• Title/Summary/Keyword: BSA-silica

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Chiral Separation of Tryptophan Enantiomers by Liquid Chromatography with BSA-Silica Stationary Phase

  • Kim Kwonil;Lee Kisay
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.17-22
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    • 2000
  • The separation of tryptophan enantiomers was carried out with medium-pressure liquid chromatography using BSA (bovine serum albumin)-bonded silica as a chiral stationary phase. The influence of various experimental factors such as pH and ionic strength of mobile phase, separation temperature, and the presence of organic additives on the resolution was studied. In order to expand this system to preparative scale, the loadability of sample and the stability of stationary phase for repeated use were also examined. The separation of tryptophan enantiomers was successful with this system. The data indicated that a higher separation factor (a) was obtained at a higher pH and lower temperature and ionic strength in mobile phase. Addition of organic additives (acetonitrile and 2-propanol) in mobile phase contributed to reduce the retention time of L-tryptophan. About $30\%$ of the separation factor was reduced after 80 days of repeated use.

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Affinity Filtration Chromatography of Proteins by Chitosan and Chitin Membranes: 2. Separation of BSA and Lysozyme (키토산 및 키틴 막에 의한 단백질의 친화 여과 크로마토그래피: 2. BSA 및 Lysozyme의 분리)

  • Youm, Kyung-Ho;Yuk, Yeong-Jae
    • Membrane Journal
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    • v.19 no.2
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    • pp.113-121
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    • 2009
  • Porous affinity chitosan and chitin membranes with good mechanical strength and high protein binding capacity were prepared by using silica particles as porogen. The maximum binding capacity of affinity chitosan membrane for BSA protein is 21.8mg/mL, and that of affinity chitin membrane for lysozyme enzyme is 26.1mg/mL. Chromatographic separations of BSA and lysozyme proteins using the porous affinity chitosan and chitin membranes were performed with change of the flow rate, loading amount and concentration of protein loading solutions. Protein eluted amount and binding yield were calculated from the filtration chromatograms consisted of loading/washing/elution sequences. Protein binding amount and yield were increased with decreasing of flow rate, increasing of loading amount and concentration of protein loading solutions. Those results suggest that the porous chitosan and chitin membranes prepared by using silica particles as porogen are suitable in affinity filtration chromatography for large scale separation of proteins.

Preparation for Protein Separation of an Ion-Exchange Polymeric Stationary Phase Presenting Amino Acid and Amine Units Through Surface Graft Polymerization

  • Choi Seong-Ho;Lee Kwang-Pill;Shin Chang-Ho
    • Macromolecular Research
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    • v.13 no.1
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    • pp.39-44
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    • 2005
  • Ion-exchange polymeric stationary phases presenting amino acid and amino groups were prepared by the surface grafting of glycidyl methacrylate onto a silica gel surface and subsequent amination. Three kinds of amino acids-L-arginine (Arg), D-lysine (Lys), and D-histine (His)-were used in this study. An ion-exchange polymeric stationary phase presenting ethylene diamine (EDA) was also prepared by surface graft polymerization. Separation of the model proteins bovine serum albumin (BSA), chick egg albumin (CEA), and hemoglobin (Hb) was performed using the amino acid- and amine-derived columns. In separating the CEA/BSA mixture, the resolution time of BSA was longer than that of CEA when using the EDA column, whereas the resolution time of BSA was shorter than that of CEA when using the Arg, Lys, and His columns. In the separation of the Hb/BSA mixture, the resolution time of BSA was longer than that of Hb in the EDA column, whereas the resolution time of BSA was shorter than that of Hb in the amino acid columns (D-Lys, L-Arg, and D-His).

A Simple Mehtod for Conjugation Between Ginsenoside Rf and Bovine Serum Albumin

  • Yoon So-Rah;Nah Jin-Ju;Choi Seok;Son Chang-Ho;Shin Dong-Ho;Ko Sung-Ryong;Kim Seok-Chang;Nam Ki-Yeul;Nah Seung-Yeol
    • Journal of Ginseng Research
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    • v.23 no.1 s.53
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    • pp.55-59
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    • 1999
  • For the first time we have developed a simple method for conjugation between ginsenoside Rf (Rf) and bovine serum albumin (BSA) or ovalbumin (OVA) by a periodate oxidation method. The Rf-BSA conjugate is confirmed by ultraviolate (UV) scanning, thin layer chromatography (TLC), and SDS-gel electrophoresis. In UV scanning Rf showed three small peaks approximately at 230, 265 and 280 nm. BSA showed a peak at 280 nm. The Rf-BSA conjugate showed right shifted three peaks that BSA alone did not show. In TLC analysis, the Rf-BSA conjugate did not show mobility in silica gel but showed a slight stream of trace. In SDS-gel electrophoresis, Rf-BSA conjugate show a slight less mobility than BSA alone. Rf-OVA conjugate also showed similar patterns with Rf-BSA conjugate. These results demonstrated that periodate oxidation method could be used to produce a stable Rf-BSA or Rf-OV A conjugate and also could be applied for other ginsenoside(s).

