• Title/Summary/Keyword: Capillary Electrophoresis

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Sheathless electrospray ionization with integrated metal emitter on microfluidic device (전기 분무 이온화를 이용한 단백질 질량분석용 마이크로 유체 소자의 제작 및 실험)

  • Kim, Min-Su;Joo, Hwang-Soo;Lee, Kook-Nyung;Kim, Byung-Gee;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 2004.07c
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    • pp.2102-2104
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    • 2004
  • In this study, sheathless electrospray from PDMS/glass microchips with conducting metal emitter tip is described. A chip-based capillary electrophoresis/mass spectrometry (CE/MS) system has advantages of the CE separation and on-line electrospray detection of peptide solution. We have fabricated a new electrospray ionization(ESI) device composed of the metal emitter tip and CE separation channel monolithically in a glass microchip. The separation channel and metal emitter tip are fabricated using a glass wet etching and gold electro plating process, respectively. The fabricated micro electrospray chip was tested by spraying peptide sample for mass spectrometric analysis. Singlely-charged peak and doublely-charged peak of peptide were detected and further MS/MS fragmentation was performed in each peak. Direct comparisons with conventional glass or fused silica emitters showed very similar performance with respect to signal strength and stability.

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Microfabricated Disposable Microchip with a Capillary Electrophoresis and Integrated Amperometric Detection (모세관 전기영동 및 전기화학적 검출 시스템을 위한 일회용 마이크로칩)

  • Kim, Ju-Ho;Kang, C.J.;Kim, Yong-Sang
    • Proceedings of the KIEE Conference
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    • 2004.07c
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    • pp.2131-2133
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    • 2004
  • 모세관 전기영동 및 전기화학적 검출 시스템을 마이크로 시스템에 적용하여 ITO 유리기판과 polydimethylsiloxane (PDMS)로 제작하였다. 제작된 모세관 전기영동 및 전기화학적 검출 시스템은 일회용으로 사용가능하며 전기화학적 검출에 아주 적합한 특성을 보인다. 모세관 전기영동 및 전기화학적 검출 시스템은 주입과 분리 채널 (80 ${\mu}m$${\sim}$ 40 ${\mu}m$ 깊이)을 가진 PDMS 층과 유리기판 위에 검출 전극으로 사용되는 ITO가 형성된 층으로 구성된다. PDMS 층과 ITO 유리 기판은 UV-$O_3$ cleaner를 사용하여 접합하였다. 완충용액은 10 mM 2-(N-morpholino)ethanesulfonic acid (MES)를 사용하였고 분석물질은 1 mM 농도의 dopamine과 1 mM 농도의 catechol을 사용하였다. 60 V/cm 전계로 주입 및 분리를 하였으며 작업전극과 기준전극 간의 전위는 +600 mV로 유지하며 분석물질의 농도에 비례하는 전류량법으로 측정하였다. 전기화학적 검출 회로는 천기영동 전계의 간섭으로부터 분리하였다. 10 mM MES 완충용액에서 바탕 전류의 크기가 ${\sim}$10 pA 일 때 측정전류 값은 10 nA이다. 측정된 피크 값은 기존의 Au 전극과 비교하여 선택성, 감도, 분해능이 유사한 특성을 보여준다.

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모세관 전기이동을 이용한 ${\delta}$-Aminolevulinc acid와 Porphobilinogen의 분리

  • Kim, Jin-Nam;Yun, Jong-Seon;Ryu, Hwa-Won
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.512-515
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    • 2000
  • In this study, ${\delta}-aminolevulinic$ acid, porphobilinogen, glycine and levulinic acid were succesfully separated by capillary electrophoresis(CE). We established the separative conditions of the mixture containing four components by CE. The borate buffered solution was used for CE electrolyte, and its pH was adjusted to $9.25{\sim}9.42$. Under constant current or constant voltage, higher concentraion of borate produced better resolution of four components, but adversely affected migration rates , resulting in longer analysis time. While migration time was faster with increase in applied voltage, but adversely affected resolution. Each component was separated well in borate buffer of 30mM at the applied voltage of 20kV.

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Validation of 17 Microsatellite Markers for Parentage Verification and Identity Test in Chinese Holstein Cattle

  • Zhang, Yi;Wang, Yachun;Sun, Dongxiao;Yu, Ying;Zhang, Yuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.4
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    • pp.425-429
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    • 2010
  • To develop an efficient DNA typing system for Chinese Holstein cattle, 17 microsatellites, which were amplified in four fluorescent multiplex reactions and genotyped by two capillary electrophoresis injections, were evaluated for parentage verification and identity test. These markers were highly polymorphic with a mean of 8.35 alleles per locus and an average expected heterozygosity of 0.711 in 371 individuals. Parentage exclusion probability with only one sampled parent was approximately 0.999. Parentage exclusion probability when another parent' genotype was known was over 0.99999. Overall probability of identity, i.e. the probability that two animals share a common genotype by chance, was $1.52{\times}10^{-16}$. In a test case of parentage assignment, the 17 loci assigned 31 out of 33 cows to the pedigree sires with 95% confidence, while 2 cows were excluded from the paternity relationship with candidate sires. The results demonstrated the high efficacy of the 17 markers in parentage analysis and individual identification for Chinese Holstein cattle.

