• Title/Summary/Keyword: isolation and purification

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New Efficient Method for Isolation and Purification of Ginsenosides (Ginsenoside의 새로운 분리.정제 방법)

  • 김세원;황석연
    • Journal of Ginseng Research
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    • v.22 no.4
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    • pp.284-288
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    • 1998
  • This study was carried out to establish a new efficient method for isolation and purification of ginsenosides. Silica gel column chromatography, having been used for the isolation of ginsenosides, is advantageous to obtain a large amount of ginsenosides. However, it has a disadvantage to isolate ginsenosides to their highest purity. In addition, normal-or reverse-phase HPLC method thus far reported is confined to quantitative analysis. Especially, it has not been possible to isolate racemic 20(S)- and 20(R)-ginsenoside Rg2. In this experiment, isolation and purification of ginsenosides were accomplished by Diaion HP-20 adsorption chromatography, silica gel column chromatography, recrystalization and Prep. HPLC with or without Prep. TLC. From this study, we could establish a new efficient method for isolation and purification of 9 major and/or minor ginsenosides.

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Identification of a Technique Optimized for the Isolation of Spermatogonial Stem Cells from Mouse Testes

  • Han, Na Rae;Park, Hye Jin;Lee, Hyun;Yun, Jung Im;Choi, Kimyung;Lee, Eunsong;Lee, Seung Tae
    • Journal of Embryo Transfer
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    • v.33 no.4
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    • pp.327-336
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    • 2018
  • To date, there are no protocols optimized to the effective separation of spermatogonial stem cells (SSCs) from testicular cells derived from mouse testes, thus hindering studies based on mouse SSCs. In this study, we aimed to determine the most efficient purification method for the isolation of SSCs from mouse testes among previously described techniques. Isolation of SSCs from testicular cells derived from mouse testes was conducted using four different techniques: differential plating (DP), magnetic-activated cell sorting (MACS) post-DP, MACS, and positive and negative selection double MACS. DP was performed for 1, 2, 4, 8, or 16 h, and MACS was performed using EpCAM ($MACS^{EpCAM}$), Thy1 ($MACS^{Thy1}$), or GFR ${\alpha}1$ ($MACS^{GFR{\alpha}1}$) antibodies. The purification efficiency of each method was analyzed by measuring the percentage of cells that stained positively for alkaline phosphatase. DP for 8 h, $MACS^{Thy1}$ post-DP for 8 h, $MACS^{GFR{\alpha}1}$, positive selection double $MACS^{GFR{\alpha}1/EpCAM}$, and negative selection double $MACS^{GFR{\alpha}1/{\alpha}-SMA}$ were identified as the optimal protocols for isolation of SSCs from mouse testicular cells. Comparison of the purification efficiencies of the optimized isolation protocols showed that, numerically, the highest purification efficiency was obtained using $MACS^{GFR{\alpha}1}$. Overall, our results indicate that $MACS^{GFR{\alpha}1}$ is an appropriate purification technique for the isolation of SSCs from mouse testicular cells.

Isolation and Purification of Lactoferrin from Korean Native Cow's Colostrum (한우 초유로부터 Lactoferrin의 분리.정제)

  • 양희진;하월규;양동훈;박기문;이수원
    • Food Science of Animal Resources
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    • v.20 no.2
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    • pp.125-132
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    • 2000
  • Lactoferrin was isolated from the colostrum of Korean native cow by using several purification steps such as batch extraction, ion exchange chromatography, gel filtration chromatography and affinity chromatography. Other whey protein components that having similar molecular weight and affinity to lactoferrin were gradually removed from crude Korean native cow's lactoferrin during the purification steps. The amount of lactoferrin collected from a liter of Korean native cow's colostrum was 65mg and the recovery rate was 29.4%. The molecular weight of the purified Korean native cow's lactoferrin was estimated approximately 81,000dalton.

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Isolation and Purification of Antimicrobial Active Substances from Aloe vera peel(II) (Aloe vera peel에서 항균활성 물질의 분리.정제(II))

  • 박정순;신용서;류일환;이갑상
    • The Korean Journal of Food And Nutrition
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    • v.13 no.2
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    • pp.146-151
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    • 2000
  • To evaluate anticaries and antiinflammation of Aloe vera peel, antimicrobial substances were extracted from Aloe vera peel and identified. The antimicrobial active substances of water extract were successfully purified with solvent fractionation, silica gel column chromatography, preparative thin layer chromatography and UV spectrophotometer. Two purified active substances were identified as aloe-emodin and barbaloin by Mass Spectrometer, 1H-NMR and FT-IR.

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Isolation of Eckol from Ecklonia cava via Centrifugal Partition Chromatography (CPC) and Characterization of it's Anti-inflammatory Activity (고속원심분배 크로마토그래피를 이용한 감태(Ecklonia cava)로부터 Eckol의 분리 및 항염증 활성)

