• 제목/요약/키워드: immobilized bead

검색결과 132건 처리시간 0.026초

Characteristics of IEF Patterns and SDS-PAGE Results of Korean EPO Biosimilars

  • Kang, Min-Jung;Shin, Sang-Mi;Yoo, Hey-Hyun;Kwon, Oh-Seung;Jin, Chang-Bae
    • Bulletin of the Korean Chemical Society
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    • 제31권9호
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    • pp.2493-2496
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    • 2010
  • Erythropoietin (EPO) is mainly produced in kidney and stimulates erythropoiesis. The use of recombinant EPOs for doping is prohibited because of its performance enhancing effect. This study investigated whether biosimilar EPOs could be differentiated from endogenous one by iso-electro-focusing plus double blotting and SDS-PAGE for antidoping analysis. The established method was validated with positive control urine. The band patterns were reproducible and meet the criteria, which was made by world anti doping agency (WADA). Isoelectric focusing was conducted in pH range 2 to 6. Recormon (La Roche), Aropotin (Kunwha), Epokine (CJ Pharm Co.), Eporon (Dong-A), Espogen (LG Life Sciences), and Dynepo (Shire Pharmaceuticals) were detected in basic region. All biosimilars showed discriminative isoelectric profiles from endogenous EPO profiles, but they showed different band patterns with the reference one except Epokine (CJ Pharm Co.). Next, SDS-PAGE of biosimilar EPOs resulted in different molecular weight patterns which were distributed higher than endogenous EPO. Commercial immune assay kit as an immune affinity purification tool and immobilized antibody coated magnetic bead were tested for the purification and concentration of EPO from urinary matrix. The antibody-coated magnetic bead gave better purification yield. The IEF plus double blotting and SDS-PAGE with immunoaffinity purification method established can be used to discriminate biosimilar EPOs from endogenous EPO.

연속 생물반응기 안에서 유출 발효에 의한 알코올 생산 (Alcohol Production by Extractive Fermentation in a Continuous Bioreactor)

  • 김재형;전순배이기영김동운
    • KSBB Journal
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    • 제4권1호
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    • pp.21-30
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    • 1989
  • Lauryl alcohol을 추출제로 사용한 에탄올 추출발효를 통하여 다음과 같은 결론을 얻을 수 있었다. 1. GC를 이용한 분배계수의 측정은 Fredenslund의 UNIFAC법을 이용한 계산치보다는 고천 낮은 값을 보이나 Minier의 측정치와는 잘 일치하고, 에탄올 농도 2-10%일때 분배계수는 0.6에 달했다. 2. 물과 에탄올은 비점이 비슷하여 95% 이상의 고순도 에탄올을 단증류로 얻기 어렵지만, LaOH는 비점($225^{\circ}C$)이 높아 에탄올과 비점차가 많이 나므로 대기압하에서 단 한번의 단증류로 87% 이상의 에탄올을 거의 모두 회수할 수 있었다. 3. TBP의 경우와 마찬가지로, LaOH의 농도가 높아 질수록 lag phase는 연장도지만, 기질 소모속 도는 추출제가 없는 경우와 비교했을때 크게 변하지 않았으며, 적응과정을 거침으로써 LaOH에 의한 lag phase 지연호과가 제거되었고, 고정화 방법 또한 lag phase를 줄이는데 교과적인 방법이었다. 4. $400\;g/\;{\ell}$ 포도당 용액의 회분 추출발생에서 LaOH/medium의 비가 4였을때 포도당은 거의 전화가 되었으며, 전체에탄올생산성은 $2.75g/\;{\ell}.hr$로 비유출의 $0.99g/\;{\ell}{\cdot}hr$보다 2.78배 증가하였다. 5. 고정화 연속 혼합 발효조에서 기질유량 $25\;{m{\ell}}/hr$, LaOH유량 $100\;{m{\ell}}/hr$일때 생산성은 $5.23\;g/{\ell}.hr$로 비유출의 $3.06\;g/{\ell}.hr$보다 1.7배 증배하였다. 6. film fermentor에서 기질농도 $200\;g/{\ell}$, 기질유출 $25\;{m{\ell}}/hr$, LaOh 유량 $100\;{m{\ell}}/hr$일때 생산성이 $5.01\;g/{\ell}.hr$로 연속 혼합 발효조에서 보다 낮은 값을 보였으나, 대체로 비슷한 양상을 보였으며, 비에탄올생산성은 고천 더 놓은 값을 나타내었고, 연속 혼합 발효조 안의 bead 내 효모보다 낮은 전단력을 받기 때문에 세포가 덜 유출되었다.

