• Title/Summary/Keyword: G-BSA

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A comparison of detection capabilities of anti-IgG immobilizedby protein G and NHS (Protein G와 NHS를 이용하여 고정한 anti-IgG의 검출 성능 비교)

  • Sin, Eun-Jung;Sohn, Young-Ho;Sohn, Young-Soo
    • Journal of Sensor Science and Technology
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    • v.19 no.2
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    • pp.142-148
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    • 2010
  • We have compared and investigated the detection capabilities of antibody of immunoglobulin G(anti-IgG) immobilized by protein G and N-hydroxysuccinimide(NHS) at the end of the self-assembled monolayer(SAM). Surface plasmon resonance(SPR) sensor has been utilized to measure the interaction between biomolecules. After formation of the protein G and SAM, anti-IgG, bovine serum albumin(BSA) and IgG has been sequently injected. Through the reponse of the SPR, we can conclude that the protein G immobilized anti-IgG better than the SAM. In addition, IgG detection capability of the anti-IgG immobilized by the protein G showed better performance compared with that immobilized by the SAM.

Anti-allergic Effect of Eckolona cava Ethyl Acetate Fraction of on IgE/BSA-stimulated Bone Marrow-derived Cultured Mast Cells (IgE/BSA가 자극한 골수유래 비만 세포에 대한 감태 Ethyl Acetate 분획물의 항알러지 효능)

  • Han, Eui Jeong;Kim, Hyun Soo;Shin, Eun Ji;Kim, Min Ju;Han, Hee-Jin;Jeon, You-Jin;Jee, Youngheun;Ahn, Ginnae
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.277-284
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    • 2018
  • In this study, we investigated the anti-allergic effect of the ethyl acetate fraction of Ecklonia cava (EC-EtoAc) on the immunoglobulin E (IgE)/bovine serum albumin (BSA)-mediated activation of bone marrow-derived cultured mast cells (BMCMCs). We revealed that the $62.5{\mu}g/ml$ of EC-fractions ($EC-CHCl_3$, EC-Hexane and EC-EtoAc) inhibited IgE/BSA-activated ${\beta}$-hexosaminidase release from BMCMCs without cytotoxicity. Especially, EC-EtoAc showed the higher ${\beta}$-hexosaminidase release than the others. Also, EC-EtoAc reduced the expression levels of cytokines such as interleukin (IL)-$1{\beta}$, IL-4, IL-5, IL-6, IL-10, IL-13, interferon (IFN)-${\gamma}$ and tumor necrosis factor (TNF)-${\alpha}$ and a chemokine, thymus- and activation-regulated chemokine (TARC), compared to the only IgE/BSA-treated BMCMCs. Furthermore, EC-EtoAc significantly prevented the binding of IgE to Fc epsilon receptor $(Fc{\varepsilon}R)I$ and reduced the $Fc{\varepsilon}RI$ expression on the sensitized BMCMCs. Taken together, these results suggest that E. cava may be the natural agent with beneficial potentials for the treatment of type I allergic diseases induced by mast cell activation.

Effect of 65 kDa Buffalo Placental Protein on B-Cell Proliferation and Antibody Response

  • Deepthi, K.;Rajput, Y.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.3
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    • pp.432-437
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    • 2002
  • Immunosuppressive potential of 65 kDa buffalo placental protein (bPP65) on B-cell proliferation in vitro and antibody response in vivo was evaluated. B-cell proliferation was estimated by measuring incorporation of tritiated thymidine in buffalo lymphocytes while primary antibody responses against phytohaemagglutinin (PHA) or keyhole limpet haemocyanin (KLH) were evaluated in mice. bPP65 suppressed proliferation of lipopolysaccharide (a B-cell specific mitogen)-stimulated buffalo lymphocytes in vitro indicating suppression of B-cells. This suppression was dose dependent over the protein concentration range $25-100 {\mu}g/ml$. Primary antibody responses in mice against PHA and KLH in presence of bPP65 were lower as compared to in its absence but these were not statistically significant. Amino acid composition data of bPP65 and BSA suggested that bPP65 is different from BSA.

