium phospate buffer, the optimum values for these variables being 5.82$\times$10$^{-3}$M/cm and 0.2$m\ell/\textrm{cm}^2$/min, respectively. A degree of purity better than 99.99% as judged from silverstaining of the SDS-PAGE bands, was obtained by adding the gel permeation chromatographic step in tandem.

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Identification of Cisplatin-Resistance Associated Genes through Proteomic Analysis of Human Ovarian Cancer Cells and a Cisplatin-resistant Subline

  • Zhou, Jing;Wei, Yue-Hua;Liao, Mei-Yan;Xiong, Yan;Li, Jie-Lan;Cai, Hong-Bing
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6435-6439
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    • 2012
  • Chemoresistance to cancer therapy is a major obstacle to the effective treatment of human cancers with cisplatin (DDP), but the mechanisms of cisplatin-resistance are not clear. In this study, we established a cisplatin-resistant human ovarian cancer cell line (COC1/DDP) and identified differentially expressed proteins related to cisplatin resistance. The proteomic expression profiles in COC1 before and after DDP treatment were examined using 2-dimensional electrophoresis technology. Differentially expressed proteins were identified using matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and high performance liquid chromatography-electrospray tandem MS (NanoUPLC-ESI-MS/MS). 5 protein spots, for cytokeratin 9, keratin 1, deoxyuridine triphosphatase (dUTPase), aarF domain containing kinase 4 (ADCK 4) and cofilin1, were identified to be significantly changed in COC1/DDP compared with its parental cells. The expression of these five proteins was further validated by quantitative PCR and Western blotting, confirming the results of proteomic analysis. Further research on these proteins may help to identify novel resistant biomarkers or reveal the mechanism of cisplatin-resistance in human ovarian cancers.

Screening of Domain-specific Target Proteins of Polo-like Kinase 1: Construction and Application of Centrosome/Kinetochore-specific Targeting Peptide

  • Ji, Jae-Hoon;Jang, Young-Joo
    • BMB Reports
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    • v.39 no.6
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    • pp.709-716
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    • 2006
  • Mammalian polo-like kinase 1 (Plk1) acts at various stages in early and late mitosis. Plk1 localizes at the centrosome and maintains this position through mitosis. Thereafter Plk1 moves to the kinetochore and midbody region, important sites during chromosome separation and cytokinesis. The catalytic domain of Plk1 is in the N-terminus region, whereas the non-catalytic region in the C-terminus of Plk1 has a conserved motif, named the Polobox. This motif is critical for Plk localization. EGFP proteins fused with the N-terminus and C-terminus of Plk1 localize in the nucleus and centrosomes, respectively. The core sequences of the polo-box (50 amino acids) also localize in Plk1 target organelles. To screen for domain-specific target proteins of Plk1, we constructed an N-terminal domain and a tandem repeat polo-box motif, and used them as templates in a yeast two-hybrid screen. The HeLa cell cDNA library indicated several proteins including the centrosome/kinetochore components or regulators, to be characterized as positive clones. Through in vitro protein binding analyses, we confirmed an interaction between these proteins and Plk1. The data reported from this study indicate that the N- and C- termini of Plk1 may function through recruitment and/or activation of domain-specific target proteins in dividing cells. Additionally, tandem repeats of the conserved core motif of the polo-box are sufficient for targeting and may be useful as a centrosome/kinetochore-specific targeting peptide.

Deastringent Peel Extracts of Persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) Protect Neuronal PC-12 and SH-SY5Y Cells against Oxidative Stress

  • Jeong, Da-Wool;Cho, Chi Heung;Lee, Jong Suk;Lee, Seung Hwan;Kim, Taewan;Kim, Dae-Ok
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1094-1104
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    • 2018
  • The peel of astringent persimmon (Diospyros kaki Thunb. cv. Cheongdo-Bansi) is a by-product of dried persimmon (gotgam). We investigated if deastringent peel extracts of persimmon cv. Cheongdo-Bansi had antioxidative and neuroprotective properties. Two different extracts were prepared: thermally and nonthermally treated persimmon peel extracts (TPE and NTPE, respectively). Both TPE and NTPE were fractionated sequentially in n-hexane, chloroform, ethyl acetate, n-butanol, and water. The TPE and NTPE ethyl acetate fractions had the highest total phenolic and flavonoid contents as well as antioxidant capacities among all the fractions. Pretreatment of neuronal PC-12 and SH-SY5Y cells with the TPE and NTPE ethyl acetate fractions increased cell viability after exposure to oxidative stress. The ethyl acetate fraction of TPE attenuated oxidative stress inside both PC-12 and SH-SY5Y cells more effectively than that of NTPE. Furthermore, the TPE and NTPE ethyl acetate fractions inhibited acetylcholinesterase and butyrylcholinesterase. Analysis of ultra-high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry results revealed gallic acid, kaempferol, kaempferol-3-O-galactoside, kaempferol-3-O-glucoside, quercetin, quercetin3-O-galactoside, quercetin-3-O-galactoside-2'-O-gallate, and quercetin-3-O-glucoside as the major phenolics of the TPE and NTPE ethyl acetate fractions. Taken together, these results suggest that the ethyl acetate fraction of deastringent persimmon peel is rich in antioxidants and has potential as a functional food to reduce oxidative stress.