  • Kim, Yoon Taek;Lee, Ji-Hyeok;Ko, Ju-Young;Oh, Jae-Young;Lee, Won-U;Sok, Chang Hyun;Hong, Jin Tae;Jeon, You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.3
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    • pp.301-307
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    • 2015
  • Phlorotannins and marine algal polyphenols, including dieckol, 6,6-bieckol, phloroglucinol, phlorofucofuroeckol-A, and eckol, were isolated from brown seaweeds. These compounds have beneficial bioactivities, and Ecklonia cava has become widely used for the extraction and isolation of phlorotannins. Eckol, in particular, has been to shown to have antioxidant, anti-inflammatory, anticoagulatory, and photoprotective properties. However, due to its low abundance in weaweed, the isolation and purification of eckol are difficult. Its limited availability renders the isolation and purification of eckol labor-intensive processes. Centrifugal partition chromatography (CPC) is an efficient technique for the isolation and purification of eckol. In this study, eckol was isolated from the ethyl acetate fraction of the 70% ethanol extract of E. cava using CPC with a two-phase solvent system of a n-hexane:EtOAc:methanol:water (2:8:3:7, v/v) solution. The purity and anti-inflammatory activity of the isolated eckol were verified by high-performance liquid chromatography and by assaying lipopolysaccharide-induced inflammatory responses in an immortalized murine BV2 microglial cell line, respectively. In conclusion, CPC is a useful technique for simple and efficient isolation of eckol from E. cava.

Purification and Characterization of Homoharringtonine from Cephalotaxus koreana

  • Kim, Byung-Sik;Kim, Jin-Hyun
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.351-354
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    • 2005
  • A new isolation and purification method was developed aiming at increasing yield and purity for homoharringtonine. This method was a simple andefficient procedures, for the isolation and purification of homoharringtonine from Cephalotaxus koreana, consisting of solvent extraction, adsorbent treatment, low-pressure chromatography, and high performance liquid chromatography (HPLC). The crude homoharringtonine was efficiently pre-purified adequately to perform HPLC through a combination withadsorbent treatment and low-pressure chromatogaphy. The homoharringtonine can be simply obtained with high purity and yield from crude homoharringtonine by HPLC. Purified homoharringtonine was characterized.

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A Novel Purification Process for Homoharringtonine from Celphalotaxus koreana

  • Sung, Ju-Li;Kim, Jin-Hyun
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.521-524
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    • 2003
  • An effective purification method was developed for producing Homoharringtonine (HHT), to guarantee high purity and yield from Cephalotaxus koreana. This process was a simple and efficient procedure, for the isolation and purification of HHT form the biomass of Cephalotaxus koreana, consisting of extract, adsorbent treatment, precipitation and followed by a chromatography. The extraction, adsorbent treatment and precipitation in pre-purification process allows for rapid and efficient separation of HHT from many compound and dramatically increases the yield and purity of crude HHT for HPLC purification steps compared to alternative processes. This purification processes serves to minimize solvent usage, size, and complexity of the operations for HHT purification.

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Retrospective analyses of the bottleneck in purification of eukaryotic proteins from Escherichia coli as affected by molecular weight, cysteine content and isoelectric point

  • Jeon, Won-Bae
    • BMB Reports
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    • v.43 no.5
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    • pp.319-324
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    • 2010
  • Experimental bioinformatics data obtained from an E. coli cell-based eukaryotic protein purification experiment were analyzed in order to identify any bottleneck as well as the factors affecting the target purification. All targets were expressed as His-tagged maltose-binding protein (MBP) fusion constructs and were initially purified by immobilized metal affinity chromatography (IMAC). The targets were subsequently separated from the His-tagged MBP through TEV protease cleavage followed by a second IMAC isolation. Of the 743 total purification trials, 342 yielded more than 3 mg of target proteins for structural studies. The major reason for failure of target purification was poor TEV proteolysis. The overall success rate for target purification decreased linearly as cysteine content or isoelectric point (pI) of the target increased. This pattern of pI versus overall success rate strongly suggests that pI should be incorporated into target scoring criteria with a threshold value.

Isolation and Purification of Novel Anti-Fungal Peptides from Hemolymph of Immunized Larvae of Housefly, Musca domestica (집파리유충 hemolymph 중신형의 anti-fungal peptides의 분리정제)

  • Gu Li-Juan;Wu Jian-Wei;Su Xiao-Qing;Sung Chang-Keun
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.387-395
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    • 2006
  • To isolate and purify anti-fungal active substances from immunized housefly (Musca domestica), low dose of Candida albicans was injected into the larvae of the housefly to induce the appearance of potent anti-fungal active substances in the hemolymph. This purification work was performed by the routine isolation and purification processes of protein, namely, solid phase extraction (SPE), SDS-PACE electrophoresis, HPLC purification. Three 4-16 kDa peptides which exhibited antifungal activity against Candida albican and other fungi were isolated from induced hemolymph. Consequently, further anti-fungal activity study showed that these three peptides were different either in molecular weight or in anti-fungal activity. All isolated substances were proved to be active and resistant to high-temperature. It was deduced that these peptides isolated from induced housefly were novel members of the insect defensin family and they were inducible.

A New large-scale Pre-purification for Peroxidase from Plant Cell Cultures (식물세포 배양으로부터 Peroxidase 대량 정제를 위한 전처리 공정 개발)

  • 표상현
    • KSBB Journal
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    • v.15 no.4
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    • pp.342-345
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    • 2000
  • A novel pre-purification method was developed for producing peroxidase to guarantee high purity and yield from plant cell cultures in large-scale process. This method was a simple and efficient procedure for the isolation and pre-purification of peroxidase from the biomass consisting of active clay treatment followed by cationic exchange chromatography. The use of active clay in the pre-purification process allows for rapid and efficient separation of peroxidase from interfering compounds and dramatically increases yield and purity of crude peroxidase for purification steps compared to alternative processes. This pre-purification process serves to minimize the buffer usage size and complexity of the HPLC operations for peroxidase purification. This process is readily scalable to a pilot plant and eventually to a production environment where mass production of material are expected to be produced.

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