Effects of High Glucose and Advanced Glycosylation Endproducts(AGE) on the in vitro Permeability Model (당과 후기당화합물의 생체 외 사구체여과율 모델에 대한 역할)

  • Lee Jun-Ho;Ha Tae-Sun
    • Childhood Kidney Diseases
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    • v.10 no.1
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    • pp.8-17
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    • 2006
  • Purpose : We describe the changes of rat glomerular epithelial cells when exposed to high levels of glucose and advanced glycosylation endproducts(AGE) in the in vitro diabetic condition. We expect morphological alteration of glomerular epithelial cells and permeability changes experimentally and we may correlate the results with a mechanism of proteinuria in DM. Methods : We made 0.2 M glucose-6-phsphate solution mixed with PBS(pH 7.4) containing 50 mg/mL BSA and pretense inhibitor for preparation of AGE. As control, we used BSA. We manufactured and symbolized five culture dishes as follows; B5 - normal glucose(5 mM) + BSA, B30 - high glucose(30 mM) + BSA, A5 - normal glucose(5 mM) + AGE, A30 - high glucose(30 mM) + AGE, A/B 25 - normal glucose(5 mM) + 25 mM of mannitol(osmotic control). After the incubation period of both two days and seven days, we measured the amount of heparan sulfate proteoglycan(HSPG) in each dish by ELISA and compared them with the B5 dish at 2nd and 7th incubation days. We observed the morphological changes of epithelial cells in each culture dish using scanning electron microscopy(SEM). We tried the permeability assay of glomerular epithelial cells using cellulose semi-permeable membrane measuring the amount of filtered BSA through the apical chamber for 2 hours by sandwich ELISA. Results : On the 2nd incubation day, there was no significant difference in the amount of HSPG between the 5 culture dishes. But on the 7th incubation day, the amount of HSPG increased by 10% compared with the B5 dish on the 2nd day except the A30 dish(P<0.05). Compared with the B5 dish on the 7th day the amount of HSPG in A30 and B30 dish decreased to 77.8% and 95.3% of baseline, respectively(P>0.05). In the osmotic control group (A/B 25) no significant correlation was observed. On the SEM, we could see the separated intercellular junction and fused microvilli of glomerular epithelial cells in the culture dishes where AGE was added. The permeability of BSA increased by 19% only in the A30 dish on the 7th day compared with B5 dish on the 7th day in the permeability assay(P<0.05). Conclusion: We observed not only the role of a high level of glucose and AGE in decreasing the production of HSPG of glomerular epithelial cells in vitro, but also their additive effect. However, the role of AGE is greater than that of glucose. These results seems to correlate with the defects in charge selective barrier. Morphological changes of the disruption of intercellular junction and fused microvilli of glomerular epithelial cells seem to correlate with the defects in size-selective barrier. Therefore, we can explain the increased permeability of glomerular epithelial units in the in vitro diabetic condition.

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Antioxidative Effect of Ethanol Extract for 5 Kinds of Spice (5종의 향신료 에탄올 추출물의 항산화 효과)

  • Kim, Jin;Kim, Sung-Ae;Yun, Won-Kyung;Kim, Eun-Jeong;Woo, Mee-Kyung;Lee, Mee-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.9
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    • pp.1426-1431
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    • 2004
  • This study was performed to investigate the antioxidative effect of ethanol extracts of 5 spices. They were separately extracted in ethanol from dried samples at room temperature, and freeze-dried. In vitro testing were conducted by DPPH (1,1-diphenyl-2-picryl hydrazyl) radical scavenging activity, inhibition of iron-induced linoleate peroxidation and the inhibition of malondialdehyde (MDA) and bovine serum albumin (BSA) conjugation reaction. The ethanol extracts of clove (92.9%) and cinnamon (89.9%) showed the most effective results among five spices in the DPPH radical scavenging capacities. The inhibition rate of ethanol extract of clove on the lipid peroxidation was 55.8%. The ethanol extracts of mustard, wasabi and black pepper were effective in the inhibition of MDA and BSA conjugation reaction showing 73.2%, 72.2% and 61.6%, respectively. These results suggest that five spices tested in this study may enhance the antioxidative capacity, although the results were different according to the assay method and sample.

