• 제목/요약/키워드: size exclusion

검색결과 245건 처리시간 0.032초

A Preparative Purification Process for Recombinant Hepatitis B Core Antigen Using Online Capture by Expanded Bed Adsorption Followed by Size-Exclusion Chromatography

  • Ho, Chin Woi;Tan, Wen Siang;Chong, Fui Chin;Ling, Tau Chuan;Tey, Beng Ti
    • Journal of Microbiology and Biotechnology
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    • 제19권4호
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    • pp.416-423
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    • 2009
  • Hepatitis B core antigen(HBcAg) is an important serological marker used in the diagnosis of hepatitis B virus(HBV) infections. In the current study, a fast and efficient preparative purification protocol for truncated HBcAg from Escherichia coli disruptate was developed. The recombinant HBcAg was first captured by anion exchange expanded bed adsorption chromatography integrated with a cell disruption process. This online capture process has shortened the process time and eliminated the "hold-up" period that may be detrimental to the quality of target protein. The eluted product from the expanded bed adsorption chromatography was subsequently purified using size-exclusion chromatography. The results showed that this novel purification protocol achieved a recovery yield of 45.1% with a product purity of 88.2%, which corresponds to a purification factor of 4.5. The recovered HBcAg is still biologically active as shown by ELISA test.

Comparison of Size-Exclusion Chromatography and Flow Field-Flow Fractionation for Separation of Whey Proteins

  • Kang, Da-Young;Moon, Jae-Mi;Lee, Seung-Ho
    • Bulletin of the Korean Chemical Society
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    • 제32권4호
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    • pp.1315-1320
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    • 2011
  • Whey protein (WP) is a mixture of proteins, and is of high nutritional values. WP has become an important source of functional ingredients in various health-promoting foods. In this study, size-exclusion chromatography (SEC) and asymmetrical flow field-flow fractionation (AsFlFFF) were used for separation and analysis of whey proteins. It was found that a lab-prepared WP from raw milk is mostly of ${\beta}$-lactoglobulin with small amount of higher molecular weight components, while a commercial whey protein isolate (WPI) powder contains relatively larger amount of components other than ${\beta}$-lactoglobulin, including IgG and protein aggregates. Results suggest that AsFlFFF provides higher resolution for the major whey proteins than SEC in their normal operation conditions. AsFlFFF could differentiate the BSA and Albumin, despite a small difference in their molecular weights, and also was able to separate much smaller amount of aggregates from monomers. It is noted that SEC was able to show the presence of low molecular weight components other than the major whey proteins in the WP samples, which AsFlFFF could not show, probably due to the partial loss of those low molecular weight species through the membrane.

High Temperature Size Exclusion Chromatography

  • Cho Hee-Sook;Park Soo-Jin;Ree Moon-Hor;Chang Tai-Hyun;Jung Jin-Chul;Zin Wang-Cheol
    • Macromolecular Research
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    • 제14권3호
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    • pp.383-386
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    • 2006
  • High temperature size exclusion chromatography (SEC) has been used widely for the characterization of crystalline polymers, for which high temperature operation above the polymer melting temperature is required to dissolve the polymers. However, this high temperature operation has many advantages in SEC separation in addition to merely increasing polymer solubility. At high temperature the eluent viscosity decreases, which in turn decreases the column backpressure and increases the diffusivity of the analytes. Therefore, many reports on the high temperature operation of high performance liquid chromatography (HPLC) have focused on shortening the analysis time and enhancing the resolution. However, the application of high temperature SEC analysis to exploit the merits of high temperature operation is scarce. In this article, therefore, we report on a new apparatus design for high temperature SEC.

Refolding and Purification of Recombinant Human $Interferon-\gamma$ Expressed as Inclusion Bodies in Escherichia coli Using Size Exclusion Chromatography

  • Guan Yi-Xin;Pan Hai-Xue;Gao Yong-Gui;Yao Shan-Jing;Cho Man-Gi
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제10권2호
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    • pp.122-127
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    • 2005
  • A size exclusion chromatography (SEC) process, in the presence of denaturant in the refolding buffer was developed to refold recombinant human $interferon-\gamma$ ($rhIFN-\gamma$) at a high concentration. The $rhlFN-\gamma$ was overexpressed in E. coli resulting in the formation of inactive inclusion bodies (IBs). The IBs were first solubilized in 8 M urea as the denaturant, and then the refolding process performed by decreasing the urea concentration on the SEC column to suppress protein aggregation. The effects of the urea concentration, protein loading mode and column height during the refolding step were investigated. The combination of the buffer-exchange effect of SEC and a moderate urea concentration in the refolding buffer resulted in an efficient route for producing correctly folded $rhIFN-\gamma$, with protein recovery of $67.1\%$ and specific activity up to $1.2\times10^7\;IU/mg$.