Serological Detection of Plant Viruses with latex-test (Latex-Test에 의한 식물 바이러스의 검정)

  • 박은경;김정화;이영근
    • Journal of the Korean Society of Tobacco Science
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    • v.1 no.1
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    • pp.33-38
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    • 1979
  • A simple test using latex-agglutination was developed to detect serologically tobacco mosaic virus (TMV), soybean mosaic virus (SoyMV), and barley stripe mosaic virus (BSMV) from infected Plants. Latex spheres ( 0.81 $\mu$, Difco) were adsorbed with immuno globulin purified by electrophoresis from crude antiserum against viruses. The antibody- sensitized latex suspension was mixed with sap from virus -infected leaves in a glass capillary tube (inner diam. 1mm $\times$ 100 mm length) The mixture, after agitation, was observed under a stereo microscope at low magnification (X20 - X4O), to examine the reaction between antigen (Virus) and its antibody. Flocculation occurred when the reaction was positive.

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Detection of Phagocytosis-Promoting Factor of Culture Supernatant from Feline Peripheral Blood Mononuclear Cells Cultured with Egg White Derivatives (계난백유래물질로 배양한 고양이 말초혈액 단핵구세포 배양상층액중의 탐식촉진인자 검출)

  • 양만표;김기홍
    • Journal of Veterinary Clinics
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    • v.16 no.1
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    • pp.31-36
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    • 1999
  • The aim of this study is to determine the phagocytosis-promoting factor(s) for feline peripheral blood polymorphonuclear cells (PMN) by culture supernatant from mono-nuclear cells (MNC) treated with egg white derivatives (EWD). The phagocytic activity of PMN was analyzed by a flow cytometry system. The EWD did not show direct effect on the phagocytic response of PMN. The phagocytic activity of PMN was enhanced by culture supernatant from MNC but not PMN treated with EWD. Therefore, it was suggested that the enhanced phagocytic activity of feline PMN could be mediated by humoral factor(s) released from MNC treated with EWD. Thus, the phagocytosis-promoting factor(s) in supernatant fraction from MNC culture treated with EWD were isolated by reverse phase high pressure liquid chromatography. The resulting supernatant fraction on 29.02 minutes of retention time showed high phagocytic activity of PMN. The molecular weight of this supernatant fraction was 16 to 18 kDa when analyzed by capillary electrophoresis. The isoelectric point was pH 5.76 when assessed by ion-exchange chromatography. These results suggest that EWD stimulates feline MNC to elaborate a phagocytosis-promoting factor, 16 to 18 kDa of molecular weight, which could be an important mediator for the enhancement of phagocytic activity of feline peripheral blood phagocytes. Further study will be needed to elucidate this phagocytic factor.

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Acinetobacter calcoaceticus Glucose-1-phosphate Thymidylyltransferase: Cloning, Sequencing, and Expression in E.coli

  • Eun, Suk-Ho;Kim, Dae-Jin;Kim, Yu-Sam
    • BMB Reports
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    • v.34 no.3
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    • pp.230-236
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    • 2001
  • dTDP-rhamnose is synthesized from dTTP and glucose-1-phosphate by four enzymatic steps in the gram-negative bacteria. By using a homologous PCR product, a gene cluster encoding four genes (rfbA, rfbB, rfbC, rfbD) involved in L-rhamnose biosynthesis by Acinetobacter calcoaceticus was isolated and sequenced. The four genes were clustered on the biosynthetic operon in the order of rfbB, D, A, C. A gene, rfbA, encoding glucose-l-phosphate thymidylyltransferase (RfbA), was cloned from A. calcoaceticus pathogenic and encapsulated in the gram-negative bacterium. This enzyme catalyzes the formation of dTDP-D-glucose From $\alpha$-D-glucose-1-phosphate and dTTP.RfbA was amplified by PCR and inserted into the $T_7$ expression system. The activity of RfbA was determined by the capillary electrophoresis. The $K_m$ values for dTTP and $\alpha$-D-glucose-1-phosphate were calculated to be 1.27 mM and 0.80 mM, respectively by using the Line-Weaver Burk plot. RfbA is inactivated by diethylpyrocarbonate.

