• 제목/요약/키워드: Immobilization technique

검색결과 90건 처리시간 0.025초

Polyelectrolyte Micropatterning Using Agarose Plane Stamp and a Substrate Having Microscale Features on Its Surface

  • Lee, Min-Jung;Lee, Nae-Yoon;Lee, Sang-Kil;Park, Sung-Su;Kim, Youn-Sang
    • Bulletin of the Korean Chemical Society
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    • 제26권10호
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    • pp.1539-1542
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    • 2005
  • We have introduced polyelectrolyte micro-patterning technique employing agarose plane stamp and a hard substrate having microscale features on its surface. With this method, chemically micropatterned surfaces with both positive and negative functionalities were successfully embedded in well-defined microstructures, and selective impartment of charge functionalities was confirmed by patterning bead bearing surface charge. Furthermore, this technique allows highly sensitive immobilization of protein onto targeted surface simply by endowing functionalities, which extends the potential of its use as a tool for high-throughput protein microarray and proteomics. Because plane agarose stamp is free of structures on its surface, there is no concern for pattern collapse, and the combination of agarose plane stamp with patterned substrate is more suited for selective protein patterning compared with adopting surface-patterned agarose stamp with flat substrate. Our technique using agarose plane stamp and a substrate having microscale features on its surface suggests a range of possible applications, including the micropatterning of biofunctionalized copolymer having polyelectrolyte block, immobilization of micro- and nanoparticle with biofunctionalities such as biotin and streptavidine, and establishing optoelectronic microstructures with micro-beads on various surfaces.

Immobilization and Stability of Lipase from Mucor racemosus NRRL 3631

  • Adham, Nehad Zaki;Ahmed, Hanan Mostafa;Naim, Nadia
    • Journal of Microbiology and Biotechnology
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    • 제20권2호
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    • pp.332-339
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    • 2010
  • The lipase from Mucor racemosus NRRL 3631 was partially purified by fractional precipitation using 60% ammonium sulfate, which resulted in a 8.33-fold purification. The partially purified lipase was then immobilized using different immobilization techniques: physical adsorption, ionic binding, and entrapment. Entrapment in a 4% agar proved to be the most suitable technique (82% yield), as the immobilized lipase was more stable at acidic and alkaline pHs than the free enzyme, plus 100% of the original activity was retained owing to the thermal stability of the immobilized enzyme after heat treatment for 60 min at $45^{\circ}C$. The calculated half-lives (472.5, 433.12, and 268.5 min at 50, 55, and $60^{\circ}C$, respectively) and the activation energy (9.85 kcal/mol) for the immobilized enzyme were higher than those for the free enzyme. Under the selected conditions, the immobilized enzyme had a higher $K_m$ (11.11 mM) and lower $V_{max}$ (105.26 U/mg protein) when compared with the free enzyme (8.33 mM and 125.0 U/mg protein, respectively). The operational stability of the biocatalyst was tested for both the hydrolysis of triglycerides and esterification of fatty acids with glycerol. After 4 cycles, the immobilized lipase retained approximately 50% and 80% of its original activity in the hydrolysis and esterification reactions, respectively.

Development of Bioluminescence Immunoassay Using Photoprotein, Aequorin and Site-directed Immobilization

  • Shim, Yu-Nee;Rhee, In-sook
    • Bulletin of the Korean Chemical Society
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    • 제24권1호
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    • pp.70-74
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    • 2003
  • The heterogeneous bioluminescence immunoassay for digoxin was developed using photoprotein, native aequorin as a label and the site-directed immobilization technique based on avidin/biotin interaction. Aequorin is a bioluminescence protein, originally isolated from the jellyfish Aequoria Victoria and an attractive label in analytical applications because of sensitive detection due to virtually no background bioluminescent signal. Digoxin is a cardioactive drug, and its therapeutic level in serum is at low concentration with very narrow therapeutic index. The aequorin-digoxigenin conjugates were synthesized by the N-hydroxysuccinimide ester method and characterized in terms of bioluminescent residual activity. The resulting dose-response curve shows that the detection limit is $1.0\;{\times}\;10^{-10}\;M$ and a dynamic range is three orders of magnitude, which was obtained by $1.0\;{times}\;10^{-10}\;M$ conjugate and 0.9 μg/mL anti-digoxin antibody. Three structurally similar molecules to digoxin were examined for their cross-reactivity. None of these three compounds showed any crossreactivity with digoxin antibody employed in this study. Standard amounts of digoxin corresponding to the therapeutic range were spiked into the each serum solution. Study of the serum matrix effect indicated that correlation coefficient shows good agreement between luminescence light intensity between in buffer and in serum.

