• Journal of Ginseng Research
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• 제22권4호
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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.

• 한국수정란이식학회지
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• 제33권4호
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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.

• 한국축산식품학회지
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• 제20권2호
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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.

• 한국식품영양학회지
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• 제13권2호
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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.

• 한국수산과학회지
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• 제48권3호
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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.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.351-354
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• 2005

한국산 개비자나무로부터 alkaloid계 항암 활성 물질인 Homoharringtonine의 분리 및 정제 공정을 개발하였다. 추출 용매로 메탄올을사용하여 biomass와 mthanol을 1:8의 비율로 분씩 3회 추출할 경우 개비자나무로부터 대부분(>99%)의 Homoharringtonine이 추출됨을 알 수 있었다. 흡착 공정에서는 활성 백토(active clay)를 사용하여 추출물에 포함되어 있는 식물유래 타르, 왁스 성분을 효과적으로 제거하였다. 크로마토그래피 공정을 통해서 52% 이상의 Homoharringtonine을 정제하였다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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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.

• BMB Reports
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• 제43권5호
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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.

• 생명과학회지
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• 제16권3호
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• pp.387-395
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• 2006

유도된 집파리유통 hemolymph중에서 Candida albicans의 3가지 anti-fungal peptides를 분리하였다. 3개 anti-fungal peptides는 분자량이 4-16 kDa 사이의 분명한 구별이 있을 뿐만 아니라, 각 peptide는 anti-fungal peptides작용이 있었다. 이들 peptide의 공통 특징은 모두 열을 받은 뒤 활성이 변하지 않는 비교적 강한 내열성을 보여주었다.

• KSBB Journal
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• 제15권4호
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• pp.342-345
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• 2000

식물세포배양으로부터 peroxidase를 대량 생산하기 위한 분리/정제 공정 으로서 세포를 파쇄하고 크로마토그래피 전처리로서 활성백토를 적용하였다. 활성백토는 미세한 세 포 조각 뿐만 아니라 여려가지 불순불을 선택적으로 흡착 하는 성질을 나타내었으며, 이를 통하여 효과적으로 정제 공정을 진행할 수 있었다 흡착을 통한 전처리 후 한외여 과장치를 이용하여 농축을 실시 하였으며, DEAE-Sepharose F FF를 이용한 크로마토그래피를 통해 정제가 이루어졌다. 활성을 나타내는 용액을 탈염, 농축 후 동결 건조하였다. 이 공정은 상업화에 필요한 대량 정제를 가능하게 하였으며 수율과 정제비용 측면에서 상당히 경제적인 공정임을 입증하였다, 활성백토를 이용한 흡착공정은 다른 효소 및 단백질의 경제적인 대량정제에 적용될 수 있으리라 기대된다.