• 제목/요약/키워드: Purification method

검색결과 848건 처리시간 0.027초

Purification of Filamentous Bacteriophage M13 by Expanded Bed Anion Exchange Chromatography

  • Tau Chuan;Chee Kin;Wen Siang;Beng Ti;Wan, Wan-Mohammad;Arbakariya
    • Journal of Microbiology
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    • 제42권3호
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    • pp.228-232
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    • 2004
  • In this paper, we investigated the development of a simplified and rapid primary capture step for the recovery of M13 bacteriophage from particulate-containing feedstock. M13 bacteriophage, carrying an insert, was propagated and subsequently purified by the application of both conventional multiple steps and expanded bed anion exchange chromatography. In the conventional method, precipitation was conducted with PEG/NaCl, and centrifugation was also performed. In the single step expanded bed anion exchange adsorption, UpFront FastLine$\_$TM/20 (20mm i.d.) from UpFront Chromatography was used as the contactor, while 54$m\ell$ (H$\_$o/=15cm) of STREAMLINE DEAE (p=1.2 g/㎤) from Amersham Pharmacia Biotechnology was used as the anion exchanger. The performance of the two methods were evaluated, analysed, and compared. It was demonstrated that the purification of the M13 bacteriophage, using expanded bed anion exchange adsorption, yielded the higher recovery percentage, at 82.86%. The conventional multiple step method yielded the lower recovery percentage, 36.07%. The generic application of this integrated technique has also been assessed.

A new purification method for the Fab and F(ab)2 fragment of 145-2C11, hamster anti-mouse CD3ε antibody

  • Kwack, Kyu-Bum
    • BMB Reports
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    • 제33권2호
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    • pp.188-192
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    • 2000
  • Recombinant protein G has been utilized in the purification of antibodies from various mammalian species based on the interaction of antibodies with protein G. The interaction between immunoglobulin and protein G may not be restricted to the Fc protion of antibodies, as many different $F(ab)_2$ or Fab fragments can also bind to protein G. I found both FAb $F(ab)_2$ of 145-2C11, a hamster anti-mouse $CD3{\varepsilon}$ antibody, bound to the protein G-sepharose. Interestingly, Fab and $F(ab)_2$ of 145-2C11 did not bind to the protein A-sepharose. The binding of Fab and $F(ab)_2$ of 145-2C11 to protein G provided a useful method to remove proteases, chopped fragments of the Fc region, and other contaminating proteins. The remaining intact antibody in the protease reaction mixture can be removed by using a protein A-sepharose, because the Fab and $F(ab)_2$ portions of 145-2C11 did not bind to protein A-sepharose. The specific binding of Fab and $F(ab)_2$ portions of 145-sC11 to a protein G-sepharose (though not to a protein A-sepharose) and binding of intact 145-2C11 to both protein A- and G-sepharose will be useful in developing an effective purification protocol for Fab and $F(ab)_2$ portions of 145-2C11.

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Improved recovery of active GST-fusion proteins from insoluble aggregates: solubilization and purification conditions using PKM2 and HtrA2 as model proteins

  • Park, Dae-Wook;Kim, Sang-Soo;Nam, Min-Kyung;Kim, Goo-Young;Kim, Jung-Ho;Rhim, Hyang-Shuk
    • BMB Reports
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    • 제44권4호
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    • pp.279-284
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    • 2011
  • The glutathione S-transferase (GST) system is useful for increasing protein solubility and purifying soluble GST fusion proteins. However, purifying half of the GST fusion proteins is still difficult, because they are virtually insoluble under non-denaturing conditions. To optimize a simple and rapid purification condition for GST-pyruvate kinase muscle 2 (GST-PKM2) protein, we used 1% sarkosyl for lysis and a 1 : 200 ratio of sarkosyl to Triton X-100 (S-T) for purification. We purified the GST-PKM2 protein with a high yield, approximately 5 mg/L culture, which was 33 times higher than that prepared using a conventional method. Notably, the GST-high-temperature requirement A2 (GST-HtrA2) protein, used as a model protein for functional activity, fully maintained its proteolytic activity, even when purified under our S-T condition. This method may be useful to apply to other biologically important proteins that become highly insoluble in the prokaryotic expression system.

