• Title/Summary/Keyword: Solid-phase adsorption

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Solid-Phase Refolding of Inclusion Body Protein in Packed Bed Adsorption and Expanded Bed Adsorption Chromatography (Packed Bed Adsorption과 Expanded Bed Adsorption 크로마토그래피를 이용한 내포체 단백질의 고체상 재접힘)

  • 최원찬;김민영;서창우;이은규
    • KSBB Journal
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    • v.18 no.6
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    • pp.500-505
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    • 2003
  • ‘LK (lipoprotein kringle) 68’is a polypeptide of a modified ansiostatin consisting of three kringle structures that might be clinically useful as a potential cancer therapeutics. It can be produced by overexpressing it as inclusion body in recombinant E. coli. In this study, solid-phase refolding processes using packed bed adsorption (PBA) and expanded bed adsorption (EBA) column were carried out to compare their refolding yields with that of the conventional, solution-phase refolding process, For the solution-phase and the PBA-mediated processes employing Q-Sepharose, washed inclusion body was used as the starting material, whereas both washed inclusion body and E. coli homogenate were used for the EBA-mediated process employing streamline DEAE. On the final recovery LK68 per unit mass of wet cell basis, the EBA- and PBA-mediated processes showed about 2.7- and 1.5-fold higher yields, respectively, than the solution-phase refolding method. The solid-phase refolded LK68 demonstrated the same Iysine binding bioactivity and the retention time in the RP-and SEC-HPLC as those of the native protein.

Optimization of Headspace Sampling Using Solid Phase Microextraction For Volatile Organic Acids in Different Tobacco Types

  • Lee, Jang-Mi;Lee, Jeong-Min;Son, Seong-Ae;Kwon, Young-Ju;Jang, Gi-Chui;Kim, Young-Ho
    • Journal of the Korean Society of Tobacco Science
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    • v.35 no.1
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    • pp.7-12
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    • 2013
  • A Solid-phase micro extraction(SPME) was evaluated as a tool for headspace sampling of tobacco samples. Several experimental parameters (sampling temperature, pH, and type of SPME fibers) were optimized to improve sampling efficiency in two aspects ; maximum adsorption and selective adsorption of volatile organic acids onto SPME fibers. Among four types of SPME fibers such as PDMS(Polydimethylsiloxane), PA(Polyacrylate), Car/PDMS (Carboxen/Polydimethylsiioxane) and PDMS/DVB(Polydimethylsiioxane/Divinylbenzene) which were investigated to determine the selectivity and adsorption efficiency. A variety of tobacco samples such as flue cured, burley and oriental were used in this study. The effect of these parameters was often dominated by the physical and chemical nature (volatility, polarity) of target compounds. This method allowed us to make important improvements in selectivity and sensitivity. The Car/PDMS fiber was shown to be the most efficient at extracting the 10 selected volatile organic acids. The parameters were optimized: $80^{\circ}C$ adsorption temperature, 30 min of adsorption time, $240^{\circ}C$ desorption temperature, 1 min of adsorption time.

Determination of Siloxanes in Biogas by Solid-phase Adsorption on Activated Carbon

  • Kim, Nack-Joo;Chun, Seung-Kyu;Cha, Daniel K.;Kim, Cheal
    • Bulletin of the Korean Chemical Society
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    • v.34 no.8
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    • pp.2353-2357
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    • 2013
  • The objective of this study was to develop a simple, less time-consuming and accurate sampling technique based on solid-phase sorption with activated carbon as the sorbents. The results from solid-phase sorption techniques were compared to that from a conventional solvent impinger-based technique to confirm the efficacy of the proposed method. The laboratory results indicated that the solid-phase sorption method was suitable for the determination of siloxanes as the measured concentrations were similar to that from a solvent impinge method. The data from solid-phase sorption method showed excellent recovery and reproducibility while the sampling was less labor intensive and less time consuming than the solvent impinge method. Following the laboratory tests, the solid-phase sorption technique was successfully applied to sampling biogas from a field site. This study shows that the activated carbon-based solid-phase sorption can be a reliable and less time-consuming option for the sampling and collection of siloxanes under various different landfill conditions.

