• Title/Summary/Keyword: affinity beads

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Selective adsorption of Ba2+ using chemically modified alginate beads with enhanced Ba2+ affinity and its application to 131Cs production

  • Kim, Jin-Hee;Lee, Seung-Kon
    • Nuclear Engineering and Technology
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    • v.54 no.8
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    • pp.3017-3026
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    • 2022
  • The 131Cs radioisotope with a short half-life time and high average radiation energy can treat the cancer effectively in prostate brachytherapy. The typical 131Cs production processes have a separation step of the cesium from 131Ba to obtain a high specific radioactivity. Herein, we suggested a novel 131Cs separation method based on the Ba2+ adsorption of alginate beads. It is necessary to reduce the affinity of alginate beads to cesium ions for a high production yield. The carboxyl group of the alginate beads was replaced by a sulfonate group to reduce the cesium affinity while reinforcing their affinity to barium ions. The modified beads exhibited superior Ba2+ adsorption performances to native beads. In the fixed-bed column tests, the saturation time and adsorption capacity could be estimated with the Yoon-Nelson model in various injection flow rates and initial concentrations. In terms of the Cs elution, the modified alginate showed better performance (i.e., an elution over 88%) than the native alginate (i.e., an elution below 10%), indicating that the functional group modification was effective in reducing the affinity to cesium ions. Therefore, the separation of cesium from the barium using the modified alginate is expected to be an additional option to produce 131Cs.

Methacrylamidohistidine in Affinity Ligands for Immobilized Metal-ion Affinity Chromatography of Human Serum Albumin

  • Odaba, Mehmet;Garipacan, Bora;Dede, Semir;Denizli, Adil
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.6 no.6
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    • pp.402-409
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    • 2001
  • Different bioligands carrying synthetic adsorbents have been reported in the literature for protein separation, We have developed a novel and new approach to obtain high protein ad-sorption capacity utilizing 2-methacrylamidohistidine(MAH) as a bioligand. MAH was synthe-sized by reacting methacrylocholride and histidine, Spherical beads with an average size of 150-200㎛ were obtained by the radical suspension polymerization of MAH and 2-hydrosyethyl-methacrylate(HEMA) conducted in an aqueous dispersion medium. p(HEMA-co-MAH) beads had a specific surface area of 17.6㎡/g . Synthesized MAH monomer was characterized by NMR. p(HEMA-co-MAH) beads were characterized by swelling test, FTIR and elemental analysis. Then Cu(II) ions were incorporated onto the beads and Cu(II) loading was found to be 0.96 mmol/g.These affinity beads with a swelling ration of 65% and containing, 1.6 mmol MAH/g were used in the adsorption/desorption of human serum albumin(HSA) from both aqueous solutions and hu-man serum. The adsorption of HSA onto p(HEM-co-MAH) was low(8.8 mg/g). Cu(II) chelation onto the beads significantly increased the HSA adsorption (56.3 mg/g). The maximum HSA ad-sorption ws observed at pH 8.0 Higher HSA adsorption was observed from human plasma(94.6 mgHSA/g) Adsorption of other serum proteins were obtained as 3.7 mg/g for fibrinogen and 8.5mg/g for γ-globulin. The total protein adsorption was determined as 107.1mg/g. Desorption of HSA was obtained using 0.1 M Tris/HCl buffer containing 0.5 M NaSCN, High desorption rations(up to 98% of the adsorbed HSA) were observed. It was possible to reuse Cu(II) chelated-p(HEMA-co-MAH) beads without significant decreases in the adsorption capacities.

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Basic study on high gradient magnetic separation of nano beads using superconducting magnet for antibody purification

  • Jeongtae Kim;Insung Park;Gwantae Kim;Myunghwan Sohn;Sanghoon Lee;Arim Byun;Jin-sil Choi;Taekyu Kim;Hongsoo Ha
    • Progress in Superconductivity and Cryogenics
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    • v.25 no.4
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    • pp.60-64
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    • 2023
  • The manufacturing process of antibody drugs comprises two main stages: the upstream process for antibody cultivation and the downstream process for antibody extraction. The domestic bio industry has excellent technology for the upstream process. However, it relies on the technology of foreign countries to execute downstream process such as affinity chromatography. Furthermore, there are no domestic companies capable of producing the equipment for affinity chromatography. High gradient magnetic separation technology using a high temperature superconducting magnet as a novel antibody separation and purification technology is introduced to substitute for the traditional technology of affinity chromatography. A specially designed magnetic filter was equipped in the bore of the superconducting magnet enabling the continuous magnetic separation of nano-sized paramagnetic beads that can be used as affinity magnetic nano beads for antibodies. To optimize the magnetic filter that captures superparamagnetic nanoparticles effectively, various shapes and materials were examined for the magnetic filter. The result of magnetic separation experiments show that the maximum separation and recovery ratio of superparamagnetic nanoparticles are 99.2 %, and 99.07 %, respectively under magnetic field (3 T) and flow rate (600 litter/hr).