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Effect of the Crude Polysaccharides Fraction from Eleutherococcus senticosus as a Immunoadjuvant to Soluble Antigens (BSA and OVA) (오가피 조다당의 단백질 항원 (BSA and OVA)에 대한 면역증강효과)

  • 황수현;하은숙;유광원;신광순;이상훈;이재권;이경호;윤택준;박우문
    • YAKHAK HOEJI
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    • v.47 no.3
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    • pp.167-175
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    • 2003
  • Eleutherococcus senticosus is a typical oriental folk medicinal herb. It has been used clinically as a anti-rheumatic disease, anti-stress, ischemic heart disease and gastric ulcer. In the present study, we examined the adjuvant activity of the crude polysaccharides fraction from Eleutherococcus senticosus, EN-3, on the induction of humoral and cellular immune responses against bovine serum albumin (BSA) or ovalbumin (OVA). The thioglycollate-induced macrophages and silica-induced dendritic-like cells cultured with BSA and EN-3 synergistically increased the production of TNF-$\alpha$ and IL-12. When mice were subcutaneously immunized with BSA + EN-3, the antibody titer against BSA was showed significantly higher than those immunized with BSA alone. In addition, when mice were immunized with OVA + FIA + EN-3, the antibody titer was showed similar patterns with the FCA. The assay for determining subisotype of antibody revealed that EN-3 augmented OVA-specific antibody titer of IgG1 and IgG2b. The culture supernatant obtained from splenocytes of mice treated with OVA + FIA + EN-3 also showed a higher level of both OVA-specific Th1-type (IL-2, IFN-${\gamma}$ and GM-CSF) and Th2-type cytokine (IL-4, IL-6 and IL-10). In vitro analysis of T cell proliferation to OVA on 8 weeks, the splenocytes of mice treated with OVA + EN-3 showed a significantly higher proliferating activity than those treated with OVA alone. These results suggest that EN-3 may possess adjuvant activities to potentially to enhance humoral as well as cellular immune response.

Affinity Filtration Chromatography of Proteins by Chitosan and Chitin Membranes: 1. Preparation and Characterization of Porous Affinity Membranes (키토산 및 키틴 막에 의한 단백질의 친화 여과 크로마토그래피: 1. 다공성 친화 막의 제조와 특성 평가)

  • Youm Kyung-Ho;Yuk Yeong-Jae
    • Membrane Journal
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    • v.16 no.1
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    • pp.39-50
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    • 2006
  • Porous chitosan and chitin membranes were prepared by using silica particles as porogen. Membrane preparation was achieved via the following three steps: (1) chitosan film formation by casting an chitosan solution containing silica particles, (2) preparation of porous chitosan membrane by dissolving the silica particles by immersing the film into an alkaline solution and (3) preparation of porous chitin membrane by acetylation of chitosan membrane with acetic anhydride. The optimum preparation conditions which could provide a chitosan and chitin membranes with good mechanical strength and adequate pure water flux were determined. To allow protein affinity, a reactive dye (Cibacron Blue 3GA) was immobilized on porous chitosan membrane. Binding capacities of affinity chitosan and chitin membranes for protein and enzyme were determined by the batch adsorption experiments of BSA protein and lysozyme enzyme. The maximum binding capacity of affinity chitosan membrane for BSA protein is about 22 mg/mL, and that of affinity chitin membrane for lysozyme enzyme is about 26 mg/mL. Those binding capacities are about $several{\sim}several$ tens times larger than those of chitosan and chitin-based hydrogel beads. Those results suggest that the porous chitosan and chitin membranes are suitable in affinity filtration chromatography for large scale separation of proteins.

Preparation of Polysulfone Composite Ultrafiltration Hollow Fiber Membranes Incorporating Nano-size Fumed Silica with Enhanced Antifouling Properties (나노 크기의 Fumed Silica가 함유된 Polysulfone 한외여과 중공사막 제조 및 내오염성 분석)

  • Kang, Yesol;Lim, Joohwan;Kim, In S.
    • Membrane Journal
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    • v.28 no.6
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    • pp.379-387
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    • 2018
  • This study was conducted to improve the membrane characteristics and performance by increasing hydrophilicity by adding additives to the ultrafiltration polysulfone (PSf) hollow fiber membrane. The mixed matrix membranes (MMMs) were prepared by dispersing 15 nm of fumed silica (FS) in the spinning solution at 0.1, 0.3 and 0.5 wt%. SEM analysis was carried out to confirm the cross-section and surface condition. It was confirmed that mean pore radius of the hollow fiber increased by 4 nm as FS was added. In addition, contact angle measurement was carried out for the hydrophilicity analysis of hollow fiber membranes, and it was confirmed that the hydrophilicity of MMMs were increased by adding of FS. In the case of water permeability, the membrane including FS showed 91~96 LMH and showed 5~11% more increase than PSf membrane. In the antifouling performance test, relative flux reduction ratios of FS mixed hollow fiber membranes were lower than that of PSf membranes, and it was confirmed that increase of hydrophilicity hinders adsorption of hydrophobic BSA on the membrane surface.

