• Korean Journal of Microbiology
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• v.41 no.3
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• pp.216-224
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• 2005

Chromatography has now been used successfully to provide the requisite purity for human plasma-derived biop-harmaceuticals such as coagulation factors and immunoglobulins. Recently, increasing attention has been focused on establishing efficient cleaning procedures to prevent potential contamination by microorganisms as well as carry-over contamination from batch to batch. The purpose of present study was to develop a cleaning validation system for the assurance of virus removal and/or inactivation during chromatography process. In order to establish an assay system for the validation of virus clearance during chromatography cleaning process, a quantitative real-time PCR method for porcine parvovirus(PPV) was developed, since PPV, a model virus for human parvovirus B19, has a high resistance to a range of physico-chemical treatment. Specific primers for amplification of PPV DNA was selected, and PPV DNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be 1.5 $TCID_{50}/ml$. The established real-time PCR assay was successfully applied to the validation of PPV removal and cleaning during SP-Sepharose cation chromatography for thrombin purification and Q-Sepharose anion chromatography for factor VIII purification. The comparative results obtained by real-time PCR assay and infectivity titrations suggested that the real-time PCR assay could be a useful method for chromatography cleaning validation and that it could have an additive effect on the interpretation and evaluation of virus clearance during the virus removal process.

• KSBB Journal
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• v.9 no.4
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• pp.385-394
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• 1994

Large scale purification to get antifungal antibiotic KRF-001 of 90% purity, was investigated using preparative HPLC. Crude KRF-001 was purified by XAD-7 adsorption chromatography, acid precipitation and microfiltration. Microfiltration was the most effective isolation method of crude KRF-001. The purification methods using C18 chromatography was convenient compared with the conventional methods. Delta PAK C18 column and Bonda PAK C18 column were adapted large scale purification of KRF-001. Gradient system of prep HPLC using Delta PAK C18 column was more effective. With these conditions, final recovery of KRF-001 yielded 77%.

• Journal of the Korean Chemical Society
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• v.48 no.2
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• pp.203-206
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• 2004

• Proceedings of the Korean Society of Applied Pharmacology
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• 1992.05a
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• pp.22-22
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• 1992

국내의 토양에서 Streptomyces spp.를 분리하였고 이 중 항균활성이 우수한 균주들을 선별하였다. 이들 균주들을 적절한 조건에서 배양하여 얻은 배양액을 이온 교한 수지와 흡착 크로마토그래피를 행하여 항생물질들을 분리하였다. 이들 항생물질들은 Bacillus subtilis, Pyricularia oryzae, Alternaria mail, Candida albicans, Mycobacterium smegmatis, Pseudomonas aeruginosa , 및 Escherichia coli에 강한 항균활성을 나타내었다. 각 항생물질들을 산으로 가수분해하고 흡착관 크로마토그래피로 가수분해 생성물을 분리하였다. 분리한 항생물질과 가수분해 생성물들의 $^1$IH-NMR, $13^{C}$ -NMR, FAB-Mass 스펙트럼을 분석하여 구조를 연구하였다.

• 대한방사선방어학회:학술대회논문집
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• 2010.04a
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• pp.84-85
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• 2010

• Journal of the Korean Chemical Society
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• v.52 no.4
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• pp.423-433
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• 2008

• KSBB Journal
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• v.14 no.3
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• pp.279-283
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• 1999

We have compared the breakthrough behavior of lysozyme contained in fresh han egg white on various cation exchagers, and the adsorbent, known by the trade name Cellufine C-200 (Amicon), has shown the best performance. The effects of ion strength, pH, and linear flow rate on the breakthrough behavior were examined using the Cellufine C-200 adsorbent. The optimal conductivity, pH and linear flow rate were determined from the breakthrough behavior and found to be 2.75 mS/cm, 7.0, and 0.635 cm/min, respectively.

• Applied Chemistry for Engineering
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• v.7 no.5
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• pp.985-991
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• 1996

In chromatographic separation, HETP(height equivalent to a theoretical plate) is a useful quanititive parameter and it is wildely designated as column efficiency. The effects of operating conditions (sample concentration, injection volume, flow rate and mobile phase composition) on HETP were investigated in perparative liquid chromatography (PLC). Water and organic modifier of methanol were used as mobile phase. The sample of thymidine was injected into preparative C18 columns. The system was run by a isocratic mode in 1.5~5.5ml/min. The larger amounts of sample and higher flow rates of mobile phase increased HETP, which means that column efficiencies were worse. As the weight of sample injected into a chromatographic system could be prepared with different concentrations and injection volumes, for the same amount of sample, HETP was approximately increased two times with the ten-fold injection volume. HETP was mainly affected by the resistance of stationary and mobile phase mass transfer in the intraparticle section of packings at higher velocities.

• KSBB Journal
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• v.22 no.5
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• pp.366-369
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• 2007

In this study, the feasibility of reusing the eluent was confirmed by monitoring its specific gravity during the chromatographic purification of paclitaxel from plant cell cultures. The specific gravity of the eluent (methanol/water = 70/30, v/v) was measured prior to its elution through the hydrophobic resin column. The measurement showed a specific gravity of 0.853. The discharged eluent from the column outlet was first evaporated under vacuum pressure. The evaporated eluent was collected and condensed into a liquid eluent again, followed by the HPLC analysis in order to check the presence of any trace of impurity. Even if the specific gravity of the liquid eluent is varied from 0.853 as a result of the evaporation and condensation, the eluent can still be reused after it specific gravity is adjusted by the addition of methanol or water. The reuse of the eluent resulted in the paclitaxel yield of 86% with a purity of 95% which were closely similar to those of before the eluent reuse. These results indicate that the strategy of reusing the eluent on the basis of the specific gravity analysis was successfully implemented in this study.

• Membrane Journal
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• v.8 no.1
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• pp.50-58
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• 1998

Protein affinity membranes were prepared via coating of chitosan gel on the porous flat and hollow-fiber polysulfone membranes, followed by the immobilization of the reactive dye (Cibacron Blue 3GA) to the chitosan gel. Maximum protein binding capacity of these affinity membranes was about 70 $\mu{g/cm}^2$. Using the affinity flat membrane module, the elution chromatography of human serum albumin (HSA) was performed to determine the optimum condition of eluent buffer. The optimum condition of eluent was the universal buffer solution of 0.06 M concentration containing 1 M KCl at pH 10. For the frontal chromatography of HSA using the flat module, the dynamic protein binding capacity was rapidly decreased from the equilibrium values with increasing flow rate and HSA concentration of the loading solution. However, in the case of hollow-fiber module, the dynamic binding capacity was maintained an equilibrium value without depending on the operating conditions. These results showed that the hollow-fiber module was more effective than the flat module as an affinity chromatography column.