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In vitro functional assenssment of bioartificial liver system using immobilized porcine hepatocyte spheroids

  • Lee, Ji-Hyun;Lee, Doo-Hoon;Yoon, Hee-Hoon;Jung, Doo-Hee;Park, Jung-Keug;Kim, Sung-Koo;Lee, Kwang-Woong;Lee, Suk-Koo
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XIII)
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    • pp.305-306
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    • 2003
  • To treat fulminant hepatic failure (FHF) patients, various extracorporeal bioartificial liver (BAL) systems have been developed. Several requirements should be met for the development of BAL systems: hepatocytes should be cultured in a sufficiently high density; their metabolic functions should be of a sufficiently high level and duration; and the BAL systems module should permit scaling-up and aseptic handling. Several investigators have found that freshly isolated primary hepatocytes can be cultured into three dimensional, tightly packed, freely suspended, multicellular aggregates, or spheroids. These specialized cell structures exhibited enhanced liver specific functions and a prolonged differentiated state compared to cells maintained in a monolayer culture. Cells in spheroids appear to mimic the morphology and ultrastructure of the in vivo liver lobule. The ability of hepatocytes to organize into three-dimensional structures was hypothesized to contribute to their enhanced liver-specific activities. In this study, the ammonia removal rate and urea secretion rate of pig hepatocytes spheroids encapsulated in Ca-alginate bead were determined. A packed-bed bioreactor with encapsulated pig hepatocytes was devised as BAL support system. The efficacy of the system was evaluated in vitro.

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Al(OH)3를 PVC로 고정화한 PVC-Al(OH)3 비드의 제조와 수중의 불소 이온의 흡착 특성 (Preparation of PVC-Al(OH)3 Beads Immobilized Al(OH)3 with PVC and their Adsorption Characteristics for Fluoride Ions from Aqueous Solution)

  • 유해나;감상규;이민규
    • 한국환경과학회지
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    • 제23권5호
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    • pp.887-893
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    • 2014
  • In order to remove fluoride ions from aqueous solution, PVC-$Al(OH)_3$ beads were prepared by immobilizing $Al(OH)_3$ with polyvinyl chloride (PVC). The prepared PVC-$Al(OH)_3$ bead was characterized by using SEM, EDS and Zeta potential. Dependences of pH, contact time and initial fluoride concentration on the adsorption of fluoride ions were studied. The optimal pH was in the range of 4~10. The adsorption was rapid during the initial 12 hr, and equilibrium was attained within 72 hr. The adsorption rate of fluoride ions by PVC-$Al(OH)_3$ beads obeyed the pseudo-second-order kinetic model. The maximum adsorption capacity obtained from Langmuir isotherm model was found to be 62.68 mg/g.

비드를 이용한 면역분석용 마이크로필터 칩의 제작 (Microfilter Chip Fabrication for Bead-Based Immunoassay)

  • 이승우;안유민;채영규
    • 대한기계학회논문집A
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    • 제28권9호
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    • pp.1429-1434
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    • 2004
  • Immunoassay is one of the important analytical methods for clinical diagnoses and biochemical studies, but needs a long time, troublesome procedures and expensive reagents. In this study, therefore, we propose the micro filter chip with microbeads for immunoassay, which has pillar structures. The advantage of the proposed micro filter chip is to use simple fabrication process and cheap materials. The mold was made by the photolithography technique with Si wafer and negative photoresist SU-8. The replica was made of PDMS, bonded on the pyrex glass. The micro filter chip consists of inlet channel, filter chamber and outlet channel. HBV (Hepatitius B virus) monoclonal antibody (Ag1) labeled with biotin were immobilized onto streptavidin coated beads of 30∼50 $\mu$m size. Fluorescein isothiocyanate (FITC)-labeled HBV monoclonal antibody (Ag8) was used to detect HBsAg (Hebatitis B virus surface Antigen), and fluorescence intensity was monitored by epi-fluorescence microscope. In this study, the immune response of less than 30 min was obtained with with the use of 100 $m\ell$ of sample.

