• Title/Summary/Keyword: affinity separation

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Selective adsorption of Ba2+ using chemically modified alginate beads with enhanced Ba2+ affinity and its application to 131Cs production

  • Kim, Jin-Hee;Lee, Seung-Kon
    • Nuclear Engineering and Technology
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    • v.54 no.8
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    • pp.3017-3026
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    • 2022
  • The 131Cs radioisotope with a short half-life time and high average radiation energy can treat the cancer effectively in prostate brachytherapy. The typical 131Cs production processes have a separation step of the cesium from 131Ba to obtain a high specific radioactivity. Herein, we suggested a novel 131Cs separation method based on the Ba2+ adsorption of alginate beads. It is necessary to reduce the affinity of alginate beads to cesium ions for a high production yield. The carboxyl group of the alginate beads was replaced by a sulfonate group to reduce the cesium affinity while reinforcing their affinity to barium ions. The modified beads exhibited superior Ba2+ adsorption performances to native beads. In the fixed-bed column tests, the saturation time and adsorption capacity could be estimated with the Yoon-Nelson model in various injection flow rates and initial concentrations. In terms of the Cs elution, the modified alginate showed better performance (i.e., an elution over 88%) than the native alginate (i.e., an elution below 10%), indicating that the functional group modification was effective in reducing the affinity to cesium ions. Therefore, the separation of cesium from the barium using the modified alginate is expected to be an additional option to produce 131Cs.

Basic study on high gradient magnetic separation of nano beads using superconducting magnet for antibody purification

  • Jeongtae Kim;Insung Park;Gwantae Kim;Myunghwan Sohn;Sanghoon Lee;Arim Byun;Jin-sil Choi;Taekyu Kim;Hongsoo Ha
    • Progress in Superconductivity and Cryogenics
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    • v.25 no.4
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    • pp.60-64
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    • 2023
  • The manufacturing process of antibody drugs comprises two main stages: the upstream process for antibody cultivation and the downstream process for antibody extraction. The domestic bio industry has excellent technology for the upstream process. However, it relies on the technology of foreign countries to execute downstream process such as affinity chromatography. Furthermore, there are no domestic companies capable of producing the equipment for affinity chromatography. High gradient magnetic separation technology using a high temperature superconducting magnet as a novel antibody separation and purification technology is introduced to substitute for the traditional technology of affinity chromatography. A specially designed magnetic filter was equipped in the bore of the superconducting magnet enabling the continuous magnetic separation of nano-sized paramagnetic beads that can be used as affinity magnetic nano beads for antibodies. To optimize the magnetic filter that captures superparamagnetic nanoparticles effectively, various shapes and materials were examined for the magnetic filter. The result of magnetic separation experiments show that the maximum separation and recovery ratio of superparamagnetic nanoparticles are 99.2 %, and 99.07 %, respectively under magnetic field (3 T) and flow rate (600 litter/hr).

Affinity chromatography and capillary electrophoresis for analysis of the yeast ribosomal proteins

  • Goyder, Miriam S.;Willison, Keith R.;Klug, David R.;DeMello, Andrew J.;Ces, Oscar
    • BMB Reports
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    • v.45 no.4
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    • pp.233-238
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    • 2012
  • We present a top down separation platform for yeast ribosomal proteins using affinity chromatography and capillary electrophoresis which is designed to allow deposition of proteins onto a substrate. FLAG tagged ribosomes were affinity purified, and rRNA acid precipitation was performed on the ribosomes followed by capillary electrophoresis to separate the ribosomal proteins. Over 26 peaks were detected with excellent reproducibility (<0.5% RSD migration time). This is the first reported separation of eukaryotic ribosomal proteins using capillary electrophoresis. The two stages in this workflow, affinity chromatography and capillary electrophoresis, share the advantages that they are fast, flexible and have small sample requirements in comparison to more commonly used techniques. This method is a remarkably quick route from cell to separation that has the potential to be coupled to high throughput readout platforms for studies of the ribosomal proteome.