공동캡슐화를 이용한 키토산 분해반응에서 alginate 막의 특성 및 크기에 따른 올리고당의 분자량 내외 분포

  • 이기선;최명락;송상호;임현수
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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    • pp.608-611
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    • 2000
  • To separate chitosanoligosaccharides easily by size exclusion, an coencapsulating technology of substrate and enzyme was developed. Chitosan and chitosanase were enveloped in this membrane and the product released to medium by size exclusion. The lower limit of the alginate concentration and the agitation speed were 0.5% and 40 rpm, respectively. Membrane thickness and capsules diameter were $10{\mu}m$ and approx. 3.0mm, 1.5mm, respectively. The molecular weight difference by concentration and cps of alginate were of little significance. And also, the molecular weight of distribution according to enzyme concentration was low concentration of enzyme produced high molecular weight of oligosaccharides. At 1.5mm size of capsule, product diffusion rate to outer part was higher than other capsules. The molecular weight distribution of the released oligosaccharides ranged from 1000 to 6000 Da.

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Studies on the Separation Performances of Chlorophenol Compounds from Water by Thin Film Composite Membranes

  • Yogesh, K.M. Popat;Ganguly, B.;Brahmbhatt, H.;Bhattacharya, A.
    • Macromolecular Research
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    • 제16권7호
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    • pp.590-595
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    • 2008
  • The pressure driven membrane process has been a breakthrough in the removal of pollutants from drinking water. These experiments examined the removal of chlorophenol compounds from water using low pressure membranes. The removal performance of the membranes was based primarily on size exclusion. Apart from size exclusion, the polarity and pKa of the compounds also influences the membrane performance. The molecular size and dipole moments of the respective molecules were calculated using a quantum chemical method. The rejection of pollutants also followed the same trend as salt rejection by the membranes.

Size-exclusion Chromatography를 활용한 가공식품 중 폴리아크릴산나트륨 분석법 확립 (Analytical Method for Sodium Polyacrylate in Processed Food Products by Using Size-exclusion Chromatography)

  • 정은정;최유정;이근영;윤상순;임호수;김미경;김용석
    • 한국식품위생안전성학회지
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    • 제33권6호
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    • pp.466-473
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    • 2018
  • 가공식품에서 폴리아크릴산나트륨에 대한 분석 방법을 size-exclusion chromatography를 사용하여 개발하고 유통 중인 가공식품에 대해 모니터링 하였다. 분석조건에 대하여 GF-7M HQ column과 UV / VIS 검출기를 피크 모양과 선형성을 기준으로 선택하였으며, 유속, column oven 온도 및 이동상은 각각 0.6 mL/min, $45^{\circ}C$ 및 50 mM sodium phosphate buffer (pH 9.0)으로 선정되었다. 시료의 전처리를 위하여 pH 7.0의 50 mM sodium phosphate buffer로 $20^{\circ}C$, 150 rpm에서 3 시간 동안 추출하였다. 확립된 분석법을 검증한 결과 적절한 선택성을 나타내었으며, 검량선은 50~500 mg/L의 범위에서 선정되었고, 이때 검량선의 상관계수($R^2$)는 0.9985로 나타났다. 폴리아크릴산나트륨의 검출한계(LOD)는 10.95 mg/kg으로 정량한계(LOQ)는 33.19 mg/kg의 값을 얻었으며, 정밀도는 intra-day의 경우 3.0~8.3%를, inter day의 경우 1.3~2.6%로 확인되었다. 정확도는 intra-day의 경우 99.6~127.6%를 나타내었으며, interday의 경우 94.3~121.9%로 확인되었다. 유통 중인 가공식품 125품목을 대상으로 폴리아크릴산나트륨의 함유량을 분석한 결과 40품목에서 검출되었으며, 검출량은 식품첨가물공전의 규격인 0.2% 미만으로 검출되었다. 본 연구결과는 size-exclusion chromatography에 의한 분석방법이 가공식품에서 폴리아크릴산나트륨을 분석하는데 적용할 수 있는 적합한 방법이라는 것을 나타내었다.