생물학적 펄프 파수처리를 위한 Phanerochaete chrnosporium의 고정화 방법 비교 (Comparison of ImmDbilization Techniques Using Phanerochaete chrysosporium for the Treatment of Pulp Waste Effluent)

  • 유인상
    • KSBB Journal
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    • 제8권4호
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    • pp.351-357
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    • 1993
  • 펄프 폐수처리를 위한 효과적인 생물학적 처리법 으로서 세 가지 고정화 방법과 자유균체를 이용한 방법을 사용하여 실험하였다. 균체로서는 Phaηe­r rochaete chrysosporium을 이용하였다. 고정화방법 중 Ca-alginate gel을 담체로 샤용한 방법이 가장 우수하였다. 이 방법은 안정성과 처리 효율변에서 좋은 결과를 보였으며, 약 400시간 동안 균체의 커다란 상태변화없이 반응시킬 수 있었다. 다른 고정화 방법들과 자유균체를 이용한 방법들 의 가장 큰 문제점은 생물 반응기 안에서의 공기 c channelling 현상에 의한 산소전달 저항과 균체끼리 의 clogging으로 인한 반응기 내부의 제어가 힘들게 된 것이었다.

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Biotransformation of Exogenous Monoterpenoids by Plant Cell Culture

  • Shin, Seung-Won
    • 생약학회지
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    • 제26권3호
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    • pp.227-238
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    • 1995
  • Recent reports on biotransformation of monoterpene alchols, aldehydes, acetates and epoxides are summerized. The studies have focused on stereospecific reaction of the functional groups of exogenous foreign substrates by foreign plant cells and micro-organisms. An another important aspect of research is the development of the immobilization technique for cells or related enzymes.

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Immobilization of enzymes on magnetic nanoparticles for biochip applications

  • Sohn, Ok-Jae;Rhee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
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    • pp.692-695
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    • 2003
  • In this work immobilization technique of enzymes onto the magnetic nanoparticles has been developed for biochip applications. Glucose oxidase and lactate dehydrogenase were immobilized on magnetic nanoparticles via cyanamide and glutaraldehyde. Immobilized enzymes had good operational and storage stability The immobilized glucose oxidase and lactate dehydrogenase were characterized by some factors(pH, temperature, and components of buffer solution etc) which affect the activity, In order to characterize the magnetic nanoparticles, we have used transmission electron microscopy(TEM), X-ray diffraction(XRD) and Fourier transform infrared(FTIR).

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Comparison of Two Different Immobilization Devices for Pelvic Region Radiotherapy in Tomotherapy

  • Kim, Dae Gun;Jung, James J;Cho, Kwang Hwan;Ryu, Mi Ryeong;Moon, Seong Kwon;Bae, Sun Hyun;Ahn, Jae Ouk;Jung, Jae Hong
    • 한국의학물리학회지:의학물리
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    • 제27권4호
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    • pp.250-257
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    • 2016
  • The purpose of this study was to compare the patient setup errors of two different immobilization devices (Feet Fix: FF and Leg Fix: LF) for pelvic region radiotherapy in Tomotherapy. Thirty six-patients previously treated with IMRT technique were selected, and divided into two groups based on applied immobilization devices (FF versus LF). We performed a retrospective clinical analysis including the mean, systematic, random variation, 3D-error, and calculated the planning target volume (PTV) margin. In addition, a rotational error (angles, $^{\circ}$) for each patient was analyzed using the automatic image registration. The 3D-errors for the FF and the LF groups were 3.70 mm and 4.26 mm, respectively; the LF group value was 15.1% higher than in the FF group. The treatment margin in the ML, SI, and AP directions were 5.23 mm (6.08 mm), 4.64 mm (6.29 mm), 5.83 mm (8.69 mm) in the FF group (and the LF group), respectively, that the FF group was lower than in the LF group. The percentage in treatment fractions for the FF group (ant the LF group) in greater than 5 mm at ML, SI, and AP direction was 1.7% (3.6%), 3.3% (10.7%), and 5.0% (16.1%), respectively. Two different immobilization devices were affected the patient setup errors due to different fixed location in low extremity. The radiotherapy for the pelvic region by Tomotherapy should be considering variation for the rotational angles including Yaw and Pitch direction that incorrect setup error during the treatment. In addition the choice of an appropriate immobilization device is important because an unalterable rotation angle affects the setup error.

