• 제목/요약/키워드: cell immobilization

검색결과 209건 처리시간 0.025초

동석물질에 대한 Photobacterium phosphoreum의 bioluminescence 특성 (The Character of Bioluminescence of Photobacterium phosphoreum with Toxic Substances)

  • 이홍주;이은수;박철진;이정건;전억한
    • KSBB Journal
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    • 제13권6호
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    • pp.693-699
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    • 1998
  • 고정화한 P. phosphoreum의 biolummescence악 독생물질과의 상관관계를 이용한 수계의 독성물질 momtonng 가능성을 조사하였다. P. phosphoreum을 이용한 수계의 독성물질 momtonng을 위한 적정 세포 농도는 최대 biolummescence intensnty를 보인 $O.D_{660}$ 1.0~1.2였으며 이 농도외 세포를 2.5% NaCl 용액에 희석하여 5.0%의 alginateo에 고정화 했을 때 biolummescence 유지도까 가장 좋았다. 이러한 방법으로 monitoring이 가능한 수계의 pH는 6.0~8.0이었다. Free cell과 고정화 세포로 $CdCl_2.H_20, PbCl_2, phenol $그리고 mtrophenol에 대한 bioluminescence를 조사하였을 때 고정화 방법에 의해 독성물질에 대한 민감성0] 증가되었다. 고정화 세포외 경우 각 독성 물질에 따라 20분 이내에 0.01~1.50ppm이하까지 mornitoring이 가능하였다. 뿐만 아니라 speciflC blolurninescence reclucLion rate, biolummescence mtenslLy의 ratio 그리고 Gamma value로 분석함으로서 독성물질의 농도와 biolummescence의 변화를 직선의 관계로 나타낼 수 있어 독성 물질의 농도 예측이 가능하다.

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Picomolar Scale Determination of Carbohydrates Covalently Immobilized on Activated Beads Using Hydroxyl Functionality

  • Yu, Jae-Hoon;Chun, Sung-Min;Park, Ho-Koon;Park, Yong-Keun;Jeong, Sun-Joo
    • BMB Reports
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    • 제32권1호
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    • pp.98-102
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    • 1999
  • Since carbohydrates are major mediators in cell-to-cell adhesion and communication, the development of specific and strong binders against them could generate promising therapeutics. As the first step towards that goal, sugar molecules have to be immobilized to be used as an affinity matrix. The amino functionality in sugar is the most active nucleophile for the immobilization, if the amino group is available. An alternative and general method is to use the hydroxyl group as a direct nucleophile, but the quantitation of immobilized hydroxyl groups is not easily done. To overcome this limitation, we have developed a method to immobilize various isomers of monosaccharides with p-nitrophenyl groups to the beads by using their hydroxyl groups. It was found that the amount of immobilized sugar was independent of the structure of the sugar, but was dependent on the number of hydroxyl groups. We also developed a sensitive method to quantify the amount of immobilized sugar at the picomolar scale by utilizing commercially available glycosidases to release a sensitive reporter molecule, p-nitrophenol, and detect it by HPLC. This new technique would allow a facile quantitation method for immobilized sugar molecules, which could be used as the affinity matrix to develop strong binders against biologically important sugars.

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Trigonopsis variabilis의 고정화 및 Cephalosporin C로부터 7$\beta$-(4-Carbohybutanamido)Cephalosporanic Acid의 전환 (Immobilization of Trigonopsis variabilis and Conversion of Cephalosporin C to 7$\beta$-(4-Caboxybutanamido)Cephalosporanic Acid)

  • 김종균;임재윤
    • 한국미생물·생명공학회지
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    • 제22권3호
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    • pp.296-303
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    • 1994
  • An immobilized Trigonopsis variabilis cells having an high activity of D-amino acid oxidase(DAO) was used to convert CPC into GL-7-ACA. The optimal pH of the reaction system was 8.0-8.5, and the optimal temperature was 40$\circ$C. When immobilized cell was used repeatedly in semi-batchwise reaction, the system retained 80% of the initial activity after used of 12 times for over 12 hours. The storage stability of the immobilized cell was maintained for 30 days at 4$\circ$C. The CPC concentration for the maximal reaction rate was about 30 mM and 40 mM for free and immobilized cells, respectively. Substrate inhibition of CPC concentration more than 50 mM was overcomed by 20~25% by immobilization. Pure oxygen supply into reaction system was most efficient in D-amino acid oxidase reaction. Continuous conversion to GL-7-ACA from CPC has been developed with an bioreactor system containing immobilized T variabilis cells. By opera- tion of the reactor for 5 hours, the average conversion yield of >80% and GL-7-ACA production of 40~45 mM per hour could be obtained.

