• 제목/요약/키워드: ion-exchange chromatography

검색결과 646건 처리시간 0.052초

아자크라운 에터를 포함한 이온교환수지에 의한 리튬 동위원소의 농축 (Enrichment of Lithium Isotope by an Ion Exchange Resin Containing Azacrown Ether)

  • 김동원;정용순;최기영;이용일;정영규;장영훈
    • 분석과학
    • /
    • 제10권6호
    • /
    • pp.403-407
    • /
    • 1997
  • Anchor group으로서 1,7,13-trioxa-4,10,16-triazacyclooctadecane($N_3O_3$)을 가지고 있는 이온교환수지를 사용하여 $^6Li$$^7Li$의 분리인자를 측정하였다. 가벼운 동위원소 $^6Li$는 액상에, 무거운 동위원소 $^7Li$는 이온교환수지상에 농축되었다. 2.0M 염화암모늄 용액을 용리액으로 사용한 컬럼 [0.9cm(내경)${\times}$20cm(높이)] 크로마토그래피에 의하여, 1.009의 분리인자, ${\alpha}$, $(^7Li/^6Li)_{resin}$/$(^7Li/^6Li)_{solution}$값을 얻었다. 이 값은 용리곡선과 동위원소비로부터 Glueckauf 이론을 적응하여 얻은 것이다.

  • PDF

Novel Purification Method of Kv 4.2 Potassium Channel from Rat Brain Membrane

  • Park, Sung-Soo
    • 대한의생명과학회지
    • /
    • 제18권2호
    • /
    • pp.96-103
    • /
    • 2012
  • Kv 4.2 ion channel protein has an ability to open at subthreshold membrane potentials and to recover quickly from inactivation. That is very important for neuronal signal transmission in vertebrate brain. In order to purify Kv 4.2 protein, the novel purification methods were experimented. The purification procedure utilized chromatography on DE-52 ion exchange column and affinity chromatography on a WGA-Sepharose 4B, and Kv 4.2 affinity column chromatography. It was found that 0.5% (wt./vol.) Triton X-100 detergent in lysis buffer worked well for Kv 4.2 protein solubilization from rat brain membrane. Protein quantitative determination was conducted by BCA method at 562 nm for each purification step to avoid determination interference of protein at 280 nm by detergent. The confirmation of Kv 4.2 existence and amount is performed using by SDS-PAGE/immunoblotting or 96-well dot blotting. The Kv 4.2 without interacting protein that contains carbohydrate, was purified from novel biochemical 3-steps purification method for further research.

고성능 액체크로마토그래피(HPLC)를 이용한 3가, 6가 크롬의 동시정량에 관한 연구 (Simultaneous Determination of Chromium (III) and Chromium(VI) by High Performance Liquid Chromatography(HPLC))

  • 노재훈;김치년;김춘성;김규상
    • 한국산업보건학회지
    • /
    • 제4권2호
    • /
    • pp.189-197
    • /
    • 1994
  • Analytic methods for Cr(VI) level in industrial hygienic field were suggested by the National Institute for Occupational Safety and Health(NIOSH method 7600, 7604). There were growing needs for measurement of Cr(III) and Cr(VI) levels simultaneously. Two analytical methods were suggested to determine Cr(III) and Cr(VI) levels simultaneously. The one is method by using reversed phase high peformance liquid chromatography(HPLC) and the other is by using ion exchange HPLC. The purpose of this work was to evaluate the usefulness of these two analytic methods. For the difference of ionic charges of Cr(III)-ethylendiamine tetraacetic acid(EDTA) chelate and $CrO_4{^-2}$, we could detect them simultaneously by ion exchange HPLC. Also, we attempted to determine the levels of Cr(III) and Cr(VI) chelated with sodium diethyldithiocarbamate(NaDDTC) by using reversed phase HPLC. The confirmation of Cr(III) and Cr(VI) were checked by fraction collector and nameless atomic absorption spectrometer. The optimal conditions for the formation of Cr(III)-EDTA chelate were two hours incubation period with pH 5. Cr(III)-EDTA and Cr(VI) in EDTA solution were successfully separated by anion exchange column using $Na_2CO_3/NaOH$ mixture as mobile phase. Peaks of Cr(III)-EDTA and Cr(VI) in EDTA were identified at 5 minutes and 7 minutes of retention time respectively by the ion exchange HPLC. The formation of Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates were twelve hours incubation period. Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates were separated by reversed phase column using methanol and water mixture as mobile phase. Peaks of Cr(VI)NaDDTC and Cr(III)-NaDDTC chelates were identified at 13 minutes and 26 minutes of retention time respectively by the reversed phase HPLC. Due to reduction of Cr(VI) to Cr(III), it seems to be not suitable for simultaneous determination of Cr(III)-NaDDTC and Cr(VI)-NaDDTC chelates by reversed phase HPLS. Simultaneos determination of Cr(III) and Cr(VI) by ion exchange HPLC was more accurate and simple method.