The Relationship between Affinity of Membrane and Optimum Operation Conditions in the Pervaporation of Aqueous Ethanol (에탄올 수용액의 투과증발에 있어서 막의 친화성과 최적 조업조건의 관계)

  • 전종기;명완재;임선기
    • Membrane Journal
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    • v.1 no.1
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    • pp.34-43
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    • 1991
  • The relationships between affinity of membranes and optimum operation conditions were investigated in the pervaporation of water(1)/ethanol(2) mixture through cellulose acetate(CA) membranes having more affinity to water and silicone rubber(SR) membranes having more affinity to ethanol. CA and SR membranes were prepared and amount of sorption, sorption selectivity, pervaporation separation factor and pervaporation rate in both of membranes were determined and compared. The effects of downstream pressure were analyzed using Thompson diagram and the sorption and pervaporation characteristics with composition of feed and operation temperature were examined in terms of affinity, activity coefficient, plasticizing effect and activation energy of individual species. In the separation of water through CA membranes, high performance of both pervaporation separation factor (water to ethanol, $[\alpha^2_1]_{PV}$) and pervaporation rate was obtained in the conditions of low downstream pressure, middle range of feed concentration and high temperature. In the separation of ethanol through SR membranes, pervaporation separation factor(ethanol to water, $[\alpha^2_1]_{PV}$) increased with downstream pressure and decreased with concentration of ethanol in feed and operation temperature, while pervaporation rate showed opposite trends to those of ($[\alpha^2_1]_{PV}$).

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Affinity Filtration Chromatography of Proteins by Chitosan and Chitin Membranes: 2. Separation of BSA and Lysozyme (키토산 및 키틴 막에 의한 단백질의 친화 여과 크로마토그래피: 2. BSA 및 Lysozyme의 분리)

  • Youm, Kyung-Ho;Yuk, Yeong-Jae
    • Membrane Journal
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    • v.19 no.2
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    • pp.113-121
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    • 2009
  • Porous affinity chitosan and chitin membranes with good mechanical strength and high protein binding capacity were prepared by using silica particles as porogen. The maximum binding capacity of affinity chitosan membrane for BSA protein is 21.8mg/mL, and that of affinity chitin membrane for lysozyme enzyme is 26.1mg/mL. Chromatographic separations of BSA and lysozyme proteins using the porous affinity chitosan and chitin membranes were performed with change of the flow rate, loading amount and concentration of protein loading solutions. Protein eluted amount and binding yield were calculated from the filtration chromatograms consisted of loading/washing/elution sequences. Protein binding amount and yield were increased with decreasing of flow rate, increasing of loading amount and concentration of protein loading solutions. Those results suggest that the porous chitosan and chitin membranes prepared by using silica particles as porogen are suitable in affinity filtration chromatography for large scale separation of proteins.

Chromatography separation of proteins by macroporous chitosan and chitin affinity membranes

  • Yuk, Yeong-Jae;Youm, Kyung-Ho
    • Proceedings of the Membrane Society of Korea Conference
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    • 2004.05a
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    • pp.59-62
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    • 2004
  • Affinity membranes have emerged principally to overcome the problems of limited specificity experienced with membranes that operate purely on a sieving mechanism and as an alternative to the traditional affinity resins. It is a logical expectation that affinity membranes might combine the outstanding selectivity of affinity resins with the high productivity associated with filtration membranes.(omitted)

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Purification of a Mosquitocidal Toxic Protein from B. thuringiensis strain H9B by Immuno-Affinity Chromatography (Immuno-Affinity Chromatography에 의한 B. thuringiensis H9B 균주의 모기살충성 내독소 단백질의 정제)

  • 김광현;배수장;이광배
    • Journal of environmental and Sanitary engineering
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    • v.12 no.2
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    • pp.59-64
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    • 1997
  • For purification of a 70kDa toxic protein of mosquitocidal delta-endotoxin from B. thuringiensis strain H9B, immuno-affinity chromatography was performed. After separation of 70kDa toxic proteins from the delta-endotoxin of the strain H9B on SDS-PAGE, the 70kDa toxic protein was subcutaneously injected into rabbit for making a polyclonal antibody. A anti-70kDa toxic protein was purified by a column chromatography packed with protein A-sepharose 4B gels. The 70kDa toxic protein from delta-endotoxin of the strain H9B was also purified by an immuno-affinity chromatography packed with CNBr-activated sepharose 4B gels conjugated anti-70kDa toxic protein after elution with 1/10M citric acid-1/5M Na$_{2}$HPO$_{4}$ buffer(pH3.2) containing 0.5M NaCl. The 70kDa toxic protein was purified through only one step-separation system, was demonstrated by SDS-PAGE and immunoblot.

