• Title/Summary/Keyword: Bovine serum albumin

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Protein Adsorption on Ion Exchange Resin: Estimation of Equilibrium Isotherm Parameters from Batch Kinetic Data

  • Chu K.H.;Hashim M.A.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.1
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    • pp.61-66
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    • 2006
  • The simple Langmuir isotherm is frequently employed to describe the equilibrium behavior of protein adsorption on a wide variety of adsorbents. The two adjustable parameters of the Langmuir isotherm - the saturation capacity, or $q_m$, and the dissociation constant, $K_d$ - are usually estimated by fitting the isotherm equation to the equilibrium data acquired from batch equilibration experiments. In this study, we have evaluated the possibility of estimating $q_m$ and $K_d$ for the adsorption of bovine serum albumin to a cation exchanger using batch kinetic data. A rate model predicated on the kinetic form of the Langmuir isotherm, with three adjustable parameters ($q_m,\;K_d$, and a rate constant), was fitted to a single kinetic profile. The value of $q_m$ determined as the result of this approach was quantitatively consistent with the $q_m$ value derived from the traditional batch equilibrium data. However, the $K_d$ value could not be retrieved from the kinetic profile, as the model fit proved insensitive to this parameter. Sensitivity analysis provided significant insight into the identifiability of the three model parameters.

Inhibitory effects of curcumin on high glucose-induced damages: Implications for alleviating diabetic complications

  • Kim, Kyeong Yee;Kim, Choon Young
    • Food Science and Preservation
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    • v.24 no.4
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    • pp.536-541
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    • 2017
  • Hyperglycemia found in diabetes mellitus causes several physiological abnormalities including the formation of advanced glycation end products (AGEs) and oxidative stress. Accumulation of AGEs and elevation of oxidative stress plays major roles in the development of diabetic complications. Adiponectin secreted from adipocytes is known to improve insulin sensitivity and blood glucose level. Curcumin (CCM), a bioactive component of turmeric, has been reported as a potent antioxidant. Present work aimed to elucidate the roles of CCM in high glucose-induced protein glycation and intracellular events in mature adipocytes. The results demonstrated that CCM inhibited the formation of fluorescent AGEs by approximated 52% at 3 weeks of bovine serum albumin (BSA) glycation with glucose. Correspondingly, CCM decreased the levels of fructosamine and ${\alpha}-dicarbonyl$ compounds during BSA glycation with glucose. These data suggested that CCM might be a new promising anti-glycation agent. Also, CCM reduced high glucose-induced oxidative stress in a dose dependent manner, whereas CCM treatment time-dependently elevated the expression of adiponectin gene in 3T3-L1 adipocytes. The findings from this study suggested the possibility of therapeutic use of CCM for the prevention of diabetic complications and obesity-related diseases.

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.

Liquid Crystal Based Optical Sensor for Imaging Trypsin Activity at Interfaces Between Aqueous Phases and Thermotropic Liquid Crystals

  • Zhang, Minmin;Jang, Chang-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.10
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    • pp.2973-2977
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    • 2013
  • In this study, we developed a liquid crystal (LC)-based optical sensor for monitoring enzymatic activity through orientational changes in liquid crystals (LCs) coupled to the properties of a poly-${\small{L}}$-lysine (PLL)-based polymeric membrane. We prepared a PLL-based polymeric membrane at the planar interface between the thermotropic liquid crystal and aqueous phases. The PLL-based polymeric membrane was obtained by contacting the PLL solution with water immiscible LCs, 4-cyano-4'-pentyl-biphenyl (5CB) doped with adipoyl chloride. We then investigated the membrane properties by examining the permeability of the membrane to phospholipids, 1,2-didodecanoyl-rac-glycero-3-phosphocholine (DLPC). The permeability of the membrane to transport phospholipids was monitored through the orientational transition of 5CB in contact with the dispersions of DLPC. Since trypsin can enzymatically catalyze the hydrolysis of PLL, we incubated an aqueous trypsin solution with the membrane for 2 h at room temperature to cause an increase in the permeability of the polymeric membrane to DLPC. As a result, a bright to dark optical shift of LCs was observed, which implied that an enzymatic reaction between trypsin and PLL-based membrane occurred. Two control experiments using chymotrypsin and bovine serum albumin (BSA) revealed no sign of improved permeability based on the orientational transition of LCs.