  • PDF

소(牛) 태아(胎兒) 대동맥(大動脈)으로부터 단백 분해효소 억제제 존제 하에 Lysyl Oxidase의 순수분리(純棒分離) -다양성(多形性)에 대(對)한 반론(反論)- (Purification and Characterization of Lysyl Oxidase from Fetal Bovine Aorta in the presence of protease inhibitors -Evidence against polymorphism-)

  • 한송
    • Journal of Oral Medicine and Pain
    • /
    • 제25권1호
    • /
    • pp.29-39
    • /
    • 2000
  • 소(牛) 태아 대동맥으로 부터 Urea 추출, Sephacryl S200 HR 크로마토그라피, Cibacron blue-agarose 친화 크로마토그라피, 그리고 Sephacryl S300 HR 크로마토그라피를 이용하여 lysyl oxidase를 순수분리를 하였고 분리가간 중 항상 단백분해 억제제를 첨가하였다. 순수 분리된 효소는 가교결합이 없는 교원단백질과 엘라스틴에 활성을 보였고, BAPN 같은 아미노나이트릴에 의하여 억제되였다. Sephacryl S300 HR 크로마토그라피로 분리 될 경우, 이 효소는 0.45의 $K_{av}$ ($V_t$의 65%)값을 보였고, 이온교환 고속액체 크로마토그라피의 경우에서는 이온 강도가 0.1-0.15 사이에서 하나의 피크로 용리되었다. 순수 분리된 이 효소는 SDS 폴리아크릴아마이드 전기영동에서는 하나의 밴드로 이동하였는데, 환원이 될 경우에는 분자량이 33,500, 비환원이 될 경우에는 분자량이 24,500의 위치로 이동하였다. 이온교환 고속액체 크로마토그라피의 결과를 참조하여, 다른 보고서와는 달리 소(牛) 태아(服兒) 대동맥(大動服)에는 여러 종류가 아닌 한 종류의 lysyl oxidase가 존재한다고 결론을 내렸다.

  • PDF

효모 배양액으로부터 재조합 모넬린의 정제와 특성 연구 (Purification and Characterization of Recombinant Monellin Produced from Yeast Culture Medium)

  • 김인호
    • KSBB Journal
    • /
    • 제13권5호
    • /
    • pp.535-539
    • /
    • 1998
  • The monellin, a sweet-taste protein, was expressed and secreted in Saccharomyces cerevisiae. The secreted menellin was concentrated using an ultrafiltration membrane with a nominal molecular weight cut off of 3,000 or by ammonium sulfate precipitation. The monellin was purified by G-25 gel filtration chromatography, followed by CM-Sepharose ion exchange chromatography. The purified monellin was characterized by SDS-PAGE (SDS-Polyacrylamide Gel Electrophoresis) and PHLC. The molecular weight of monellin was found to be 10,700 dalton, and its purity was over 95%.

  • PDF

L-PHA 렉틴의 분리 정제및 면역학적 연구 (Purification and Immunochemical Studies on L-PHA Lectin)

  • 정시련;서영아;소명숙;전경희
    • 약학회지
    • /
    • 제28권3호
    • /
    • pp.139-147
    • /
    • 1984
  • L-PHA, a lectin having lymphoagglutinating activity but devoid of erythroagglutinability which is contained in Korean white kidney bean (Phaseolus vulgaris L.), was isolated and purified through several techniques such as ion exchange chromatography, hydroxyapatite column and affinity chromatography. Purified L-PHA was identified as a single band by polyacrylamide disc gel electrophoresis. The molecular weight of the L-PHA was estimated about 125, 000 daltons by polyacrylamide gel electrophoresis and it was turned out having one subunit (probably dimer of the Yachnin's model) of Mr~60, 000. Immunochemical studies also tried and these results reconfirmed the purity of this L-PHA.