Immobilization of Penicillium citrinum by Entrapping Cells in Calcium Alginate for the Production of Neo-Fructooligosaccharides

  • Lim, Jung-Soo;Park, Seung-Won;Lee, Jin-Won;Oh, Kyeong-Keon;Kim, Seung-Wook
    • Journal of Microbiology and Biotechnology
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    • 제15권6호
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    • pp.1317-1322
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    • 2005
  • This work describes neo-fructooligosaccharides (neo-FOSs) production using the immobilized mycelia of Penicillium citrinum. Some critical factors were evaluated to optimize maximal production of neo-FOS. Optimal alginate and cell concentrations were determined to be $1.96\%$ alginate and $7.17\%$ cell, respectively, by statistical analysis. The optimal concentration of $CaCl_{2}$, which is related to bead stability, was determined to be 2 M. It was possible to increase the neo-FOS production by adding 15 units of glucose oxidase to the batch reaction. By co-immobilizing cells and glucose oxidase, neoFOS productivity increased $123\%$ compared with the whole-cell immobilization process. Based on the results above, a co-immobilization technique was developed and it can be utilized for large-scale production.

Dioxane을 용매로 한 PVC-LMO 비드의 제조와 Li+ 흡착특성 (Preparation of PVC-LMO Bead with Dioxane Solvent and Adsorptive Properties of Lithium Ions)

  • 감상규;유해나;이민규
    • 한국환경과학회지
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    • 제23권7호
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    • pp.1289-1297
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    • 2014
  • In this study, PVC-LMO beads were prepared by immobilizing lithium manganese oxide (LMO) with poly vinyl chloride (PVC) diluted in dioxane solvent. XRD and SEM analysis confirmed that LMO was immobilized well in PVC-LMO beads. The diameter of PVC-LMO beads prepared by dioxane solvent was about 2 mm. The adsorption experiments of lithium ions by PVC-LMO beads were conducted batchwise. The optimum pH was pH 10. The adsorption characteristics of lithium ions by PVC-LMO beads was well described by the pseudo-second-order kinetic model. The maximum adsorption capacity obtained from Langmuir model was 24.25 mg/g. The thermodynamic parameters such as ${\Delta}H^{\circ}$, ${\Delta}S^{\circ}$ and ${\Delta}G^{\circ}$ were evaluated. The calculated ${\Delta}G^{\circ}$ was between -6.16 and -4.14 kJ/mol (below zero), indicating the spontaneous nature of $Li^+$ adsorption on PVC-LMO beads. Also, the results showed that PVC-LMO beads prepared in this study could be used for the removal of lithium ions from seawater containing coexisting ions such as $Na^+$, $K^+$, $Mg^{2+}$ and $Ca^{2+}$.

Micro Pre-concentration and Separation of Metal Ions Using Microchip Column Packed with Magnetic Particles Immobilized by Aminobenzyl Ethylenediaminetetraacetic Acid

  • Kim, Y.H.;Kim, G.Y.;Lim, H.B.
    • Bulletin of the Korean Chemical Society
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    • 제31권4호
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    • pp.905-909
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    • 2010
  • Magnetic beads (Dynabeads$^{(R)}$) embedded in ~1 micron size polystyrene beads bearing surface carboxylic acid groups were modified with aminobenzyl ethylenediaminetetraacetic acid (ABEDTA) to concentrate or separate metal ions using pH gradients on micro and nano scales. The immobilization of ABEDTA was achieved by amide formation. The presence of the metal chelating functional group in the fully deprotonated form was confirmed by FT-IR. The chelation efficiency of beads was tested by determining metal ions in supernatant using GFAAS when pH gradients from 3 to 7. Mixtures of Cu and Mg and of Cd and Mn (at 10 ng/mL of metal) were separated as the difference in formation constant with the functional group of ABEDTA. The separation was repeated twice with relative standard deviation of <18%. A polydimethylsiloxane (PDMS) microchip column packed with EDTA-coated magnetic beads was optimized to concentrate metal ion for practical applications by eluting a Cu solution of micro scale at pH 3.