Fouling of Ion Exchange Membranes and Their Fouling Mitigation (이온교환막의 막오염 및 오염저감)

  • 문승현;이홍주
    • Membrane Journal
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    • v.12 no.2
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    • pp.55-66
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    • 2002
  • fouling phenomena of ion exchange membranes were reviewed for improved design and operation of electodialysis. The membrane fouling index for electrodialysis (EDMFI) was defined for the quantitative analysis of fouling potential as an analogy to the pressure-driven membrane process. fouling phenomena were compared in the electrodialysis experiments with inorganic foulant (silica sol) and organic foulants (humate and bovine serum albumin (BSA)), and their fouling potentials were analyzed using the fouling index. The comparison showed that the EDMFI could be used as a quantitative measure of the fouling tendency in electrodialysis processes. As a novel fouling mitigation method, square wave power was reported to be effective in electrodialysis with organic foulants. The square wave powers having the pulsed electric field enabled to reduce the membrane fouling significantly at an optimal frequency.

Stability of PS Opals in Supercritical Carbon Dioxide and Synthesis of Silica Inverse Opals

  • Yu, Hye-Min;Kim, Ah-Ram;Moon, Jun-Hyuk;Lim, Jong-Sung;Choi, Kyu-Yong
    • Bulletin of the Korean Chemical Society
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    • v.32 no.7
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    • pp.2178-2182
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    • 2011
  • Recently, the synthesis of ordered macroporous materials has received much attention due to its potential use as photonic band gap materials.$^1$ In this study, we have used the three-dimensional (3D) latex array template impregnated with benzenesulfonic acid (BSA), which is capable of catalyzing the reaction using tetraethyl orthosilicate (TEOS) as a precursor and distilled water. The polystyrene (PS) templates were reacted with TEOS in $scCO_2$ at 40 $^{\circ}C$ and at 80 bar. In the reactor, TEOS was filtrated into the PS particle lattice. After the reaction, porous silica materials were obtained by calcinations of the template. The stability test of the PS template in pure $CO_2$ was conducted before the main experiment. Scanning electron microscopy (SEM) images showed that the reaction in $scCO_2$ takes place only on the particle surface. This new method using $scCO_2$ has advantages over conventional sol-gel processes in its capability to control the fluid properties such as viscosity and interfacial tension. It has been found that the reaction in $scCO_2$ occurs only on the particle surface, making the proposed technique as more rapid and sustainable method of synthesizing inverse opal materials than conventional coating processes in the liquid phase and in the vapor phase.

Highly Sensitive Detection of Pathogenic Bacteria Using PDMS Micro Chip Containing Glass Bead (유리비드를 포함한 PDMS 마이크로칩을 이용한 고감도 감염성 병원균 측정에 관한 연구)

  • Won, Ji-Yeong;Min, Jun-Hong
    • KSBB Journal
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    • v.24 no.5
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    • pp.432-438
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    • 2009
  • Here, we demonstrated simple nucleic acid, RNA, concentration method using polymer micro chip containing glass bead ($100\;{\mu}m$). Polymer micro chip was fabricated by PDMS ($1.5\;cm\;{\times}\;1.5\;cm$, $100\;{\mu}m$ in the height) including pillar structure ($160\;{\mu}m\;(I)\;{\times}\;80\;{\mu}m\;(w)\;{\times}\;100\;{\mu}m\;(h)$, gap size $50\;{\mu}m$) for blocking micro bead. RNA could be adsorbed on micro glass bead at low pH by hydrogen bonding whereas RNA was released at high pH by electrostatic force between silica surface and RNA. Amount of glass beads and flow rate were optimized in aspects of adsorption and desorption of RNA. Adsorption and desorption rate was measured with real time PCR. This concentrated RNA was applied to amplification micro chip in which NASBA (Nucleic Acid Sequence Based Amplification) was performed. As a result, E.coli O157 : H7 in the concentration of 10 c.f.u./10 mL was successfully detected by these serial processes (concentration and amplification) with polymer micro chips. It implies this simple concentration method using polymer micro chip can be directly applied to ultra sensitive method to measure viable bacteria and virus in clinical samples as well as environmental samples.