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Affinity Separations Using Microfabricated Microfluidic Devices: In Situ Photopolymerization and Use in Protein Separations

  • Chen Li;Lee, Wen-Chien;Lee, Kelvin H.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.4
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    • pp.240-245
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    • 2003
  • The use of microfabricated microfluidic devices offers significant advantages over current technologies including fast analysis time and small reagent requirements. In the context of proteomic research, the possibility of using affinity-based separations for prefractionation of samples using microfluidic devices has significant potential. We demonstrate the use of microscale devices to achieve affinity separations of proteins using a device fabricated from borosilicate glass wafers. Photolithography and wet etching are used to pattern individual glass wafers and the wafers are fusion bonded at 650$^{\circ}C$ to obtain enclosed channels. A polymer has been successfully polymerized in situ and used either as a frit for packing beads or, when derivatized with Cibacron Blue 3GA, as a separation matrix. Both of these technologies are based on in situ UV photopolymerization of glycidyl methacrylate (GMA) and trimethylolpropane trimethacrylate (TRIM) in channels.

Performance Test of Silicone Rubber Membrane by Gas Permeation Method (기체투과에 의한 Silicone Rubber Membrane의 기능성 시험)

  • Lee, Seung-Bum;Hong, In-Kwan
    • Elastomers and Composites
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    • v.33 no.1
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    • pp.37-43
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    • 1998
  • The permeation of gas through polymer membrane at temperatures above its glass transition, generally occurs by a solution-diffusion mechanism. This mechanism is performed by the affinity difference between polymeric materials and gas molecules, and various technologies, such as copolymerization, impregnation and so on, have been researched to improve the affinity of polymeric material for the gases. In this study, permeability and selectivity for some gases were obtained from steady-state rates of gas permeation through silicone rubber membrane which is prepared by supercritical fluid extraction method. The permeability was measured by the volumetric method proposed by Barrer. Permeability was increased generally with temperature and permeation pressure. Silicone rubber membrane shows a higher permeability to $CO_2$ than to $O_2$, $N_2$. This results probably reflect the relatively high solubility of CO_2 in silicone rubber membrane, which is due to the affinity of $CO_2$ molecules. Since separation powers of $CO_2/N_2$, $CO_2/O_2$ were more than 200, and 100, respectively, it is able to separate $CO_2$ from the air, and the optimum temperature and pres-sure was 328.15 K, 60 cmHg respectively. In future, it is possible that the silicone rubber membrane can be used for separation or concentration of $CO_2$ through experiment for mixed gas separation.

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Development of the Purification Method of Ovotransferrin in Egg White (난백 내 Ovotransferrin의 분리방법에 관한 연구)

  • Jang, A.;Jo, Y.J.;Lee, M.;Kim, J.C.
    • Journal of Animal Science and Technology
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    • v.47 no.6
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    • pp.1025-1032
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    • 2005
  • This study was carried out to separate ovotransferrin in chicken egg white by gel chromatography and heparin affinity chromatography. In gel filtration which was performed with 50mM Phosphate buffer (pH 7.2, 0.15M salt) at a flow rate of 2.0 ml/min, ovotransferrin and ovalbumin were eluted together in fraction number 11-16. In order to separate pure ovotransferrin, fraction No. 12-14 of them which have high concentration of ovotransferrin were concentrated and rechromatographed. However, the ovotransferrin did not separated clearly. In heparin affinity chromatography, the separation was performed with 50mM ethylaminetetraacetic acid (EDTA, pH7.2) and 50mM Phosphate buffer (pH 7.2, 0.15M salt contained) on ferrous and ferric ion saturated column at as same flow rate as gel filtration system's. Ovotransferrin and albumin were eluted together at 10-15min (fraction No.3) and 15-20min (fraction No.4), respectively. However, purified ovotransferrin was eluted at 156-165min and 165-175min (tube No.32-33) with 50 mM phosphate buffer (pH 7.2, 0.15M salt free), respectively. Heparin affinity chromatography with ferric ion saturated column was resulted in the best separation of ovotransferrin rather than separation by gel chromatography and ferrous ion saturated heparin affinity chromatography.