• Korean Chemical Engineering Research
• /
• v.43 no.2
• /
• pp.230-235
• /
• 2005

Reversed-phase high-performance liquid chromatography (RP-HPLC) was used to determine the equilibrium isotherm of single and multi-components of dUrd(2'-deoxyuridine), dGuo(2'-deoxyguanosine), and dAdo(2'-deoxyadenosine) of 2'-deoxyribonucleosides by dynamic method. The composition of mobile phase was 90/10 vol.% (water/MeOH). With an increase in the injection volumes, the retention times were shorter and the peak shapes were triangle-shaped, so Langmuir-type isotherm was assumed. The Langmuir adsorption parameters were estimated by PIM (pulsed-input method), and the competitive Langmuir adsorption isotherm was further utilized. For the sample of the dUrd and dGuo whose retention times were relatively short, the agreement of between the calculated value and experimental data was fairly good in both single and multi-components, but for the dAdo, the last eluting component, some deviations were caused by non-linear and non-ideal properties.

• Analytical Science and Technology
• /
• v.20 no.5
• /
• pp.442-447
• /
• 2007

The properties of garlic (Allium sativum L.) are attributed to organosulfur compounds. In this paper, an analytical technique with a rapid and simple sample preparation procedures for determination of diallyl disulfide (DADS) in garlic was reported. The DADS was simply extracted with various solvents (methanol, benzene or tetrahydrofuran) from garlic and prepared for HPLC analysis. From the results, the methanol was select as an optimal extraction solvent. The mobile phase was composed from methanol and water, and the gradient elution mode was applied. 0.61 mg of DADS per g garlic powder can be extracted with methanol. This work offers some advantages over the currently accepted techniques and would be useful for chemical and biological studies of garlic and its products.

• Microbiology and Biotechnology Letters
• /
• v.35 no.3
• /
• pp.226-230
• /
• 2007

In order to achieve high-level expression of interferon-${\alpha}1$ (IFN-${\alpha}1$) during fed-batch fermentation of recombinant E. coli, effects of oxygen supply and induction temperature on the expression of recombinant proteins were evaluated. Supplementation of oxygen and its transfer into cells is one of the most important parameters involved in the design and operation of mixing-sparging equipment for bioreactors. Generally, higher oxygen supply stimulates cell growth of aerobic microorganism and consequently the amount of products is increased. In this study, the optimum aeration strategy for the higher production of IFN-${\alpha}1$ during fed-batch fermentation of recombinant E. coli was surveyed. The growth of the cells was also monitored with four different concentrations of dissolved oxygen (DO; limiting, 20%, 35%, 50%) conditions. The DO was controlled by varying aeration rates of air and pure oxygen. Oxygen uptake rate (OUR) and specific oxygen uptake rate (SOUR) were evaluated and compared for the enhanced growth and induction of the cells and IFN-${\alpha}1$, respectively. We confirmed that increased DO by additional oxygen supply, up to 35%, can improve the production of IFN-${\alpha}1$ during the fermentation.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.9 no.6
• /
• pp.495-499
• /
• 2004

The analyses of peak shapes in chromatography are useful in operating chroma­tographic system. The asymmetry and sharpness of a chromatographic peak are estimated by the reversed-phase adsorption of two standard peptides (angiotensin II bradykinin) on $C_{18}$ In this work, the average particle diameters of $C_{18}$ were 5 and 15 $\mu$m, while the pore sizes were 100 and 300 A. The composition of the mobile phase was $50/50\;vol.\;{\%}$ of a binary mixture of acetonitrile and water with $0.1\%$ TFA, and the particles were packed in a stainless column ($4.6{\times}150$ mm). The third and the fourth central movement were calculated from the chromatographic elution curves by moment analysis. The peak asymmetry was determined by two theoretical calculations: the asymmetry factor by elution peak analysis and skewness with moment analysis. The sharpness was estimated by the fourth central moment. In this work, the most acceptable skewness was calculated when the pore size was 300 A. The large excess was observed on small pore size.

• Journal of Microbiology
• /
• v.42 no.3
• /
• pp.243-247
• /
• 2004

Sphingomonas chlorophenolica ATCC39723 was successfully labeled with the gfp (green fluorescent protein) gene inserted into the pcpB gene by homologous recombination. As the gfp recombinant was easily distinguished from other indigenous organisms, the population of gfp recombinant was monitored after being released into the soil microcosms. Their population density dropped from 10$\^$8/ to 10$\^$6/ (cfu/$m\ell$) in the non-sterilized soil microcosms during the first 6 days. Moreover, the gfp recombinant was not detected even at lower dilution rates after a certain time period. The recombinant, however, survived for at least 28 days in the sterilized soil microcosms. Although the gfp recombinant did not degrade pentachlorophenol (PCP), this experiment showed the possibility of using gfp as a monitoring reporter system for S. chlorophenolica ATCC39723 and potentially other species of Sphingomonas.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.6 no.1
• /
• pp.37-41
• /
• 2001

