• Bulletin of the Korean Chemical Society
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• 제32권12호
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• pp.4253-4257
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• 2011

Sodium dodecyl sulfate-capillary gel electrophoresis (SDS-CGE) and capillary isoelectric focusing (CIEF) were employed to characterize and compare ricin E purified from the small grain seeds of Ricinus communis with ricin D isoform. During the purification of ricin E using ion-exchange and size-exclusion chromatography, SDS-CGE was found to be useful for monitoring the efficiencies of purification steps. SDS-CGE showed that the molecular size of ricin E was not significantly different from that of ricin D, which was confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. CIEF was useful for discriminating ricin E from ricin D based on their isoelectric points (pI). The pI values of ricin E and D were 8.6-8.8 and 7.0-7.4, respectively. This study demonstrates the usefulness of SDS-CGE and CIEF for the characterization of ricin toxins.

• Bulletin of the Korean Chemical Society
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• 제24권7호
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• pp.943-947
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• 2003

We have studied the migration behavior of single-stranded DNA using capillary gel electrophoresis under various conditions. It was found that optimum electric fields should be less than 150 V/cm for the good tradeoff between the separation time and the resolution. It seems that the gel matrix with the combination of different polymer average molecular weights is important to extend the maximum readable DNA bases. The total gel concentration less than 3.1% in the mixed gel system showed good separation efficiency up to 600 bases. The best result was obtained with the poy(ethylene)oxide (PEO) gel concentration of 1.2% of Mr 8,000,000 and 1.8% of Mr 600,000. We observed that the capillary length between 50 cm to 100 cm (effective length) should be employed for the optimization between the total DNA migration time and the maximum readable length. A trizma base-boric acid-ethlyenediaminetetraacetic acid (EDTA) (TBE) buffer was commonly used for DNA sequencing, but we found that 3-[tris(hydroxymethyl)methyl amino]-1-propane sulfonic acid (TAPS) buffer worked as well for the single-stranded DNA separation. Especially, TAPS buffer showed a good resolution for very short DNA bases (1 to 30 bases).

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.230.3-231
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• 2003

The therapeutic use of protein pharmaceuticals produced by recombinant DNA technology is increasing in recent decades. In order to investigate the quality of recombinant proteins, it is important to identify and assign the impurities produced in the process of recombination or in storage conditions. Capillary Electrophoresis is emerging technology exhibiting high sensitivity, selectivity and speed and may be most powerful tools for this application. In this study, human growth hormone (hGH) has been analyzed by various mode of capillary electrophoresis such as capillary zone electrophoresis (CZE), capillary gel electrophoresis (CGE), and capillary isoelectric focusing (cIEF) to indicate the chemically or biologically oriented variants and the degraded fragments. (omitted)

• Food Science and Biotechnology
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• 제18권4호
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• pp.823-832
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• 2009

In recent years, special concerns have been raised about the safety assessment of foods and food ingredients derived from genetically modified organisms (GMOs). A growing number of countries establish regulations and laws for GMOs in order to allow consumers an informed choice. In this case, a lot of methods have been developed for the detection of GMOs. However, the reproducibility among methods and laboratories is still a problem. Consequently, it is still in great demand for more effective methods. In comparison with the gel electrophoresis, the capillary electrophoresis (CE) technology has some unique advantages, such as high resolution efficiency and less time consumption. Therefore, some CE-based methods have been developed for the detection of GMOs in recent years. All kinds of CE detection methods, such as ultraviolet (UV), laser induced fluorescence (LIF), and chemiluminescence (CL) detection, have been used for GMOs detection. Microchip capillary electrophoresis (MCE) methods have also been used for GMOs detection and they have shown some unique advantages.

• Bulletin of the Korean Chemical Society
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• 제30권11호
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• pp.2655-2660
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• 2009

A breed of cattle, i.e., Korean cattle (Hanwoo), was identified based on the DNA mobilities of their microsatellites (MSs) by capillary gel electrophoresis (CGE) with a laser-induced fluorescence (LIF) detector. The MS markers were used for the accurate identification of species-specific genes. The DNA mobilities of the MS markers of Hanwoo and Holstein were measured using a CGE system with a fused-silica capillary (inner diameter of 75 ${\mu}m$, outer diameter of 365 ${\mu}m$, and total length of 50 cm). The capillary was dynamically coated with 1.0% (w/v) polyvinylpyrrolidone ($M_r$ = 1,000,000) and then filled with a mixture of 1.3% (w/v) poly(ethylene oxide) ($M_r$ = 600,000) and 1.9% (w/v) poly(ethylene oxide) (Mr = 8,000,000) as a sieving gel matrix. The species-specific genes of Hanwoo and Holstein were clearly distinguished within 33 min. This CGE assay technique is expected to be a useful analytical method for the fast and accurate identification of breeds of cattle.