Conformational Stability of Proteins in Colloidal Food Model System (콜로이드 모델 식품에 있어 단백질의 구조적 안정성)

  • Song, Kyung-Bin
    • Korean Journal of Food Science and Technology
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    • v.25 no.3
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    • pp.277-281
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    • 1993
  • To elucidate the conformational stability of proteins in colloidal food system, molecular properties of various proteins such as chemically modified ${\beta}-lactoglobulin$, bovine serum albumin (BSA) structural intermediates, and ${\beta}-casein$ under chaotropic conditions, were examined using circular dichroism, SS bond content, and hydrodynamic radius determination. As refolding time increases, BSA intermediates approach the conformation of native BSA. And succinylation made ${\beta}-lactoglobulin$ have more aperiodic structure by increasing net negative charge. Also, under chaotropic conditions, the conformation of P-casein was affected by hydrophobic interactions. This study clearly indicates that hydrophobic interactions and electrostatic interactions are major contributing factors in conformational stability of proteins.

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Effect on Immune Cells of Proteoglycan Originating from Rhanella aquatilis (Rhanella aquatilis 유래 당단백질의 면역세포에 미치는 영향)

  • Park, Hae-Gi;Kim, Kwang-Hyeon
    • Microbiology and Biotechnology Letters
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    • v.42 no.3
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    • pp.312-315
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    • 2014
  • So as to evaluate its use an immune stimulator in humans, the toxicity and action against immune cells by an anti-yeast substance (AYS), a bacterial proteoglycan, were investigated. The AYS did not possess hemolytic activity with human red blood cells (hRBC). Nor did it exhibit cytotoxicity against human peripheral blood mononuclear cells (hPBMC). In addition, the AYS did not induce hPBMC proliferation, but it did agglutinate hPBMCs in vitro. Moreover, hPBMC induced inflammatory cytokines such as IL-6, IL-5, IFN-${\gamma}$ and TNF-${\alpha}$ with the AYS during culture. Compared with alum, the AYS as an adjuvant has an increased antibody production rate against bovine serum albumin (BSA) in mice.

Characterization of Biocompatible Polyelectrolyte Complex Multilayer of Hyaluronic Acid and Poly-L-Lysine

  • Hahn, Sei-Kwang;Allan S. Hoffman
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.3
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    • pp.179-183
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    • 2004
  • A biocompatible polyelectrolyte complex multilayer (PECML) film consisting of poly-L-lysine (PLL) as a polycation and hyaluronic acid (HA) as a polyanion was developed to test its use for surface modification to prevent cell attachment and protein drug delivery. The formation of PECML through the electrostatic interaction of HA and PLL was confirmed by contact angle measurement, ESCA analysis, and HA content analysis. HA content increased rapidly up to 8 cycles for HA/PLL deposition and then slightly increased with an increasing number of deposition cycle. In vitro release of PLL in the PECML continued up to 4 days and ca. 25% of HA remained on the chitosan-coated cover glass after in vitro release test for 7 days. From the results, PECML of HA and PLL appeared to be stable for about 4 days. The surface modification of the chitosan-coated cover glass with PECML resulted in drastically reduced peripheral blood mononuclear cell (PBMC) attachment. Concerned with its use for protein drug delivery, we confirmed that bovine serum albumin (BSA) as a model protein could be incorporated into the PECML and its release might be triggered by the degradation of HA with hyaluronidase.