고정기구의 사용이 골반부위 방사선조사영역의 변화에 미치는 오차분석 (The error analysis of field size variation in pelvis region by using immobilization device)

  • 김기환;강노현;김동욱;김준상;장지영;김용은;김재성;조문준
    • Journal of Radiation Protection and Research
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    • 제25권1호
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    • pp.31-36
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    • 2000
  • 방사선치료시 환자자세의 변화에 따라 방사선조사영역이 불일치하게 되어 주변의 정상조직에 불필요한 방사선조사가 생길수 있다. 본 연구에서는 골반부위 방사선 치료환자를 대상으로 Elecrtronic portal imaging device(EPID)로 고정기구를 사용한 방사선조사영역의 영상을 획득하여 고정기구사용에 따른 조사영역변화에 대한 오차를 분석하고자 한다 충남대학교병원 치료방사서과에서 1998년 8월부터 1999년 8월까지 21명의 골반부위 방사선치료환자를 대상으로 하였다. 환자들은 모두 앙와위자세를 유지하면서 치료를 받았고 고정기구를 사용하지 않은 11명과 고정기구를 사용한 10명으로 각각 나누어서 연구를 실행하였다 고정기구는 스터로폼을 이용하였다. 전체 치료과 정중 EPID를 이용하여 전면조사시 x(right-left)축, y(longitudinal)방향을, 측면조사는 z축(vertical) y축(longitudinal)방향을 대상으로 영상을 얻었다. 전체 치료과정중 얻은 영상들을 모의치료시 촬영한 기준필름과 matching기법을 이용하여 오차를 측정하였다. 고정기구를 사용하지 않은 11명 환자의 전면 조사영역의 경우 x축, y축으로의 평균이동값은 각각 0.19 mm, 0.48 mm이었고 계통이동과 우연이동은 평균이동으로부터 표준편차가 x축, y축으로 각각 2.38 mm, 2.19 mm와 1.92 mm, 1.29 mm이었다. 측면조사영역에서 z축, y축으혹의 평균이동값은 각각 -3.61 mm, 2.07 mm이었고 계통이동과 우연이동은 표준편차가 z축, y축므로 각각 3.20 mm, 2.29 mm와 2.73 mm, 1.62 mm이었다. 한편 고정기구를 사용란 10명 환자의 전면 조사영역에서 x축, y축으로의 평균이동값은 각각 0.71 mm, -1.07 mm이었고 계통이동과 우연이동은 표준편차가 x축, y축으로 각각 1.80 mm, 2.26 mm와 1.56 mm, 1.27 mm이었다. 측면조사영역에서 z축, y축으로의 평균이동값은 각각 -1.76 mm 1.08 mm 이었고 계통이동과 우연이동은 평균이동으로부터 표준편차가 x축, y축으로 각각 1.87 mm 2.83 mm와 1.68 mrn, 1.65 mm이었다. 본 연구에서는 고정기구를 사용한 환자군의 조사영역의 오차는 고정기구를 사용하지 않은 환자군의 조사영역의 오차에 비해 약간 감소됨을 확인할 수 있었다. 그리고 두그룹간의 우연오차는 감소하고 계통오차는 크게 변화하지 않는 경향을 보였다. 그러므로 골반부위 방사선 치료사 고정기구를 사용함으로써 방사선 조사영역에 대한 오차유발의 원인을 줄일 수 있을 것으로 생각된다.

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Flail Chest에서 Judet`s Struts를 이용한 수술치험 14례 (Treatment of flail chest with Judet`s strut)

  • 박병순;조용준;이동준
    • Journal of Chest Surgery
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    • 제24권4호
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    • pp.366-370
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    • 1991
  • This report describes our experience with the operative stabilization of flail chest with the use of Judet`s struts. In a series of 14 patients with flail chest, the method allowed shorter duration of artificial ventilation and decreased functional sequale. We find this technique to be better than previously published methods, since it provides better stabilization and immobilization of the ribs and thus obviates the need for artificial ventilation and prevents post-traumatic chest deformity.

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Volatile Organic Compound Specific Detection by Electrochemical Signals Using a Cell-Based Sensor

  • Chung, Sang-Gwi;Kim, Jo-Chun;Park, Chong-Ho;Ahn, Woong-Shick;Kim, Yong-Wan;Choi, Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • 제18권1호
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    • pp.145-152
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    • 2008
  • A cell-based in vitro exposure system was developed to determine whether oxidative stress plays a role in the cytotoxic effects of volatile organic compounds (VOCs) such as benzene, toluene, xylene, and chlorobenzene, using human epithelial HeLa cells. Thin films based on cysteine-terminated synthetic oligopeptides were fabricated for immobilization of the HeLa cells on a gold (Au) substrate. In addition, an immobilized cell-based sensor was applied to the electrochemical detection of the VOCs. Layer formation and immobilization of the cells were investigated with surface plasmon resonance (SPR), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). The adhered living cells were exposed to VOCs; this caused a change in the SPR angle and the VOC-specific electrochemical signal. In addition, VOC toxicity was found to correlate with the degree of nitric oxide (NO) generation and EIS. The primary reason for the marked increase in impedance was the change of aqueous electrolyte composition as a result of cell responses. The p53 and NF-${\kappa}B $ downregulation were closely related to the magnitude of growth inhibition associated with increasing concentrations of each VOC. Therefore, the proposed cell immobilization method, using a self-assembly technique and VOC-specific electrochemical signals, can be applied to construct a cell microarray for onsite VOC monitoring.