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쥐의 비복근 섬유의 변화에 대한 형태학적 연구 (A Morphological Study on the Changes in Rat's Gastrocnemius)

  • 허양훈;최재청
    • The Journal of Korean Physical Therapy
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    • 제10권2호
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    • pp.71-76
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    • 1998
  • 본 연구는 실험용 려 12마리를 정상관, 2주 고정군, 4주 고정군에 각각 4마리씩 배정하고 석고 고정 기간 후 비복근의 절편을 취하여 투과전자현미경으로 형태학적 관찰을 한 결과를 다음과 같이 요약할 수 있다. 골격근을 짧은 위치로 일정 기간 고정하면 근절의 길이와 폭은 물론 삼조체, 글리코겐, 필라멘트 단백질, 미토콘드리아와 근 세포 핵의 모양까지도 심한 변화가 일어나기 때문에 구축과 위축의 개념에 근원 섬유의 조직, 형태학적 의의를 추가하여야 할 것으로 사료된다.

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Aspergillus Phoenicis Whole Cell의 ${\beta}-Galactosidase$ 활성(活性)에 관한 연구(硏究) (Studies on the ${\beta}-Galactosidase$ Activity of Whole Cell Aspergillus Phoenicis)

  • 김말남
    • 한국균학회지
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    • 제11권3호
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    • pp.109-114
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    • 1983
  • Aspergillus phoenicis의 ${\beta}-galactosidase$ 활성을 ONPG와 lactose를 기질로서 사용하여 조사하였다. 이 효소 활성은 성장의 대수기 동안은 서서히 증가하였으나 정지기가 시작되면서 급격하게 감소하였다. 또한 이 효소는 높은 온도에서도 좋은 효소 활성을 유지하였으며, 산성 pH에서 최고의 효소활성을 나타내었다. ${\beta}-galactosidase$는 lactose보다 ONPG에 대한 기질의 친화력이 더 좋았으며, 효소 활성도 ONPG의 경우가 더 높았다. Lactose의 가수분해율은 반응액중의 lactose의 농도가 낮을 수록 높았으며, 사용균의 무게에 따라 초기에는 증가하다가 어느 수준 이상에서 부터는 점근값을 나타내었다. 효소의 활성은 효소의 고정 방법 및 조건에 영향을 받았으며, matrix의 가교가 pH 7.2 및 0.35 vol. %의 glutaraldehyde 농도에서 행하여졌을 때, 가장 높은 효소 활성을 보였다.

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Bio-functionalization of the Single Layer Graphene for Detecting the Cancer Cell

  • Oh, Hyung Sik;Park, Wanjun
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2014년도 제46회 동계 정기학술대회 초록집
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    • pp.429.1-429.1
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    • 2014
  • We present a method of surface functionalization of a single layer graphene for linking and detecting MDA-MB-231 human breast cancer cell. The methodology is done by utilizing 1-pyrenebutanoic acid and succinimidyl ester for immobiling CD44 antibodies. This work shows that the single layer graphene is an efficient fixing substance to capture the MDA-MB-231 human breast cancer cell, selectively. The immobilization method of the cancer cell on the graphene layer will be an effective cell counting system. Moreover usage of the linking with non-covalent bonding is expected to develope a sensor scheme of electrical cell-detecting diagnosis system.

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이중실관 반응기에서 E. coli의 고농도 배양 (High Density Cell Cultivation of Escherichia coli in a Dual Hollow Fiber Bioreactor)

  • Chung, Bong-Hyun;Chang, Ho-Nam;Kim, In-Ho
    • 한국미생물·생명공학회지
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    • 제13권3호
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    • pp.209-212
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    • 1985
  • 산소 투과율이 좋은 silicone tube안쪽에 nutrient 공급을 위한 3개의 isotropic polypropylene hollow fiber 3개를 넣어 제작된 이중 실관 반응기에서 E. coli cell을 immobilization 하여 cell density와 packing characteristics를 조사해 보았다. E. coli cell들은 거의 100% Packing되어 동물조직에서 처럼 층을 이루면서 자랐고 ceil density를 측정해 본 결과 약 550g/$\ell$고농도 세포배양이 가능하였다.