제주 화산석으로 합성한 제올라이트를 Polyacrylonitrile에 고정화한 흡착제를 이용한 구리와 스트론튬 이온의 제거 (Removal of Cu and Sr Ions using Adsorbent Obtained by Immobilizing Zeolite Synthesized from Jeju Volcanic Rocks in Polyacrylonitrile)

  • 이창한;이민규
    • 한국환경과학회지
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    • 제27권12호
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    • pp.1215-1226
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    • 2018
  • In this study, PAN-SZ (polyacrylonitrile scoria zeolite) beads were prepared by immobilizing Na-A zeolite (SZ-A) synthesized from Jeju volcanic rocks (scoria) on the polymer PAN. FT-IR and TGA analysis results confirmed that the SZ-A was immobilized in the PAN-SZ beads. SEM images showed that the PAN-SZ beads are a spherical shape with 2 mm diameter and exhibit a porous inner structure inside the bead. The most suitable mixing ratio of PAN to SZ-A as the adsorbent for removing Sr ions was PAN/SZ-A = 0.2 g/0.3 g. The adsorption kinetic data for Cu and Sr ions were fitted well with the pseudo-second-order model. The Cu and Sr ion uptakes followed a Langmuir isotherm model and the maximum adsorption capacities at $20^{\circ}C$ were 84.03 mg/g and 75.19 mg/g, respectively. The amount of Sr ion adsorbed by SZ-A on the PAN-SZ beads was about 160 mg/g, which was similar to that adsorbed by SZ-A powder. Thus, the PAN-SZ beads prepared in this study are considered to be effective adsorbents for removing metal ions in aqueous solutions.

화학.생물학적 황화수소 제거 공정에 있어서 고정화 세포를 이용한 철산화 속도 증진 (Enhancement of Iron Oxidation Rate by Immobilized Cells in Chemo-biological Process for $H_2S$ Removal)

  • 김태완;김창준;장용근
    • KSBB Journal
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    • 제14권5호
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    • pp.585-592
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    • 1999
  • 본 연구는 황화수소 가스 제거를 위한 화학 . 생물학적 복합공정의 전체 효율을 결정하는 생물반응기에 의한 Fe(II) 산화 속도를 증진시키고자 배지 최적화와 함께 고정화 세포 반응기 시스템을 개발하는 것을 목표로 하였다. 고정화 담체로는 celite beads를 선정하였고 반응기는 airlift type의 반응기를 사용하였다. 먼저, 철침전물(jarosite) 최소화 배지 개발을 위한 연구를 수행하였고, 그 결과 기존에 사용되어졌던 9K 배지 사용시 질소원 및 인 공급원으로 사용된 $(NH_4)_2SO_4$$K_2HPO_4$가 제외되고 $(NH_4)_2HPO_4$가 대체된 M16 배지를 사용하였을 때 Fe(II) 산화 속도의 감소가 없음은 물론 jarosite 생성이 거의 없음을 확인할 수 있었다. 또한 M16 배지의 초기 pH 변화에 따른 철 산화 거동 및 jarosite 생성 측면을 조사한 결과 초기 pH 1.8이 최적임을 확인할 수 있었다. 다음으로 celite beads에 세포를 고정화 한 후 jarosite 생성 최소화 배지(M16 배지)에서의 고정화 세포에 의한 철 산화 거동을 고찰하였다. 반복 회분식 배양 결과, 회분식 배양의 결과 최대 Fe(II) 산화 속도에 이르러서는 거의 일정해지는 결과를 보였다. 반분회분식 배양의 결과 최대 Fe(II) 산화 속도가 2.33 g/L . h이었다. 연속 조업을 수행한 결과 현탁 세포의 최대 비성장 속도보다 높은 희석속도에서 최대 Fe(II) 산화 속도가 결정되었다. 최대 Fe(II) 산화속도는 2.14 g/L . h이었으며, 이??의 희석속도는 0.25 $h^{-1}$이었다. 반복 회분식 및 연속 조업기간 동안 Fe(total)의 농도는 초기 Fe(II) 농도와 거의 비슷하게 유지되었고 이러한 사실로부터 jarosite가 거의 생성되지 않았음을 확인하였다.

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