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Enantioselective Pharmacokinetics of Carvedilol in Human Volunteers

  • Phuong, Nuyen-Thi;Lee, Beam-Jin;Choi, Jung-Kap;Kang, Jong-Seong;Kwon, Kwang-il
    • Archives of Pharmacal Research
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    • v.27 no.9
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    • pp.973-977
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    • 2004
  • Carvedilol is administered as a racemic mixture of the R(+)- and S(-)-enantiomers, although they exhibit different pharmacological effects. To investigate the stereoselective pharmacoki-netics, the enantiomeric separation of carvedilol in human plasma was undertaken using capil-lary electrophoresis (CE). Resolution of the enantiomers was achieved using 2-hydoxypropyl-$\beta$-cyclodextrin as the chiral selector. Phosphate buffer (50 mM, pH 4.0) containing 10 mM of 2-hydoxypropropyl-$\beta$-cyclodextrin was used as electrolytic buffer. Achiral separation was carried out with the same electrolytic buffer without chiral selector. Following a single oral administra-tion of 25-mg carvedilol to 11 healthy, male volunteers, stereoselective pharmacokinetic analy-sis was undertaken. The maximum plasma concentrations ( $C_{max}$) were 48.9 and 21.6 ng/mL for (R)-carvedilol and (S)-carvedilol, respectively, determined by the chiral method. The profiles of the plasma concentration of (RS)-carvedilol showed $C_{max}$ of 71.5, 72.2, and 73.5 ng/mL, as determined by the CE, HPLC/FD methods and calculations from the data of the chiral method, respectively.y.y.

Chemical Analysis of Water Soluble Aerosols at Kosan, Cheju lsland (제주도 고산에서 수용성 에어로졸의 화학적 성분 분석)

  • 이호근;박경윤;서명석;장광미;강창희;허철구
    • Journal of Korean Society for Atmospheric Environment
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    • v.11 no.3
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    • pp.245-252
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    • 1995
  • Atmospheric aerosols were collected by a High Volume Tape Sampler from March 1992 to December 1993 ar Korean, Cheju, Korea. The water soluble ion concentrations in aerosol were analyzed. The concentrations of cations (N $a^{+}$, $K^{+}$, $Ca^{2+}$, $Mg^{2+}$, N $H_{4}$$^{+}$) were determined by an Inductively Coupled Plazma(ICP) or an Atomic Absorption Spectrometer(AAS), and those of anions (C $l^{[-10]}$ , N $O_{3}$$^{[-10]}$ , S $O_{4}$$^{2-}$) were analayzed by the capillary electrophoresis method. The $Ca^{2+}$, S $O_{4}$$^{2-}$ and N $O_{3}$$^{[-10]}$ concentrations in spring were higher than those in other seasons. The lowest concentrations of these elements were found in summer, largely due to scavenging by frequent rains. Especially the $Ca^{2+}$ concentration on April was three to four times higher than the annual mean concentration. The enrichment factor(E.F.) of each element was calculated. The annual mean E.F. values of the $Ca^{2+}$, $Mg^{2+}$ and C $l^{[-10]}$ in 1992 were the same as those in 1993 except $k^{+}$ and S $O_{4}$$^{2-}$. The correlation formula between all cations and anions for the whole period was Anions = 0.759 * Cations + 0.066.Cations + 0.066.

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Microbial Detection and Identification Using Biosensors

  • Kim, Sol
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2008.05a
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    • pp.135-135
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    • 2008
  • Various biosensors were evaluated for identifying and detecting foodborne pathogens in a rapid and effective manner. First, five strains of Escherichia coli and six strains of Salmonella were identified using Fourier transform infrared spectroscopy and a statistical program. For doing this, lipopolysaccharides (LPSs) and outer membrane proteins (OMPs) were extracted from a cell wall of each bacterial strain. As a result, each strain was identifed at the level of 97% for E. coli and 100% for Salmonella. Second, E. coli O157:H7, S. Enteritidis, and Listeria monocytogenes were identified by multiplex PCR products from four specific genes of each bacteria using a capillary electrophoresis (CE). Also, ground beef for E. coli O157:H7, lettuce for S. Enteritidis, and hot dog for L. monocytogenes were used to determine the possibility of detecting pathogens in foods. Foods inoculated with respective pathogen were cultivated for six hours and multiplex PCR products were obtained and assessed. The minimum detection levels of tested bacteria were <10 cells/g, <10 cells/g, and $10^4$ cells/g for E. coli O157:H7, S. Enteritidis, and L. monocytogenes, respectively. Third, it was possible to detect S. Typhimurium in a pure culture and lettuce by a bioluminescence-based detection assay using both recombinant bacteriophage P22::luxI and a bioluminescent bioreporter. In addition, bacteriophage T4 was quantitatively monitored using E. coli including luxCDABE genes.

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