Affinity Apheresis for Treatment of Bacteremia Caused by Staphylococcus aureus and/or Methicillin-Resistant S. aureus (MRSA)

  • Mattsby-Baltzer, Inger;Bergstrom, Tomas;Mccrea, Keith;Ward, Robert;Msc, Lars Adolfsson;Larm, Olle
    • Journal of Microbiology and Biotechnology
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    • v.21 no.6
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    • pp.659-664
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    • 2011
  • Staphylococcus aureus (SA) bacteremia is associated with high mortality, and often results in metastatic infections. The methicillin-resistant SA (MRSA) is an urgent health care issue, as nosocomial infections with these bacteria represent limited treatment alternatives. Samples of whole blood containing challenge inoculums of SA and MRSA strains were passed through columns packed with surfaceheparinized polyethylene beads. The bound bacteria were eluted and quantitatively determined by culturing and by real-time PCR. Significant amounts of both SA and MRSA adhered to the heparinized beads (more than 65% of inoculated bacteria). After rinsing with buffer at high ionic strength, viable bacteria or bacterial DNA were eluted from the columns, indicating that the binding was specific. The conclusions that can be made from these experiments are that, as earlier reported in the literature, the high affinity of SA to heparin is retained in whole blood, and MRSA in whole blood binds to heparin with similar or higher affinity than SA. It should be possible to lower the amount of SA and/or MRSA from the blood of infected patients to levels that could be taken care of by the immune system. In previous studies, we have shown that passing blood from septic patients over beads coated with end-point-attached, biologically active heparin is a useful technique for regulating the levels of heparinbinding cytokine. These findings in combination with the present findings indicate the possibility of creating an apheresis technology for treatment of sepsis caused by SA and/or MRSA.

Affinity Separations Using Microfabricated Microfluidic Devices: In Situ Photopolymerization and Use in Protein Separations

  • Chen Li;Lee, Wen-Chien;Lee, Kelvin H.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.4
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    • pp.240-245
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    • 2003
  • The use of microfabricated microfluidic devices offers significant advantages over current technologies including fast analysis time and small reagent requirements. In the context of proteomic research, the possibility of using affinity-based separations for prefractionation of samples using microfluidic devices has significant potential. We demonstrate the use of microscale devices to achieve affinity separations of proteins using a device fabricated from borosilicate glass wafers. Photolithography and wet etching are used to pattern individual glass wafers and the wafers are fusion bonded at 650$^{\circ}C$ to obtain enclosed channels. A polymer has been successfully polymerized in situ and used either as a frit for packing beads or, when derivatized with Cibacron Blue 3GA, as a separation matrix. Both of these technologies are based on in situ UV photopolymerization of glycidyl methacrylate (GMA) and trimethylolpropane trimethacrylate (TRIM) in channels.

The Method Development for Biomarker Diagnosis Based on the Aptamer-protein Crosslink (앱타머와 단백질간 가교를 이용한 바이오마커 진단 방법 개발)

  • Lee, Bo-Rahm;Kim, Ji-Nu;Kim, Byung-Gee
    • KSBB Journal
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    • v.26 no.4
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    • pp.352-356
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    • 2011
  • The detection of biomarkers is an important issue for disease diagnosis. However, many systems are not suitable to detect the biomarker itself directly. For direct detection of biomarker proteins in human serum, a new affinity-capture method using aptamers combined with the mass spectrometry was suggested. Since signals from protein samples cannot be amplified, modified chromatin immunoprecipitation (ChIP) and subsequent cross-linking with formaldehyde between aptamers and target proteins were used not to lose the captured target proteins, which allowed us to perform a harsh washing step to remove the non-specifically bound proteins. As a model system, a thrombin aptamer was used as a bait and thrombin as a target protein. Using our modified ChIP and affinity-capture method, non-specific binding proteins on the beads decreased significantly, suggesting that our new method is efficient and can be applied to developing diagnosis systems for various biomarkers.

