• Title/Summary/Keyword: Production and Purification

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Purification of Recombinant Human Alpha-2a Interferon Without Using Monoclonal Antibodies

  • Kim, Dong Chung;Jin Jung
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.916-920
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    • 2002
  • This report describes a high-level expression of human alpha-2a interferon ($IFN{\alpha}-2a$) in Escherichia coli and its pilot scale purification by using a monoclonal antibody-independent chromatographic procedure that is based on anion-exchange, cation-exchange, hydrophobic interaction, and gel filtration. The recombinant E. coli produced much more $IFN{\alpha}-2a$ in a soluble form, when cultivated at low temperatures than at high-temperature fermentation. However, if the bacterial growth was taken into consideration, fermentation at $30^{\circ}C$ seemed optimal for the interferon production. By using our new protocol, we recovered approximately 160 mg of $IFN{\alpha}-2a$ with a specific activity of $3.59{\times}10^8$ IU/mg from 201 of the broth. The gel permeation chromatographic and SDS-PAGE indicated that the interferon preparation was purified to homogeneity and was of the correctly folded fast-migrating monomer.

Development of Separation Process for Active Ingradient from Waste Biomass (폐 바이오매스로부터 생리활성물질의 분리공정 개발)

  • Sung, Ju-Li;Kim, Seong-Mun;Kim, Jin-Hyun
    • Proceedings of the Korean Institute of Resources Recycling Conference
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    • 2003.10a
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    • pp.196-200
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    • 2003
  • A novel prepurification method was developed aiming at increasing yield and purity, also reducing solvent usage for purification of paclitaxel. The use of a micelle and precipitation in the prepurification process allows for rapid separation of paclitaxel from interfering compounds and dramatically reduces solvent usage compared to alternative methodologies. The prepurification process serves to minimize the size and complexity of the HPLC operations for paclitaxel purification. The process is readily scalable to a pilot plant and eventually to a production environment where multikilogram quantities of material are expected to be produced. As much as possible, the process has been optimized to minimize solvent usage, complexity, and operating costs.

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Methods for High Quality Purification of Alginate and Fucoidan from Marine Brown Algae (해조류로부터 푸코이단/알지네이트의 고순도 분리정제)

  • Kim, Jong-Ki;Yang, Jae-Ho
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.4
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    • pp.245-249
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    • 2007
  • High quality of purified alginate and fucoidan is required for the medical uses to prevent the unexpected side effects from the contaminated endotoxin present in the materials. We attempted to establish an efficient and fast mass scale production method for the highly purified poly-G alginate from the sea weeds. In addition, lab scale method was established to obtain the fraction of higher purity from the commercially available fucoidan source (about 85% purity).

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Purification of Carp Serum Metallothionein Induced in Carp with Cadmium and Production of Polyclonal Antibody (카드뮴 유도 잉어 혈액으로부터의 Metallothionein 정제 및 이에 대한 다클론 항체 생산)

  • Ryu, Hyung-Seok;Kang, Ho-Joon;Kim, Seul Ki;Kim, Namsoo;Kim, Woo-Yeon
    • Food Engineering Progress
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    • v.13 no.4
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    • pp.314-319
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    • 2009
  • Metallothioneins (MTs) are low molecular weight, cysteine-rich, and heavy metal binding proteins, which could be induced with heavy metals such as Cd, Hg, Zn and Cu in liver, kidney, and in cultured cells. By using ion exchange chromatography on DE-52, MT was purified from the serum of carp induced with cadmium in order to produce antibody against MT. Polyclonal antibody produced against purified carp MT reacted well with MT in the serum of carp induced with cadmium, whereas control serum did not. This may indicate that the polyclonal antibody against the carp MT could be used for the preparation of biosensors to detect MT in fishes like carp.

Purification of Phytase from Aspergillus ficuum and Production of Anti-phytase Antibody (Aspergillus ficuum의 Phytase의 정제와 Anti-phytase 항체생산)

  • Kim, Keun
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.299-303
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    • 1999
  • Phytase(myo-inositol-hexakis phosphate 3-phosphohydrolase, E C 3.1.3.8) sequentially hydrolyzes phytate to myo-inositol and inorganic phosphate. Phytase of Aspergillus ficuum was purified to homogeneity using ultrafiltration, cation exchange column and anion exchange column. It's molecular weight is estimated as around 90,000 by SDS-PAGE. Antibody against the phytase was produced by immunizing mice with the purified phytase. The titer of the antibody was determined to be 1/25,000.

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Simple Purification of BA-RGD Protein Based on CaCl2/EDTA Treatment and Inclusion Body Washing (CaCl2/EDTA 및 비이온성 계면활성제 활용 Inclusion Body 정제법을 이용한 BA-RGD 단백질의 생산)

  • Song, Wooho;Byun, Chang Woo;Yoon, Minho;Eom, Ji Hoon;Choi, Yoo Seong
    • KSBB Journal
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    • v.30 no.6
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    • pp.291-295
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    • 2015
  • The limited productivity of natural shell matrix proteins has hampered the investigation of their biochemical properties and practical applications, although biominerals in nature obtained by organic-inorganic assemblies have attractive mechanical and biological properties. Here, we prepared a vector for the expression of a fusion protein of a shell matrix protein from Pinctada fucata (named as GRP_BA) with the GRGDSP residue. The fusion protein of BA-RGD was simply produced in E. coli and purified through sequential steps including the treatment with $CaCl_2$ and EDTA solution for cell membrane washing, mechanical cell disruption and the application of non-ionic surfactant of Triton X-100 for BA-RGD inclusion body washing. The production yield was approximately 60 mg/L, any other protein band was not observed in SDS-PAGE and it was estimated that above 97% endotoxin was removed compared to the endotoxin level of whole cell. This study showed this simple and easy purification approach could be applied to the purification of BA-RGD fusion protein. It is expected that the protein could be utilized for the preparation of biominerals in practical aspects.