In Vitro Refolding of Inclusion Body Proteins Directly from E. coli Cell Homogenate in Expanded Bed Adsorption Chromatography (Expanded Bed Adsorption 크로마토그래피를 사용하여 재조합 E. coli 세포 파쇄액으로부터 내포체 단백질을 직접 재접힘하는 공정)

  • 조태훈;서창우;이은규
    • KSBB Journal
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    • v.16 no.2
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    • pp.146-152
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    • 2001
  • To avoid the intrinsic problem of aggregation associated with the traditional solution-phase refolding process, we propose a solid-phase refolding method integrated with expanded bed adsorption chromatography. The model protein used was a fusion protein of recombinant human growth hormone and a glutathione S transferase fragment. It was demonstrated that the EBA-mediated refolding technique could simultaneously remove cellular debris and directly renature the fusion protein inclusion bodies in the cell homogenate with much higher yields and less agregation. To demonstrate the applicability of the method, we successfully tested the three representative types of starting materials, i. e., rhGH monomer, washed inclusion bodies, and the E. coli homogenate. This direct and simplified refolding process could also reduce the number of renaturation steps required and allow refolding at a higher concentration, at approximately 2 mg fusion protein per ml of resin. To the best of our knowledge, it is the first approach that has combined the solid-phase refolding method with expanded bed chromatography.

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Stepwise Adsorption in Gas-Solid Adsorption System and Phase Transition in Adsorbed Phase

  • Lee Song Hee;Lee Jo Woong;Pak Hyungsuk;Chang Seihun
    • Bulletin of the Korean Chemical Society
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    • v.3 no.2
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    • pp.37-44
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    • 1982
  • In this work we have studied the multilayer stepwise adsorption of gases on solid adsorbents based on the previously developed theory. It is shown that stepwise adsorption isotherms emerge from our theory if an ad hoc adsorption regarding the degree of occupation for each successive layer is abolished and the effect of lateral intermolecular interactions among adsorbate molecules is included. In addition to these the effect of vertical interactions has also been taken into consideration. It seems that the vertical interaction plays a role in deciding the shape and the position of steps in resulting isotherms. It is evident from this research that it is the lateral interaction that is responsible for stepwise adsorption as long as the adsorbent surface is uniform and temperature is sufficiently low.

Characterization of Adsorbents for the Solid-Phase Extraction of Trace Ibuprofen from Biological Samples (생체시료로부터 미량 이부푸로펜의 고상추출에 사용될 흡착제들의 특성)

  • Kim, Kyoung-Rae;Shim, Weon-Hee
    • YAKHAK HOEJI
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    • v.36 no.6
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    • pp.563-569
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    • 1992
  • The sorption and desorption properties of four different solid adsorbents were evaluated for the trace enrichment of ibuprofen from biological samples. Chromosorb 107 gave the highest dynamic adsorption coefficient. Among the organic solvents examined, acetone gave the highest desorption coefficient. Using the Chromosorb 107 column, the optimum elution volume of the eluting solvent was evaluated from the breakthrough curve of ibuprofen. The usefulness of Chromosorb 107 as the solid adsorbent and acetone as the eluting solvent was examined for the solid-phase extraction of ibuprofen from serum in the concentration range of $20{\sim}40\;{\mu}g/ml$.

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A Study on the Analytical Methods Using Solid-phase Extraction for the Determination of Phenols in Water

  • Jeon, Chi-Wan;Lee, Jung-Hwa;Lee, Sang-Hak
    • Proceedings of the Korean Environmental Sciences Society Conference
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    • 2003.11a
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    • pp.135-138
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    • 2003
  • Based on solid phase extraction, gas chromatography and mass spectrometry procedure for determining phenol and its derivatives in natural water was presented. In solid phase extraction, three types of techniques using solid phase adsorption material were treated with acid and salt, and converted second portion of acetyl derivatives. Under the these condition, extraction efficiency and detection ability dependent on extraction methods were discussed. Obtained results using optimized solid phase extraction techniques showed more convenience, simplifier and lower cost than the conventional analytical methods with holding wide dynamic range and lower detection limits.

