• Title/Summary/Keyword: continuous chromatography

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Development of Chromatographic Downstream Processing for The Purification of Monoclonal Antibody from Ascites Fluid: Part 1. Tandem Use of Hydroxylapatite Chromatography and Gel Permeation Chromatography (복수로부터의 단세포군 항체의 정제를 위한 크로마토그래프 분리 정제 시스템의 개발 - 1. 히드록실 아파타이트 크로마토그라프와 겔 여과 크로마토그라프로 이루어진 2단계 연속공정 사용 -)

  • Ahn, I.S.;Park, C.Y.
    • Microbiology and Biotechnology Letters
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    • v.17 no.1
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    • pp.19-23
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    • 1989
  • A sequential system composed of hydroxylapatite chromatography and gel permeation chromatography was developed to purify the IgM type monoclonal antibody against the colon cancer cell SC-1 from the ascitic fluid of mice injected with the murine hybridoma CH07E02. In the hydroxylapatite chromatographic step the band dilution could be reduced by controlling the gradient and flow rate of the eluent, the sodium phospate buffer, the optimum values for these variables being 5.82$\times$10$^{-3}$M/cm and 0.2$m\ell/\textrm{cm}^2$/min, respectively. A degree of purity better than 99.99% as judged from silverstaining of the SDS-PAGE bands, was obtained by adding the gel permeation chromatographic step in tandem.

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Microprocessor Based Laser Induced Fluorometry I. Development of System and Application to Liquid Chromatography

  • Kim, Ha-Suck;Park, Chan-Seung;Hwang, Eui-Jin;Chol, Q.-Won
    • Bulletin of the Korean Chemical Society
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    • v.5 no.6
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    • pp.253-259
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    • 1984
  • An analytical applicability of the fluorescence detection with an optical multichannel analyzer to organic dyes was studied in this work. Continuous acquisition of spectra was possible with the use of a microcomputer. The maximum acquisition rate of a spetrum with 70-point average was about 3 seconds. Floppy discs were used to store data for later use in processing. Laser induced fluorescence detector in High Performance Liquid Chromatography was chosen for an application. Fluorescein below $10^{-15}g$ was detected satisfactorily using this system. With the help of microcomputer, three dimensional chromatograms of time-wavelength-intensity were obtained.

Improved Large-Scale Refolding Techniques for Inclusion Body Proteins (내포체 단백질의 개선된 대규모 재접힘 기술)

  • 김인호;정봉현
    • KSBB Journal
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    • v.16 no.1
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    • pp.11-14
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    • 2001
  • Techniques for protein refolding from inclusion body are discussed in view of its engineering application to large scale protein purification. Among the techniques, dilution and dialysis are mainly utilized due to simple operation. Membrane reactor, gel filtration chromatography, and continuous tank operation are emerging tools for their process-scale possibility in refolding. Reaction engineering approaches could be used to analyze the kinetic behaviour in the process scale refolding reactor. The kinetic analysis is helpful in the optimization of refolding yield in the refolding reactor.

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Procedures for Analyzing Ethylene by Gas Chromatograph (Gas Chromatograph를 이용한 에틸렌 분석 기술)

  • 이승구
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.34 no.s01
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    • pp.33-39
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    • 1989
  • Ethylene gas classified as one of five major plant hormones plays an important role in various plant metabolism. The precise analysis of ethylene production of plants or plant parts is a valuable research procedure because knowledge of ethylene production facilitates measures of the physiological activity within the tissue. This paper describes procedures for analyzing ethylene from plant tissues by gas chromatography and discusses problems associated with extracting gas samples either by introducing a vacuum to plant samples or by using a hypodermic syringe. Introduced are a continuous flow system for efficient analysis and an automated system for sampling, analyzing, calculating and recording ethylene production data.

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Purification of Isocitrate lyase Produced from Microbacterium laevaniformans (Microbacterium laevaniformans가 생성하는 Isocitrate lyase의 정제)

  • 서승교;김정호
    • Journal of Environmental Science International
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    • v.7 no.6
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    • pp.853-857
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    • 1998
  • Purification of the isocitrate lyase extracted from Microbacterium laevaniformans was investigated. The isocitrate lyase was purified 43.6 folds by the following continuous treatment with ammonium sulfate fraction, DEAE-cellulose, DEAE-sephacel and Sephadex G-200 chromatography. The purified isocitrate lyase was showed to be a single protein band by polyacrylamide gel electrophoresis. The molecular weight of the purified isocitrate lyase was estimated 54,000 Da by the SDS-polyacrylamide gel electrophoresis. The Km and Vmax values for isocitrate were estimated to be 0.83mM and 0.33units/ml, respectively. Activity of isocitrate lyase was inhibited by cystein-HCl and glutathione.

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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).