• BMB Reports
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• 제31권4호
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• pp.384-390
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• 1998

DNA fragments (51-587 bp) were separated by capillary electrophoresis using entangled polymer, hydroxyethylcellulose, in uncoated fused silica capillary columns. The factors affecting the separation of DNA fragments with hydroxyethylcellulose media were evaluated, i.e., the concentration of buffer and entangled polymer, effects of additives (methanol, ethidium bromide, EDTA), temperature, and injection methods. Maximum performance was obtained by adding 5% methanol in 0.5% hydroxyethylcellulose solution at $30^{\circ}C$. Addition of methanol in polymer media increased the resolution of small size DNA fragments (< 100 bp). On the other hand, addition of ethidium bromide and EDTA, which are commonly used in conventional DNA separation, reduced the resolution of DNA fragments in the polymer solution. It turns out that the separation behavior of DNA in entangled polymer is more sensitive to the running condition compared to that in polyacrylamide gel-filled capillary, but the reproducibility of DNA separation in entangled polymer is reliable.

• Mass Spectrometry Letters
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• 제3권1호
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• pp.10-14
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• 2012

Artificially oxidized cysteine residues in peroxiredoxin 6 (Prx6) were detected by electrospray interface capillary liquid chromatography-linear ion trap mass spectrometry after the preparation of two-dimensional gel electrophoresis (2D-GE). We used Prx6 as a model protein because it possesses only two cysteine residues at the 47th and 91st positions. The spot of Prx6 on 2D-GE undergoes a basic (isoelectric point, pI 6.6) to acidic (pI 6.2) shift by exposure to peroxide due to selective overoxidation of the active-site cysteine Cys-47 but not of Cys-91. However, we detected a tryptic peptide containing cysteine sulfonic acid at the 47th position from the basic spot and a peptide containing both oxidized Cys-47 and oxidized Cys-91 from the acidic spot of Prx6 after the separation by 2D-GE. We prepared two types of oxidized Prx6s: carrying oxidized Cys-47 (single oxidized Prx6), and other carrying both oxidized Cys-47 and Cys-91 (double oxidized Prx6). Using these oxidized Prx6s, the single oxidized Prx6 and double oxidized Prx6 migrated to pIs at 6.2 and 5.9, respectively. These results suggest that oxidized Cys-47 from the basic spot and oxidized Cys-91 from the acidic spot are generated by artificial oxidation during sample handling processes after isoelectric focusing of 2D-GE. Therefore, it is important to make sure of the origin of cysteine oxidation, if it is physiological or artificial, when an oxidized cysteine residue(s) is identified.

• 한국동물위생학회지
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• 제34권3호
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• pp.227-234
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• 2011

Brucella (B.) abortus is a facultative intracellular pathogen that infects a wide variety of animal species and human. Brucellosis is the zoonosis and an extremely important disease around the world. Although the eight species can be differentiated by conventional phenotypic tests, these species display a high degree of DNA homology in DNA-DNA hybridization assay (>90%). Various methods have been established for genotyping of Brucella species, but most of analytical methods are lack reproducibility and limited capability to differentiate them. The attempt of this study was to evaluate multiple-locus VNTR analysis (MLVA) for use of epidemiological trace-back analysis in bovine brucellosis. Ninety-four B. abortus isolates from Gyeongbuk province during 2006~2010 were analyzed using 16 VNTR locus. High resolution automatic capillary electrophoresis system was used for more throughput, simpleer, faster, and better discriminable than conventional gel electrophoresis. As a result, 13 different genotypes were identified from 94 B. abortus isolates. MLVA could contribute to epidemiological trace-back analysis of bovine brucellosis.

• Bulletin of the Korean Chemical Society
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• 제29권1호
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• pp.153-158
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• 2008

A microchip-based capillary gel electrophoresis (MCGE) technique was developed for the ultra-fast detection and differentiation of Candidatus Mycoplasma haemominutum (Candidatus M. haemominutum, California strain) and Mycoplasma haemofelis (M. haemofelis, Ohio strain) in Korean feral cats through the application of programmed field strength gradients (PFSG) in a conventional glass double-T microchip. The effects of the poly (ethyleneoxide) (PEO) concentration and electric field strength on the separation of DNA fragments were investigated. The PCR-amplified products of Candidatus M. haemominutum (202-bp) and M. haemofelis (273-bp) were analyzed by MCGE within 75 s under a constant applied electric field of 117.6 V/cm and a sieving matrix of 0.3% PEO (Mr 8 000 000). When the PFSG was applied, MCGE analysis generated results 6.8-times faster without any loss of resolution or reproducibility. The MCGE-PFSG technique was also applied to eleven samples selected randomly from 33 positive samples. The samples were detected and differentiated within 11 s. The analysis time of the MCGE-PFSG technique was approximately 980-times faster than that using conventional slab gel electrophoresis.

• 대한약학회:학술대회논문집
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• 대한약학회 2000년도 춘계총회 및 학술대회
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• pp.196.3-197
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• 2000