사회적 배척과 소속 욕구가 사회적 사건의 정서 예측에 미치는 영향 (The impacts of social exclusion and the need to belong on the affective forecasting of social events)

  • 김애리;손영우;임혜빈
    • 감성과학
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    • 제17권3호
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    • pp.83-94
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    • 2014
  • 본 연구는 개인의 사회적 배척 경험과 소속 욕구 수준이 사회적 사건에 대한 정서 예측 및 정서 예측 오류의 크기에 미치는 영향을 검증하기 위하여 실시되었다. 참가자들은 예비 연구를 통해 구성된 8가지 종류의 사회적 사건과 비사회적 사건에 대해 각 사건이 실제로 벌어진다면 얼마나 기쁠지 혹은 괴로울지에 대해 응답하였다. 연구 1에서는 사회적 배척 경험이 많을수록 또는 소속 욕구가 높을수록 비사회적 사건에 비해 사회적 사건이 가지고 올 괴로움을 더욱 부정적으로 예측하는 것으로 나타났다. 그러나 연구 2에 의하면, 사회적 배척 경험이 많거나 소속 욕구가 높다고 해서 실제로 사회적 부정적 사건을 경험했을 때 더 괴로운 것은 아니라는 것이 밝혀졌다. 본 연구의 결과는 사회적 배척 경험이 많거나 소속 욕구가 높은 개인의 사회적 사건에 대한 민감한 정서 예측이 그들이 사회적 사건으로부터 경험하는 실질적 괴로움이 더 크기 때문이 아니라 사회적 사건으로 인해 겪게 될 미래 정서를 과대 추정하기 때문이라는 것을 시사한다. 본 연구 결과의 함의와 제한점, 추후 연구에 대한 논의가 제시되었다.

High Temperature Size Exclusion Chromatography for High Throughput Analysis

  • Chang, Tai-Hyun;Park, Soo-Jin;Cho, Hee-Sook;Kim, Young-Tak;Ihm, Kyu-Hyun
    • 한국고분자학회:학술대회논문집
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    • 한국고분자학회 2006년도 IUPAC International Symposium on Advanced Polymers for Emerging Technologies
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    • pp.202-202
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    • 2006
  • With a modern size exclusion chromatography (SEC) column, molecular weight analysis of a polymer sample takes about 10 min. However, it is desirable to reduce the analysis time further, in particular, for high throughput measurements required in combinatorial analyses or 2D-HPLC analyses. We implemented the high temperature SEC for the purpose. By inserting a narrow bore tubing between the column and the detector, a sufficient backpressure can be maintained to prevent the mobile phase from boiling and the effluent is cooled down enough when it reaches the detector. Therefore, a normal SEC detector can be used without any modification. The SEC resolution is greatly improved at the elevated temperature at high flow rate which allows high speed operation.

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Mid-term (2009-2019) demographic dynamics of young beech forest in Albongbunji Basin, Ulleungdo, South Korea

  • Cho, Yong-Chan;Sim, Hyung Seok;Jung, Songhie;Kim, Han-Gyeoul;Kim, Jun-Soo;Bae, Kwan-Ho
    • Journal of Ecology and Environment
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    • 제44권4호
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    • pp.241-255
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    • 2020
  • Background: The stem exclusion stage is a stage of forest development that is important for understanding the subsequent understory reinitiation stage and maturation stage during which horizontal heterogeneity is formed. Over the past 11 years (2009-2019), we observed a deciduous broad-leaved forest in the Albongbunji Basin in Ulleungdo, South Korea in its stem exclusion stage, where Fagus engleriana (Engler's beech) is the dominant species, thereby analyzing the changes in the structure (density and size distributions), function (biomass and species richness), and demographics. Results: The mean stem density data presented a bell-shaped curve with initially increasing, peaking, and subsequently decreasing trends in stem density over time, and the mean biomass data showed a sigmoidal pattern indicating that the rate of biomass accumulation slowed over time. Changes in the density and biomass of Fagus engleriana showed a similar trend to the changes in density and biomass at the community level, which is indicative of the strong influence of this species on the changing patterns of forest structure and function. Around 2015, a shift between recruitment and mortality rates was observed. Deterministic processes were the predominant cause of tree mortality in our study; however, soil deposition that began in 2017 in some of the quadrats resulted in an increase in the contribution of stochastic processes (15% in 2019) to tree mortality. The development of horizontal heterogeneity was observed in forest gaps. Conclusions: Our observations showed a dramatic shift between the recruitment and mortality rates in the stem exclusion stage, and that disturbance increases the uncertainty in forest development increases. The minor changes in species composition are likely linked to regional species pool and the limited role of the life-history strategy of species such as shade tolerance and habitat affinity. Our midterm records of ecological succession exhibited detailed demographic dynamics and contributed to the improvement of an ecological perspective in the stem exclusion stage.