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보심건비탕(補心健脾湯) 투여가 Stress 유발 Mouse의 Hypothalamus 유전자 발현에 미치는 영향 (Effects of Boshimgeonbi-tang on Gene Expression in Hypothalamus of Immobilization-stressed Mouse)

  • 이승희;장규태;김장현
    • 동의생리병리학회지
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    • 제19권6호
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    • pp.1585-1593
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    • 2005
  • The genetic effects of restraint stress challenge on HPA axis and the therapeutic effect of Boshimgeonbi-tang on the stress were studied with cDNA microarray analyses, RT-PCR on hypothalamus using an immobilization-stress mice as an animal model. Male CD-1 mice were restrained in a tightly fitted and ventilated vinyl holder for 2hrs once a day, and this challenge was repeated for seven· consecutive days. In the change of body weight it showed that the Boshimgeonbi-tang is effected recovery on weight loss caused by the immobilization-stress. Seven days later, total RNA was extracted from the organs of the mouse, body-labeled with $CyDye^{TM}$ fluorescence dyes and then hybridized to CDNA microarray chip. Scanning and analyzing the array slides were carried out using GenePix4000 series scanner and GenePix $Pro^{TM}$ analyzing program, respectively. The expression profiles of 109 genes out of 6000 genes on the chip were significantly modulated in hypothalamus by the immobilization stress. Energy metabolism-, lipid metabolism-, apoptosis-, stress protein, transcriptional factor, and signal transduction-related genes were transcriptionally activated whereas DNA repair-, protein biosysthesis-, and structure integrity-related genes were down-regulated in hypothalamus. The 58 genes were up-regulated by the mRNA expression folds of 1.5 to 7.9. and the 51 genes were down-regulated by 1.5 - 5.5 fold. The 11 genes among them were selected to confirm the expression profiles by RT-PCR. The mRNA expression levels of Tnfrsf1a (apoptosis), Calm2 (cell cycle), Bag3 (apoptosis), Ogg1 (DNA repair), Aatk (apoptosis), Dffa (apoptosis), Fkbp5 (protein folding) were restored to the normal one by the treatment of Boshimgeonbi-tang.

Optimization of Catechol Production Using Immobilized Resting Cells of Pseudomonas putida in Aqueous/organic Two-phase System

  • Chae, Hee-Jeong;Yoo, Young-Je
    • Journal of Microbiology and Biotechnology
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    • 제7권5호
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    • pp.345-351
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    • 1997
  • An aqueous/organic two-phase reaction system was applied to the production of catechol using immobilized resting cells of Pseudomonas putida CY 400. Water/ethyl ether system was used because of high partition coefficient of catechol and thus to reduce the product inhibition and degradation. Among the tested immobilization carriers, polyacrylamide gel gave the highest catechol productivity. The immobilization seemed to protect the cells against solvent toxicity. From the simulation of reaction conditions based on two-phase models, it was found that there was an optimum acetate concentration at fixed benzoate and cell concentrations for the catechol productivity. A lower phase volume ratio (lower fraction of organic phase) gave a higher productivity. However, the substrate conversion was low at low phase volume ratio.

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Preparation and Atomic Force Microscopy (AFM) Characterization of DNA Scaffolds as a Template for Protein Immobilization

  • Kim, Hyeran;Lee, Hyun Uk;Lee, Jouhahn
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2014년도 제46회 동계 정기학술대회 초록집
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    • pp.411.2-411.2
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    • 2014
  • The design of DNA nanostructures is of fundamental importance, the intrinsic value of DNA as a building-block material lies in its ability to organize other bio-molecules with nanometer-scale spacing. Here, we report the fabrication of DNA scaffolds with nano-pores (<10 nm size) that formed easily without the use of additives (i.e., avidin, biotin, polyamine, or inorganic materials) into large-scale structures by assembling DNA molecules at near room temperature ($30^{\circ}C$) and low pH (~5.5). Protein immobilization results also confirmed that a fibronectin (FN) proteins/large scale DNA scaffolds/aminopropylytriethoxysilane (APS)/SiO2/Si substrate with high sensitivity formed in a well-defined manner. The DNA scaffolds can be applied for use with DNA-based biochips, biophysics, and cell biology.

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