Consensus channelome of dinoflagellates revealed by transcriptomic analysis sheds light on their physiology

  • Pozdnyakov, Ilya;Matantseva, Olga;Skarlato, Sergei
    • ALGAE
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    • v.36 no.4
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    • pp.315-326
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    • 2021
  • Ion channels are membrane protein complexes mediating passive ion flux across the cell membranes. Every organism has a certain set of ion channels that define its physiology. Dinoflagellates are ecologically important microorganisms characterized by effective physiological adaptability, which backs up their massive proliferations that often result in harmful blooms (red tides). In this study, we used a bioinformatics approach to identify homologs of known ion channels that belong to 36 ion channel families. We demonstrated that the versatility of the dinoflagellate physiology is underpinned by a high diversity of ion channels including homologs of animal and plant proteins, as well as channels unique to protists. The analysis of 27 transcriptomes allowed reconstructing a consensus ion channel repertoire (channelome) of dinoflagellates including the members of 31 ion channel families: inwardly-rectifying potassium channels, two-pore domain potassium channels, voltage-gated potassium channels (Kv), tandem Kv, cyclic nucleotide-binding domain-containing channels (CNBD), tandem CNBD, eukaryotic ionotropic glutamate receptors, large-conductance calcium-activated potassium channels, intermediate/small-conductance calcium-activated potassium channels, eukaryotic single-domain voltage-gated cation channels, transient receptor potential channels, two-pore domain calcium channels, four-domain voltage-gated cation channels, cation and anion Cys-loop receptors, small-conductivity mechanosensitive channels, large-conductivity mechanosensitive channels, voltage-gated proton channels, inositole-1,4,5-trisphosphate receptors, slow anion channels, aluminum-activated malate transporters and quick anion channels, mitochondrial calcium uniporters, voltage-dependent anion channels, vesicular chloride channels, ionotropic purinergic receptors, animal volage-insensitive cation channels, channelrhodopsins, bestrophins, voltage-gated chloride channels H+/Cl- exchangers, plant calcium-permeable mechanosensitive channels, and trimeric intracellular cation channels. Overall, dinoflagellates represent cells able to respond to physical and chemical stimuli utilizing a wide range of G-protein coupled receptors- and Ca2+-dependent signaling pathways. The applied approach not only shed light on the ion channel set in dinoflagellates, but also provided the information on possible molecular mechanisms underlying vital cellular processes dependent on the ion transport.

Effect of Heat and Moisture on the Phase Transition in Dimethylammonium-Facilitated CsPbI3 Perovskite (다이메틸암모늄 유도 CsPbI3 페로브스카이트 상의 상전이 거동에 대한 열과 수분의 영향)

  • Sohyun Kang;Seungmin Lee;Jun Hong Noh
    • Korean Journal of Materials Research
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    • v.33 no.8
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    • pp.344-351
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    • 2023
  • Cesium lead iodide (CsPbI3) with a bandgap of ~1.7 eV is an attractive material for use as a wide-gap perovskite in tandem perovskite solar cells due to its single halide component, which is capable of inhibiting halide segregation. However, phase transition into a photo inactive δ-CsPbI3 at room temperature significantly hinders performance and stability. Thus, maintaining the photo-active phase is a key challenge because it determines the reliability of the tandem device. The dimethylammonium (DMA)-facilitated CsPbI3, widely used to fabricate CsPbI3, exhibits different phase transition behaviors than pure CsPbI3. Here, we experimentally investigated the phase behavior of DMA-facilitated CsPbI3 when exposed to external factors, such as heat and moisture. In DMA-facilitated CsPbI3 films, the phase transition involving degradation was observed to begin at a temperature of 150 ℃ and a relative humidity of 65 %, which is presumed to be related to the sublimation of DMA. Forming a closed system to inhibit the sublimation of DMA significantly improved the phase transition under the same conditions. These results indicate that management of DMA is a crucial factor in maintaining the photo-active phase and implies that when employing DMA designs are necessary to ensure phase stability in DMA-facilitated CsPbI3 devices.

Efficient primary culture model of patient-derived tumor cells from colorectal cancer using a Rho-associated protein kinase inhibitor and feeder cells