A Study on the Design Factors of the Cylinder-Shaped Filter for Improving Reservoir Water Quality (호소수질 개선을 위한 원통형 여과장치의 설계요소 연구)

  • Lee, Sun-Ho;Kim, Seung-Hyun
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.8
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    • pp.655-662
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    • 2009
  • In order to obtain knowledge on the design and operation of practical?scale Cylinder-Shaped Filters, pilot experiments were conducted to observe the effects of stepwise augmentation of production rate on water quality and clogging. A production rate increase from 0.8 L/min to 2.4 L/min did not appear to affect the removal efficiencies of BOD(Biochemical Oxygen Demand), SS(Suspended Solids), and turbidity, as the values were maintained around 80%, over 95%, and over 90% respectively;however, COD(Chemical Oxygen Demand) removal decreased from 44% to 29%. In addition, results indicated an inverse relationship between production rate and detention time required to remove unit contaminant concentration, the observed detention time in the filter to remove 1 mg-COD/L being 83 minutes for the production rate of 0.8 L/min and 45 minutes for the production rate of 2.4 L/min, suggesting that a relatively higher production rate is likely to be more advantageous in the purification of reservoir water when compared to a lower production rate. Clogging was observed to originate from the surface and advance to the center of the filter, and although clogging seemed to increase as the production rate increased, this did not cause any difficulties in normal functioning of the filter for more than 2 years of operation, suggesting that this filter system can be used effectively in the purification of reservoir water.

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

  • Cho, Min;Ryu, Yeon-Woo
    • KSBB Journal
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    • v.14 no.2
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    • pp.248-254
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    • 1999
  • Adverse reactions after injection of diphtheria vaccine are induced by impurities present in crude toxoids that cannot be removed completely by purification of toxoids after formalization. To increase toxoid purity, toxin purification was tried before formalization. Crude toxin was purified with ultrafiltration and ion-exchange chromatography. Purified toxin purity was improved 2.9 times higher than crude toxin, and purity was 2,560 Lf/mg PN. Purified toxin was detoxified with formalin and lysine, and potency test were performed. Toxoid, prepared from toxin treated with formalin and lysine, did not show reversion to toxin and purity was higher than the toxoid purified after formalization. Therefore, we concluded that the use of toxoid vaccine prepared from toxin purified is a useful method of minimize adverse reaction after injection of diphtheria vaccine.

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Impacts of Mt. Bongeui on Atmospheric Purification in Chuncheon

  • Piao, Zhi Hui;Jo, Hyun-Kil
    • Journal of Forest and Environmental Science
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    • v.34 no.2
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    • pp.165-168
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    • 2018
  • Mt. Bongeui is a neighborhood park of 66.4 ha as a major natural landscape resource located in the center of Chuncheon City. This study quantified the reduction of carbon, $SO_2$, $NO_2$, $O_3$, and $PM_{2.5}$, and the production of $O_2$ to explore the role for Mt. Bongeui to contribute to annual atmospheric purification. The main forest types and age classes of the study site included age-class III broadleaved forest at 35.8%, age-class VI coniferous forest at 17.2%, and age-class IV broadleaved forest at 15.7%. The annual atmospheric purification effect per unit area was as follows: 7.6 t carbon/ha/yr, 16.6 kg $SO_2/ha/yr$, 40.4 kg $NO_2/ha/yr$, 41.5 kg $O_3/ha/yr$, 53.7 kg $PM_{2.5}/ha/yr$, and 20.2 t $O_2/ha/yr$. The economic value of these effects was equivalent to about 12.9 million KRW/ha/yr. The study site annually offset carbon emissions of about 300 citizens, $SO_2$ emissions of 220 citizens, and $NO_2$ emissions of 92 citizens in Chuncheon. It also played an important role in annually producing 1.5% of the amount of $O_2$ necessary for the respiration of Chuncheon's total population. This study pioneers in comprehensively quantifying the atmospheric purification effect and could be useful in guiding the planning and management to improve the effect.

Purification and Characterization of Paclitaxel from Plant Cell Cultures of Taxus chinensis in Large-Scale Process (식물세포 Taxus chinensis 배양으로부터의 Paclitaxel 대량 정제 및 특성)

  • 김진현;기은숙;민범찬;최형균;홍승서;이현수
    • KSBB Journal
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    • v.15 no.5
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    • pp.537-540
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    • 2000
  • In developing a HPLC purification process, it was hoped that a single chromatographic system would be sufficient to abtain pure paclitaxel in high yield. However, no such system was found, due in part to the complex taxoid profile of crude paclitaxel and to the rigorous nature of the product specification. A two step HPLC purification was adopted using reverse-phase separation on C(sub)18 as a first step, and normal-phase separation on silica as the final polishing step. Impurity profiles were established and maintained for paclitaxel, which identified and quantified each impurity observed in purified paclitaxel from these two steps, all impurities at or above 0.1% were identified. Results provide information for improving the quality control of paclitaxel production.

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