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Analysis of Residual Solvents in Food Packaging Materials Using Solid Phase Microextraction Method (Solid Phase Microextraction법을 이용한 식품포장재 중의 잔류용제 분석)

  • 서택교;박상현;이윤수;김정한;권익부
    • Journal of Food Hygiene and Safety
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    • v.14 no.1
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    • pp.76-83
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    • 1999
  • Solid phase microextraction (SPME) was used for the determination of 6 standard solvents (methanol, isopropanol, methyl ethyl ketone, ethyl acetate, cyclohexane, toluene) in food packaging materials. SPME method is a solvent-free sample preparation technique in which a fused silica fiber coated with polymeric organic liquid is introduced into the headspace above the sample. SPME method using fiber coated polydimethylisiloxane (PDMS) was compared with static headspace (SHS) method used as a reference. It was found that the optimal adsorption condition using PDMS-SPME method was 2$0^{\circ}C$ for 15 minutes for the standard solvents. Detection limits, linearity, reproducibility and recovery of both SHS and PDMS-SPME methods have been determined using 6 standard solvents. Both methods were characterized by high reproducibility and good linearity. Using SHS methods, the mean recovery of the 6 standard solvents was ranged from 75.5% to 105.8% with a mean relative standard deviation (RSD) of 0.3% to 4.8%. With PDMS-SPME method, the mean recovery of the 6 standard solvents was ranged from 86.7% to 108.3% with a mean RSD of 0.4% to 2.5%. The detection limits of both methods were the same for toluene, cyclohexane and methyl ethyl ketone; those of PDMS-SPME method were higher than those of SHS method for methanol, isopropanol and ethyl acetate. PDMS-SPME fiber shoed excellent adsorption for non-polar solvents such as toluene, while it showed relatively low adsorption for polar solvents such as methanol.

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Fabrication and Characterisation of a Novel Pellicular Adsorbent Customised for the Effectvie Fluidised Bed Adsorption of Protein Products

  • Sun, Yam;Pacek, Andrzej W.;Nienow, Alvin W.;Lyddiatt, Andrew
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.419-425
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    • 2001
  • A dense pellicular solid matrix has been fabricated by coating 4% agarose gel on to dense zironia-silica(ZS) spheres by watr-in-oil emulsification . The agarose evenly laminated the ZS bead to a depth of 30㎛, and the resultin gpellicular assembly was characterised by densities up to 2.39g/mL and a mean particle dimeter of 136 ㎛. In comparative fluidisation tests, the pellicular solid phase exhibited a two-fold greater flow velocity than commercial benchmark ad-sorbents necessary to achieve common values of bed expansion. Furthermore, the perlicular parti-cles were characterised by improved qualities of chromatographic behaviour, particularly with re-spect to a three-fold increase in the apparent effective diffusivity of lysozyme within a pellicular assembly modified with Cibacron Blue 3GA. The properties of rapid protein adsorption/desorp-tion were attributed to the physical design and pellicular deployment of the reactive surface in the solid phase. When combined with enhanced feedstock throughput, such practical advantages recommend the pellicular assembly as a base matrix for the selective recovery of protein products from complex, particulate feedstocks(whole fermentation broths, cell disruptates and biological extracts).

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Synthesis and characterization of α-mangostin imprinted polymers and its application for solid phase extraction

  • Zakia, Neena;Zulfikar, Muhammad A.;Amran, Muhammad B.
    • Advances in materials Research
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    • v.9 no.4
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    • pp.251-263
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    • 2020
  • α-mangostin imprinted polymers have been synthesized by a non-covalent imprinting approach with α-mangostin as a template molecule. The α-mangostin molecularly imprinted polymers (MIPs) prepared by radical polymerization using methacrylic acid, ethlylene glycol dimethacrylate, benzoyl peroxide, and acetonitrile, as a monomer, crosslinker, initiator, and porogen, respectively. The template was removed by using methanol:acetic acid 90:10 (v/v). The physical characteristics of the polymers were investigated by Fourier Transform Infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and thermogravimetric analysis (TGA). The rebinding studies were carried out by batch methods. The results exhibited that the MIPs was able to adsorb the α-mangostin at pH 2 and the contact time of 180 min. The kinetic adsorption data of α-mangostin performed the pseudo-second order model and followed the Langmuir isotherm model with the adsorption capacity of 16.19 mg·g-1. MIPs applied as a sorbent material in solid-phase extraction, namely molecularly imprinted solid-phase extraction (MISPE) and it shows the ability for enrichment and clean-up of α-mangostin from the complex matrix in medicinal herbal product and crude extract of mangosteen (Garcinia mangostana L.) pericarp. Both samples, respectively, which were spiked with α-mangostin gives recovery more than 90% after through by MISPE in all concentration ranges.