Picomolar Scale Determination of Carbohydrates Covalently Immobilized on Activated Beads Using Hydroxyl Functionality

  • Yu, Jae-Hoon;Chun, Sung-Min;Park, Ho-Koon;Park, Yong-Keun;Jeong, Sun-Joo
    • BMB Reports
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    • v.32 no.1
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    • pp.98-102
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    • 1999
  • Since carbohydrates are major mediators in cell-to-cell adhesion and communication, the development of specific and strong binders against them could generate promising therapeutics. As the first step towards that goal, sugar molecules have to be immobilized to be used as an affinity matrix. The amino functionality in sugar is the most active nucleophile for the immobilization, if the amino group is available. An alternative and general method is to use the hydroxyl group as a direct nucleophile, but the quantitation of immobilized hydroxyl groups is not easily done. To overcome this limitation, we have developed a method to immobilize various isomers of monosaccharides with p-nitrophenyl groups to the beads by using their hydroxyl groups. It was found that the amount of immobilized sugar was independent of the structure of the sugar, but was dependent on the number of hydroxyl groups. We also developed a sensitive method to quantify the amount of immobilized sugar at the picomolar scale by utilizing commercially available glycosidases to release a sensitive reporter molecule, p-nitrophenol, and detect it by HPLC. This new technique would allow a facile quantitation method for immobilized sugar molecules, which could be used as the affinity matrix to develop strong binders against biologically important sugars.

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Heavy Metal Removal Capacity of Chemically Modified Alginic Acid (화학적으로 개질된 알긴산의 중금속 제거능)

  • Lee, Soon-Hong;Kim, Kwang-Kook;Lee, Sang-Hoon
    • Journal of Korean Society on Water Environment
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    • v.21 no.6
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    • pp.569-574
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    • 2005
  • In this study, alginic acid that had an high affinity for a heavy metal and was noted for biological adsorbent was modified by an oxidizer, $KMnO_4$. Chemical modification changed hydroxyl of the alginic acid into carboxyl and compare with alginic acid, modified alginic acid exhibited a characteristics that carboxyl groups are comparatively high. For the use of them as an adsorbent, beads were prepared by dropping alginic acid and modified alginic acid solution in dilute 2 wt% $CaCl_2$ solution for non water soluble. The amount of removed $Cu^{2+}$ and $Pb^{2+}$ by modified alginic acid beads showed 84.7 mg and 90.9 mg per gram of beads, respectively. And it showed the amount of adsorbed heavy metal ions 10~20% higher than that of alginic acid beads in range of pH 4~7. In particular, modified alginic acid have a good adsorption capacity for $Cu^{2+}$ and $Pb^{2+}$ by Freundlich adsorption isotherm. According to this study, it is verified that alginic acid that is a nature high molecular substance improved capacity for actual application by increased heavy metal adsorption capacity by chemical modification.

Purification of the Candida utilis Extracellular Invertase using Affinity Chromatography

  • Ginalska, G.;Belcarz, A.;Lobarzewski, J.;Leonowicz, A.;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • v.30 no.3
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    • pp.12-17
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    • 2002
  • The extracellular invertase (EC 3.2.1.26) (Candida utilis) preparation was obtained from the liquid medium after desalting and freeze drying. This prepared enzyme was used for the comparative purification on 4 activated matrices by liquid column affinity chromatography method. In this method there were used controlled porous glass (CPG) silanized covalently activated by keratin, silanized silica gel and silica gel covalently covered by keratin. It was found that the invertase purification process was better using both CPG matrices (silanized CPG and keratin activated CPG) than these with two silica gel supports. Also the elution coefficient of the invertase from the two CPG columns was about 93 to 94%. Two silica gel supports found to be superior in terms of purification efficiency. The invertase purification process was confirmed by PAGE electrophoresis.

Antibacterial Activity of Essential Oils on the Growth of Staphylococcus aureus and Measurement of their Binding Interaction Using Optical Biosensor

  • Chung, Kyong-Hwan;Yang, Ki-Sook;Kim, Jin;Kim, Jin-Chul;Lee, Ki-Young
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1848-1855
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    • 2007
  • Antibacterial activity of essential oils (Tea tree, Chamomile, Eucalyptus) on Staphylococcus aureus growth was evaluated as well as the essential oil-loaded alginate beads. The binding interactions between the cell and the essential oils were measured using an optical biosensor. The antibacterial activity of the essential oils to the cell was evaluated with their binding interaction and affinity. The antibacterial activity appeared in the order of Tea Tree>Chamomile>Eucalyptus, in comparison of the inhibition effects of the cell growth to the essential oils. The association rate constant and affinity of the cell binding on Tea Tree essential oil were $5.0{\times}10^{-13}\;ml/(CFU{\cdot}s)$ and $5.0{\times}10^5\;ml/CFU$, respectively. The affinity of the cell binding on Tea Tree was about twice higher than those on the other essential oils. It might be possible that an effective antibacterial activity of Tea Tree essential oil was derived from its strong adhesive ability to the cell, more so than those of the other essential oils.