Characterization of a Korean Isolate of Dasheen mosaic virus Isolated from Taro (Colocasia esculenta Schott) in Korea

  • Kim, Min-Kyu;Kwon, Soon-Bae;Yoon, Ju-Yeon;Ryu, Ki-Hyun;Heo, Su-Jeong;Hong, Jeong-Ki;Kim, Kyung-Hee;Park, Jang-Kyung
    • The Plant Pathology Journal
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    • 제20권2호
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    • pp.135-141
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    • 2004
  • A filamentous virus was isolated from taro (Colocasia esculenta Schott) showing mosaic and chlorotic feather-ing symptoms in Chuncheon, Gangwon province in 2002. Based on ELISA, its appearance in electron microscope, serological relationships, and RT-PCR using specific primer and nucleotide sequence analysis of the CP gene, the isolated virus was identified as Dasheen mosaic virus (DsMV) and designated as Korean isolated (DsMV-Kr). DsMV was not serologically related to Zantedeschia mosaic virus (ZaMV), which has been reported to infect an Araceae plants. Since the coat protein revealed electrophoretic heterogeneity, about 42 kDa, 39 kDa and 31 kDa by SDS-PAGE, an improved purification method was established for the production of antisera against DsMV-Kr. The purification method used in this study may be effectively applied to the purification of other filamentous viruses.

Efficient Method for the Rapid Purification of Nosema ceranae Spores

  • Kim, Dong-Jun;Yun, Hwi-Geon;Kim, In-Hui;Gwak, Won-Seok;Woo, Soo-Dong
    • Mycobiology
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    • 제45권3호
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    • pp.204-208
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    • 2017
  • Nosema ceranae is an obligate intracellular fungal parasite that causes mortality in honey bees and enhances the susceptibility of honey bees to other pathogens. Efficient purification of Nosema spores from the midgut of infected honey bees is very important because Nosema is non-culturable and only seasonably available. To achieve a higher yield of spores from honey bees, in this study, we considered that the initial release of spores from the midgut tissues was the most critical step. The use of 2 mm beads along with enzymatic treatment with collagenase and trypsin enhanced the homogenization of tissues and the yield of released spores by approximately 2.95 times compared with the use of common 3 mm beads alone. The optimal time for the enzyme treatment was determined to be 1 hr as measured by the yield and viability of the spores. A one-step filtration using a filter paper with an $8-11{\mu}m$ pore size was sufficient for removing cell debris. This method may be useful to purify not only N. ceranae spores but also other Nosema spp. spores.

챔버 내측에 스프링형상을 갖는 유수형 자외선 살균장치 시뮬레이션 (Simulation for the Flowing Water Purification with Spring Shape Inside Chamber)

  • 정병균;정병호;이진종;정병수
    • 전기학회논문지P
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    • 제59권4호
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    • pp.411-416
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    • 2010
  • Interest in application of ultraviolet light technology for primary disinfection of potable water in drinking water treatment plants has increased significantly in recent years. The efficacy of disinfection processes in water purification systems is governed by several key factors, including reactor hydraulics, disinfectant chemistry, and microbial inactivation kinetics. The objective of this work was to develop a computational fluid dynamics(CFD) model to predict velocity fields, mass transport, chlorine decay, and microbial inactivation in a continuous flow reactor. In this paper, It describe the how to design optimal UV disinfection device for ground water, BWT and rainwater. Spring shape instrument silver coated located in inner side of disinfection chamber. It make lead the active flowing movement target water and maximize disinfection performance. To search the optimal design method, it was performed computer simulation with 3D-CFD discrete ordinates model and manufactured prototype. Using proposed design method, performed simulation and proved satisfied performance.

The rapid synthetic strategy of [11C]PIB via disposable column cartridge purification

  • Jihye Lee;Yansheng Li;Sang-Yoon Lee;Tatsuo Ido
    • 대한방사성의약품학회지
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    • 제6권2호
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    • pp.69-74
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    • 2020
  • PIB is the first amyloid plaque PET image tracer reported for the first time in 2003, and is considered to be the best and is still being utilized due to its very high uptake and kinetic properties. Initially, it was synthesized by radioisotope labeling using a precursor containing a methoxy methyl protection group, but now it is synthesized using a 6-OH precursor that can be easily synthesized in one step using [11C]methyl triflate. Carbon-11 has several limitations in clinical studies using PET because its half-life is as short as 20 minutes. In this study, in order to overcome the difficulty of this half-life, a rapid method using Sep-Pak was adopted instead of HPLC purification to significantly reduce the burden of the purification process and attempted synthesis. As a result, the synthesis time was shortened by more than 50%, and the yield of the final compound was higher than the previous result and showed relatively high specific radioactivity, confirming that it is a strategic method with high applicability for various precursors having primary amines.