  • PDF

Real-time identification of the separated lanthanides by ion-exchange chromatography for no-carrier-added Ho-166 production

  • Aran Kim;Kanghyuk Choi
    • 대한방사성의약품학회지
    • /
    • 제7권2호
    • /
    • pp.69-77
    • /
    • 2021
  • No-carrier-added holmium-166 (n.c.a 166Ho) separation is performed based on the results of separation conditions using stable isotopes dysprosium (Dy) and holmium (Ho) to minimize radioactive waste from separation optimization procedures. Successful separation of two adjacent lanthanides was achieved by cation-exchange chromatography using a sulfonated resin in the H+ form (BP-800) and α-hydroxyisobutyric acid (α-HIBA) as eluent. For the identification process after separation of stable isotopes, the use of chromogenic reagents alternatively enables on-line detection because the lanthanides are hardly absorb light in the UV-vis region or exhibit radioactivity. Four different chromogenic reagents were pre-tested to evaluate suitable coloring reagents, of which 4-(2-Pyridylazo)resorcinol is the most recommendable considering the sensitivity and specificity for lanthanides. Lanthanide radioisotopes (RI) were monitored for separation with an RI detector using a lab-made separation LC system. Under the proper separation conditions, the n.c.a 166Ho was effectively obtained from a large amount of 100 mg dysprosium target within 2 hrs.

Amberlite IRA-900을 이용한 cyclodextrin glucotransferase의 최적 고정화 (Optimization of Cyclodextrin Glucanotransferase Immobilization on Amberlite IRA-900)

  • Seo, Hyo-Jin;Jung, Il-Hyong;Nam, Soo-Wan;Kim, Byung-Woo;Kim, Sung-Koo
    • 생명과학회지
    • /
    • 제14권5호
    • /
    • pp.794-799
    • /
    • 2004
  • Bacillus subtilis NAl/pKBl으로부터 생산된 cyclodextrin glucanotransferase (CGTase)는 cyclodextrin (CD)의 생산에 이용되었으며, 이에 사용된 CGTase는 ion-exchange chromatography와 gel filtration chromatography에 의해 정제되었다. 정제된 CGTase는 pH 6.0-7.0 범위, 60-$70^{\circ}C$에서 최대 활성을 나타내었으며, 다양한 이온결합성 고정화 담체를 이용하여 정제 효소의 고정화를 실시한 결과, 강염기성 음이온교환수지인 Amberlite IRA-900이 가장 우수한 고정화 효율을 나타내었다. 고정화된 효소는 pH 6.0, $60^{\circ}C$에서 최대 활성을 나타내었고, 그 활성이 약 1개월간 유지되어 cyclodextrin을 생산하기 위한 연속반응기내에서 장기간 사용이 가능함을 알 수 있었다.

디프테리아 toxin 정제와 무독화 toxoid 백신 생산 (Purification of Diphtheia Toxin and the Production of Detoxificated Toxoid Vaccine)

  • 조민;유연우
    • KSBB Journal
    • /
    • 제14권2호
    • /
    • pp.248-254
    • /
    • 1999
  • 본 연구에서는 디프테리아 toxin을 정제한 후 무독화하여 toxid를 생산하기 위하여 crude toxin을 2중 U.F를 통해 분자량 100.000이상의 단백질과 30.000이하의 배지 유래 단백질 및 색소를 제거한 결과 순도 1,300 Lf/mg PN의 toxin을 정제하였다. 이를 다시 DEAE-ion exehange chromatography를 통해 toxin을 정제한 후 무독화하여 순도 2,560 Lf/mg PN의 toxoid를 얻을 수 있었다. 이와 같이 생산된 디프테리아 toxoid는 동물 실험 결과 toxin으로 reversion이 발견되지 않았으며, 역가에 있어서도 crude toxin을 무독화한 후 정제한 toxoid와 비교하여 더 우수하였고 대한민국 생물학적 제제 기준에 규정된 성인용 디프테리아 백신 순도 기준 2,500 Lf/mg PN 이상에 적합하였다. 따라서 본 연구를 통해 성인용 디프테리아 백신의 생산 가능성을 확인하였다.

  • PDF