Positively Charged Silver Nanoparticles Threaded on Carbon Nanotube for the Efficient Delivery of Negatively Charged Biomolecules

  • Park, Hyung-Seok;Hwang, Ji-Young;Shin, Ueon-Sang;Kim, Hae-Won;Gong, Myoung-Seon
    • Bulletin of the Korean Chemical Society
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    • v.32 no.10
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    • pp.3581-3586
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    • 2011
  • Silver nanoparticle (Ag-NPs)-immobilized and amine-functionalized carbon nanotubes (MWCNTs), MWCNT-Ag-$NH_2$, were easily prepared in order to develop an efficient delivery system of biomolecules without complicated processes of manufacture. For this, Ag-NPs-immobilized MWCNTs, MWCNT-Ag, were initially prepared in order to create large surface area to enable more efficient linkage with guest-molecules using pristine MWCNTs. The Ag-NPs on MWCNTs were further positively functionalized with 2-aminoethanthiol to allow ionic linkage with biomolecules. Ultimately, the positively charged delivery system proved to be highly effective for the binding capacity of bovine serum albumin (BSA) as a negatively charged model protein, when compared to that of lysozyme used as a positively charged model protein. The releasing profile of BSA was observed in almost linear pattern for about two weeks in a saline solution. This study demonstrated the potential usefulness of the pristine MWCNTs in conjunction with Ag-NPs for the selective delivery of many (negatively or positively) charged biomolecules including proteins and genes.

A Possible Merge of FRET and SPR Sensing System for Highly Accurate and Selective Immunosensing

  • Lee, Jae-Beom;Chen, Hongxia;Lee, Jae-Wook;Sun, Fangfang;Kim, Cheol-Min;Chang, Chul-Hun L.;Koh, Kwang-Nak
    • Bulletin of the Korean Chemical Society
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    • v.30 no.12
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    • pp.2905-2908
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    • 2009
  • Immuno-sensing for high accurate and selective sensing was performed by fluorescence spectroscopy and surface plasmon resonance (SPR), respectively. Engineered assembly of two fluorescent quantum dots (QDs) with bovine serum albumin (BSA) and anti-BSA was fabricated in PBS buffer for fluorescence analysis of fluorescence resonance energy transfer (FRET). Furthermore, the same bio-moieties were immobilized on Au plates for SPR analysis. Naturally-driven binding affinity of immuno-moieties induced FRET and plasmon resonance angle shift in the nanoscale sensing system. Interestingly, the sensing ranges were uniquely different in two systems: e.g., SPR spectroscopy was suitable for highly accurate analysis to measure in the range of 10$^{-15{\sim}-10$ng/mL while the QD fluorescent sensing system was relatively lower sensing ranges in 10$^{-10{\sim}-6$ng/mL. However, the QD sensing system was larger than the SPR sensing system in terms of sensing capacity per one specimen. It is, therefore, suggested that a mutual assistance of FRET and SPR combined sensing system would be a potentially promising candidate for high accuracy and reliable in situ sensing system of immune-related diseases.

Identification of AGE-precursors and AGE formation in glycation-induced BSA peptides

  • Ahmad, Waqar;Li, Lili;Deng, Yulin
    • BMB Reports
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    • v.41 no.7
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    • pp.516-522
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    • 2008
  • The glycation of BSA leads to protein/peptide modifications that result in the formation of AGEs. AGEs react with the amino groups of N-terminal amino acid residues, particularly arginine and lysine residues. Enhanced AGE formation exists in the blood and tissues of diabetics, as well as in aging and other disorders. The Identification of AGEs is of great importance. Mass spectrometry has been applied to identify and structurally elucidate AGEs. Here, we report on the identification of AGE-peptides and AGE precursors based on relative mass changes as a result of specific AGE formation. HPLC-ESIMS, ESI-MS/MS, and the Mascot database were used. The relative mass changes due to the specific type of AGE formation were added to the identified peptides followed by a manual search of the glycated samples, which resulted in the identification of seven peptides for the formation of five AGEs, namely CML, pyrraline, imidazolone A, imidazolone B, and AFGP. Four glycated peptides (FPK, ECCDKPLLEK, IETMR, and HLVDEPQNLIK) were identified in the formation of AGE-precursors.