  • HYE KYUNG HONG;DAE HEE PYO;TAE WON KIM;NAK HYEON YUN;YEO SONG LEE;SU JEONG SONG;WOO YONG LEE;YONG BEOM CHO
    • Oncology Letters
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    • v.42 no.5
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    • pp.2029-20238
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    • 2019
  • In vitro culture of patient-derived tumor cells offers many advantages in the development of novel therapies for colorectal cancer. Although various culture systems have been developed, the long-term expansion of patient-derived tumor cells remains challenging. The present results suggested that tumor cells isolated from colorectal cancer patient-derived xenografts can be efficiently immortalized in conditioned medium from irradiated feeder cells containing Y-27632, a rho-associated coiled-coil containing protein kinase (ROCK) inhibitor. Patient-derived tumor cells proliferated rapidly, reaching 90-95% confluence in ~6 days. Short tandem repeat analysis suggested that these tumor tissues and cultured cells presented 13 identical short tandem repeat loci, including Amelogenin, Penta E, Penta D, D2S1338 and D19S433. Their epithelial phenotype was confirmed by staining for epithelial cell adhesion molecule and cytokeratin 20, whereas vimentin was used as a mesenchymal marker. When cells were transferred to 3D cultures, they continued to proliferate, forming well-defined tumor spheroids. Expression levels of human telomerase reverse transcriptase and C-Myc mRNA were increased in cultured cells. Finally, immortalized cells were used for the screening of 65 anticancer drugs approved by the Food and Drug Administration, allowing the identification of gene-drug associations. In the present study, primary culture models of colorectal cancer were efficiently established using a ROCK inhibitor and feeder cells, and this approach could be used for personalized treatment strategies for patients with colorectal cancer.

Electrical and optical properties of back reflecting layer with AZO-Ag bilayer structure on a glass substrate for thin film Si solar cell applications (박막 Si태양전지 응용을 위한 유리기판 위의 AZO-Ag 이중구조 배면전극의 전기광학적 특성)

  • Park, Jaecheol;Hong, ChangWoo;Choi, YoungSung;Lee, JongHo;Kim, TaeWon
    • 한국신재생에너지학회:학술대회논문집
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    • 2011.05a
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    • pp.124.2-124.2
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    • 2011
  • 현재 박막형 태양전지는 실리콘계가 주류를 이루고 있으며, 유리기판 또는 유연성 기판에 비정질 실리콘 박막을 형성시킨 태양전지와 실리콘 기판 양면에 태양전지를 형성하는 방법 등 효율을 극대화시킨 이종접합 태양전지 등이 연구되고 있다. 예컨대 밴드갭이 서로 다른 박막들 간의 이종접합을 이용한 tandem 구조 및 triple 구조의 Si 박막 태양전지의 경우 13%대 변환효율을 나타낸다고 보고된 바 있다. 본 연구에서는 비정질 Si 박막 태양전지 내 흡수층의 효율을 최대화하기 위하여 AZO/Ag 이중구조 박막의 특성에 관한 연구를 수행하고자 한다. combinatorial sputtering system을 이용하여 AZO/Ag 이중구조 박막을 제작하였으며 타겟으로는 4-inch target(Ag, 2wt% Al2O3 doped ZnO)이 사용되었다. 유리기판 상에 combinatorial sputter system으로 상온에서 제작된 Ag 박막의 두께는 25nm로 성장시켰으며 연속공정으로 AZO 박막을 제작하였고, AZO 박막은 100~500nm의 두께경사를 나타내었다. 이 때 유리기판상에 성장된 Ag/AZO 박막의 면저항은 약 $2{\Omega}/{\Box}$ 값을 나타내었다. 본 발표에서는 AZO/Ag 이중 구조 박막의 우수한 전기적 특성을 기반으로 표면 거칠기 및 반사도 특성 등에 관하여 추가적으로 토론한다.

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Advances in Absorbers and Reflectors of Amorphous Silicon Oxide Thin Film Solar Cells for Tandem Devices (적층형 태양전지를 위한 비정질실리콘계 산화막 박막태양전지의 광흡수층 및 반사체 성능 향상 기술)

  • Kang, Dong-Won
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.30 no.2
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    • pp.115-118
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    • 2017
  • Highly photosensitive and wide bandgap amorphous silicon oxide (a-$SiO_x$:H) films were developed at low temperature ranges ($100{\sim}150^{\circ}C$) with employing plasma-enhanced chemical vapor deposition by optimizing $H_2/SiH_4$ gas ratio and $CO_2$ flow. Photosensitivity more than $10^5$ and wide bandgap (1.81~1.85 eV) properties were used for making the a-$SiO_x$:H thin film solar cells, which exhibited a high open circuit voltage of 0.987 V at the substrate temperature of $100^{\circ}C$. In addition, a power conversion efficiency of 6.87% for the cell could be improved up to 7.77% by employing a new n-type nc-$SiO_x$:H/ZnO:Al/Ag triple back-reflector that offers better short circuit currents in the thin film photovoltaic devices.

Recent Development of High-efficiency Silicon Heterojunction Technology Solar Cells (실리콘 이종접합 태양전지 개발동향)

  • Lee, Ahreum;Yoo, Jinsu;Park, Sungeun;Park, Joo Hyung;Ahn, Seungkyu;Cho, Jun-Sik
    • Current Photovoltaic Research
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    • v.9 no.4
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    • pp.111-122
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    • 2021
  • Silicon heterojunction technology (HJT) solar cells have received considerable attention due to advantages that include high efficiency over 26%, good performance in the real world environment, and easy application to bifacial power generation using symmetric device structure. Furthermore, ultra-highly efficient perovskite/c-Si tandem devices using the HJT bottom cells have been reported. In this paper, we discuss the unique feature of the HJT solar cells, the fabrication processes and the current status of technology development. We also investigate practical challenges and key technologies of the HJT solar cell manufacturers for reducing fabrication cost and increasing productivity.