축산폐수의 흡착산화 처리 (Treatment of Animal Wastewater with Absorbent Oxidation)

  • 오인환;박정현;이명규;전병태;김형화
    • 생물환경조절학회지
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    • 제3권2호
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    • pp.136-144
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    • 1994
  • A long- time aeration method was developed for purification of animal wastewater. Under repeated aereations of 4 hours on and 4 hours off, the higher removal rates were obtained which were in average of 99%, 96%, 92% and 50% for BOD, SS, total nitrogen and phosporous, respectively. In detail, the measured BOD concentrations of the influent and effluent were 2,700ppm and 40ppm while the SS concentrations in the primary chamber and of the effluent were about 3,000 and 110 ppm, respectively. Zeolite and activated carbon, applied for removing the nitrogen and phosphorous, showed a good absorption, especially zeolite for NH$_4$-N and activated carbon for NO$_3$-N and PO$_4$-P. The treatment cost per head by this method amounts to 1,923 won and it comes to 1.6% in the whole production cost. Therefore, this method is economically available with the half cost of the conventional activated sludge process.

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다공성 세라믹과 제올라이트를 활용한 수질정화미디어블럭의 효과 연구 (A Study on Water Purification Effect of Media Block Using Porous Ceramics and Zeolite)

  • 전성률;구본학
    • 한국환경복원기술학회지
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    • 제20권5호
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    • pp.59-66
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    • 2017
  • Preeminent water treatment plans are essential to preserve the water quality of aquatic biotopes. Previous studies have not been sufficient to provide cost-effective maintenance method since they focused only on the purification of deteriorated water that requires a continuous supply of clean water. This study proposes an economical method of water quality maintenance using water treatment media block constructed vertically using porous ceramics, zeolite, and river pebble. The water treatment media block does not require a separate purification area because it functions as a purifier within the ecological pond which can maximize the biotope area. To evaluate the performance of the water treatment media block, we longitudinally tracked the change of water quality indicators (pH, TDS, COND, DO, T-P, T-N, COD) suggested by Water Environment Standards, Ministry of Environment, Republic of Korea. We compared the water quality of one control (A: general ecological pond composition method of the laminated structure) and two experimental groups (B: a combination of aquatic plants and a water treatment media block, C: a water treatment media block only). As a result, we confirmed that the water treatment media block is an efficient and economical method to maintaining the water quality of the ecological pond for a long time. The water treatment media block will be a great help in providing a better aquatic biotope space for aquatic insects and fishes living in clear water.

가잠에서 분리된 새로운 미포자충 K79의 병리학적 연구 I. 미포자충 포자의 정제 및 식별 (Pathological Studies on the New Microsporidia K79 Isolated from the Silkworm, Bombyx mori L. I. Purification and Serological Discrimination of Microsporidian Spores.)

  • 윤재수;임종성
    • 한국잠사곤충학회지
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    • 제26권2호
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    • pp.1-6
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    • 1984
  • 미포자충 포자의 정제법을 개발하고 포자표면의 혈청학적 반응을 구명하기 위하여 수행한 시험결과를 요약하면 다음과 같다. 1. 미포자충 포자의 분리정제는 예비원심-percoll 평형밀도원심-세척의 3단계법이 우수하였고, 분리정제된 포자는 극사추출에 나쁜 영향도 받지 않았다. 2. 포자의 형태는 K79 포자가 장타원형(장단경비 2.76)이고 N. bombycis는 단타원형(장단경비 2.07)으로 양포자가 상이하였다. 3. 형광항체 간접법에 의하여 항 N. bombycis 포자혈청에 대하여는 N. bomvycis 포자만이 강한 형광을 발현하였으나 항 K79 포자의 혈청에 대하여는 10배희석구에서는 양포자 모두 강하게, 20배 이상 희석구에서는 약하게 형광을 발현하였고 K79 포자의 특이항체가 포자외각에 인정되었다.

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