HPLC separation of ionic samples tends to be more complicated and difficult to understand than that of non-ionic compounds. On the other hand, band spacing is much more easily manipulated for ionic than for neutral samples. Ion-suppression RP-HPLC method was used with organic modifier and aqueous buffer solution. In this work, five mononucleotides of cytidine-5-monophosphate (5-CMP) disodium salt, uridine-5-monophosphate disodium salt (5-UMP), guanosine-5-monophosphate disodium salt (5-GMP), inosine-5-monophosphate disodium salt (5-IMP), and adenosine-5-monophosphate disodium salt (5-AMP) were examined. Acetic acid and sodium phosphate were used as buffers, and methanol as an organic modifier. A new relationship between the retention factor and the buffer concentration at a fixed modifier content (5% of methanol) could be expressed by following: K = (k(sub)-1 + k(sub)0 (k(sub)B/k(sub)S)/(1 + (k(sub)B/k(sub)S) C(sub)B(sup)a), where C(sub)B was the concentration of buffer. Using this relationship, the calculated values closely matched the experimental data. The derived relationship showed that as the buffer concentration increased, the retention factor approached a certain value, and this was buffer dependent.

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.6 no.1
• /
• pp.72-74
• /
• 2001

Transgenic Nicotiana tabacum cells were cultivated for the production of murine granulocyte macrophage-colony stimulating factor (mGM-CSF) in both a stirred tank bioreactor and an airlift bioreactor with draft tube. Cell growth and mGM-CSF production in the airlift bioreactor were found to be better than those achieved in the stirred tank bioreactor. In the airlift bioreactor, 9.0g/L of cells and 2.2ng/mL of mGM-CSF were obtained (11.0g/L and 2.4ng/mL, respectively in shake flasks). Although the lag period was prolonged and mGM-CSF production was lowered by 33% in the stirred thank bioreactor as compared to the control culture, the maximum cell density was increased up to 12.0g/L due to better mixing by agitation at the higher cell density.

• Bulletin of the Korean Chemical Society
• /
• v.28 no.4
• /
• pp.601-606
• /
• 2007

The chromatographic behaviors of nucleotides (inosine 5'-monophosphate, uridine 5'-monophosphate, guanosine 5'-monophosphate, and thymine monophosphate disodium salts) on a C18 column were studied with different types of ionic liquids (ILs) as additives for the mobile phase in reversed-phase liquid chromatography (RPLC). Three ILs, 1-butyl-3-methylimidazolium tetrafluoroborate ([BMIm][BF4]), 1-ethyl-3-methylimidazolium tetrafluoroborate ([EMIm][BF4]), and 1-ethyl-3-methylimidazolium methylsulfate ([EMIm][MS]), were used. Eluents were composed of water and methanol (90/10%, vol) with the addition of 0.5-13.0 mM of ILs. The effects of the concentration of ILs on retention and separation were investigated and discussed. The results showed that the addition of ILs affects the retention and resolution of the tested compounds. Use of 13.0 mM of [BMIm][BF4] as the eluent modifier resulted in a baseline separation of nucleotides without requiring gradient elution. This study demonstrates that ILs can be potentially applied as a mobile phase modifier in RPLC.

• Bulletin of the Korean Chemical Society
• /
• v.28 no.2
• /
• pp.276-280
• /
• 2007

In this work, caffeine and some catechin compounds + C, EC, EGC, and EGCG were extracted from green tea by using molecular imprinted polymers (MIP) as sorbent materials in a solid-phase extraction (SPE) process known as MISPE (molecular imprinted solid-phase extraction). For synthesis of MIP, caffeine was employed as the template, MAA as the monomer, EGDMA as the crosslinker, and AIBN as the initiator. A solution of caffeine (0.2 mg/mL in methanol) was utilized in the solid extraction cartridges following loading, washing, and elution procedures with acetonitrile, methanol, and methanol-acetic acid (90/10, %v/v) as the solvents, respectively. This solid-phase extraction protocol was applied for the extraction of caffeine and some catechin compounds from green tea. A comparison was made between the results obtained with the MIP cartridges and a traditional C18 reversed-phase cartridge. It was thereupon found that the recovery of caffeine by the MIPbased sorbent used in this work was almost two and four times greater than that by a commercially available C18 material. A quantitative analysis was conducted by high performance liquid chromatography (HPLC) using a C18 column (5 μm, 250 × 4.6 mm) with methanol/water (40/60, %v/v) as the mobile phase at a flow rate of 0.5 mL/min.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.3
• /
• pp.349-357
• /
• 2021

Monoclonal antibodies are widely used as diagnostic reagents and for therapeutic purposes, and their demand is increasing extensively. To produce these proteins in sufficient quantities for commercial use, it is necessary to raise the output by scaling up the production processes. This review describes recent trends in high-density cell culture systems established for monoclonal antibody production that are excellent methods to scale up from the lab-scale cell culture. Among the reactors, hollow fiber bioreactors contribute to a major part of high-density cell culture as they can provide a tremendous amount of surface area in a small volume for cell growth. As an alternative to hollow fiber reactors, a novel disposable bioreactor has been developed, which consists of a polymer-based supermacroporous material, cryogel, as a matrix for cell growth. Packed bed systems and disposable wave bioreactors have also been introduced for high cell density culture. These developments in high-density cell culture systems have led to the monoclonal antibody production in an economically favourable manner and made monoclonal antibodies one of the dominant therapeutic and diagnostic proteins in biopharmaceutical industry.