Comparative Simulation of 3-zone SMB (Simulated Moving Bed) and 4-zone SMB for IgY (Immunoglobulin Yolk) Purification (IgY (Immunoglobulin Yolk) 분리를 위한 3-영역 SMB (Simulated Moving Bed)와 4-영역 SMB 비교전산모사)

  • Yun, Sang-Hee;Kim, In-Ho
    • Korean Chemical Engineering Research
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    • v.50 no.5
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    • pp.866-873
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    • 2012
  • IgY (Immunoglobulin Yolk) is a specific antibody in egg yolk, and it protects human body from virus and antigen. There are a lot of egg yolk components such as lipoprotein and protein. To separate IgY, HPLC (High Performance Liquid Chromatography) and precipitation were used in a batch mode and SMB (Simulated Moving Bed) was adopted for continuous purification of yolk proteins. IgY and other proteins in yolk were separated by using three-zone and four-zone SMB chromatography. Before performing SMB experiments, batch chromatography simulation parameters and adsorption isotherms were obtained. The parameters of batch chromatography were used to simulate SMB using Aspen chromatography. To compare three-zone and four-zone SMB chromatography, simulations in $m_2-m_3$ plane on the triangle theory were carried out. In terms of concentration and purity of both IgY and other lipoproteins, 3-zone SMB process is considered as ideal at the vertex of triangle ($m_2$, $m_3$=0.1, 1.1). 4-zone SMB yields the highest IgY purity at the coordinate ($m_2$, $m_3$=0.06, 0.5), which is the pure raffinate region. In 3-zone SMB without recycle, other lipoproteins in extract are largely affected in purity by small shift from the vertex of triangle ($m_2$, $m_3$=0.1, 1.1).

Identification of Factors Regulating Escherichia coli 2,3-Butanediol Production by Continuous Culture and Metabolic Flux Analysis

  • Lu, Mingshou;Lee, Soo-Jin;Kim, Bo-Rim;Park, Chang-Hun;Oh, Min-Kyu;Park, Kyung-Moon;Lee, Sang-Yup;Lee, Jin-Won
    • Journal of Microbiology and Biotechnology
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    • v.22 no.5
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    • pp.659-667
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    • 2012
  • 2,3-Butanediol (2,3-BDO) is an organic compound with a wide range of industrial applications. Although Escherichia coli is often used for the production of organic compounds, the wild-type E. coli does not contain two essential genes in the 2,3-BDO biosynthesis pathway, and cannot ferment 2,3-BDO. Therefore, a 2,3-BDO biosynthesis mutant strain of Escherichia coli was constructed and cultured. To determine the optimum culture factors for 2,3-BDO production, experiments were conducted under different culture environments ranging from strongly acidic to neutral pH. The extracellular metabolite profiles were obtained using high-performance liquid chromatography (HPLC), and the intracellular metabolite profiles were analyzed by ultra-performance liquid chromatography and quadruple time-of-flight mass spectrometry (UPLC/Q-TOF-MS). Metabolic flux analysis (MFA) was used to integrate these profiles. The metabolite profiles showed that 2,3-BDO production favors an acidic environment (pH 5), whereas cell mass favors a neutral environment. Furthermore, when the pH of the culture fell below 5, both the cell growth and 2,3-BDO production were inhibited.

The effect of supercritical water treatment on the chemical variations of lignin (목질바이오매스의 초임계수 처리에 의한 리그닌의 화학적 변환)

  • Lee, Soo-Min;Lee, Oh-Kyu;Choi, Seok-Hwan;Choi, Joon-Weon;Choi, Don-Ha
    • 한국신재생에너지학회:학술대회논문집
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    • 2008.05a
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    • pp.206-209
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    • 2008
  • The modified supercritical water treatment method is adopted for hydrolysis of wood powder, Populus alba$\times$glandulosa. This modified method is containing 0.05% HCl or $HNO_3$ as acid catalyst. The supercritical water treatment(SCW) was performed for 1 min. with $350^{\circ}C$, $380^{\circ}C$, $400^{\circ}C$ and $425^{\circ}C$, respectively, under 230 $\pm$ 10 atm using continuous flow system. When acid was added to powder prepared for SCW treatment, the yields of monomeric sugars were significantly increased. The lignin remained after supercritical treatment was applied to gel permeation chromatography(GPC) for molecular weight distribution analysis. Compared to the lignin produced from SCW treatment without acid catalyst, the average molecular weight of lignin compounds treated with acid was clearly decreased. Particularly, Mn/Mw ratio is decreased. This result shows supercritical water treatment of wood powder can change the molecular weight of lignin to small size. However, it is necessary to be further studied for exactly characterizing the lignin produced from supercritical water treatment.

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The Development of VOC Measurement System Uging PCA & ANN (PCA와 ANN을 이용한 VOC 측정기기 개발)

  • Lee Jang-Hoon;Kwon Hyuk-Ku;Park Seung Ho;Kim Dong-Jin;Hong Chol-Ho
    • Environmental Analysis Health and Toxicology
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    • v.19 no.2
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    • pp.161-167
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    • 2004
  • Air quality monitoring is a primary activity for industrial and social environment. The government identifies the pollutants that each industry must monitor. Especially, the VOCs (Volatile Organic Compounds), which are very harmful to human body and environment atmosphere, should be controlled under the government policy. However, the VOCs, which have not been confirmed in emission sources are very difficult to monitor. It is needed to develop the monitoring system that allow the continuous and in situ measurement of VOCs mixture in different environmental matrices. Gas chromatography and mass spectrometry are the most prevalent current techniques among those available for the analysis of VOCs. But, they need a large size analytical instrument, which costs a great deal for purchase and operation. In addition, it has some limitations for realtime environmental monitoring such as location problems and slow processing time. Recently, several companies have commercialized a portable VOCs measurement systems, which cannot classify various kinds of VOCs but total quantities. We have developed a VOCs measurement system, which recognizes various kinds and quantities of VOCs, such as benzene, toluene, and xylene (BTX). Also, it can be used as a stand- alone type and/or fixed type in the vehicle with rack for real -time environmental monitoring.