Studies on the Maturation of rabbit Follicular Oocytes in Vitro: Effects of Amino Acids and Carbohydrates

  • Bae, In-Ha
    • The Korean Journal of Zoology
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    • v.18 no.4
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    • pp.181-196
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    • 1975
  • Rabbit follicular oocytes were cultured in a basic medium containing 0.4% bovine serum albumin (BSA), carbohydrates and amino acids in various combinations. Osmolarity of the medium was maintained at 308 mOsm. The carbohydrates, pyruvate, lactate and glucose were all about equally beneficial, but not essential for rabbit oocyte maturation. Glutamine and proline, but not methionine or phenylalanine stimulated oocyte develoment. Glutamine stimulated more follicular oocytes to develop to prophase and metaphase II than did any of the three carbohydrates tested alone or in combination. Ammonia production after 24 hours of culture was highest in medium containing glutamine(15.2$\\mu$g/ml) but this was not inhibitory to maturation. Negligible amounts of ammonia were found with the other amino acids added. The optimum level of osmolarity for rabbit oocyte maturation appears to be ranged from 250$\\sim$310 mOsm with the maximum level of 270 mOsm. With 0, 0.08, 0.4, 2, 10 and 50 mM of glutamine in the medium, plus BSA but without carbohydrates, 30, 73, 70, 71, 59, 45% of the oocytes developed to prophase or metaphase II respectively. This indicates that no carbohydrate is required of the maturation of rabbit oocytes when 0.08$\\sim$2 mM of glutamine is included, which are the optimum range. Follicular oocytes could develop in the medium containing $^14 C$-glutamine and BSA but without carbohydrates or other organic compound. From the $^14 CO_2$ produced and TCA precipitable material isolated, it is suggested that glutamine probably is utilized by oocytes and cumulus cells as a source of energy as well as for protein synthesis.

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The Correlation of Antioxidative Effects of 5 Korean Common Edible Seaweeds and Total Polyphenol Content (한국산 5종 해조류의 항산화효과와 총 폴리페놀 함량과의 관련성)

  • Kwak, Chung-Shil;Kim, Sung-Ae;Lee, Mee-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1143-1150
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    • 2005
  • Antioxidative activities of 5 common edible seaweeds in Korea, three brown algae (seaweed fusiforme, sea mustard, sea tangle), one green algae (sea lettuce) and one red algae (laver), were examined. The antioxidative activities of ethanol extracts from these seaweeds were examined by measuring of inhibition rates against iron-induced linoleate peroxidation, DPPH (1,1-diphenyl -2-picrylhydrazyl) radical generation and MDA-BSA (malondialdehyde-bovine serum albumin) conjugation. Sea lettuce ethanol extract showed the strongest anti-oxidative activity among them, especially in inhibition against conjugation of lipid peroxide and protein. Second to sea lettuce, laver and sea tangle ethanol extracts showed high DPPH radical scavenging activity and inhibition against MDA-BSA conjugation. However, seaweed fusiforme and sea mustard ethanol extracts did not show antioxidative activities. Sea mustard contained the highest total flavonoids (11.33 mg/g dry wt) and sea lettuce contained the highest total polyphenol (8.97 mg/g dry wt) among these seaweeds. In addition, there was strong positive correlation between the antioxidative activity and total polyphenol content in these seaweeds, suggesting polyphenol compounds may contribute to antioxidative effect of seaweeds. From these data, it is suggested to consume much of seaweeds such as sea lettuce, laver and sea tangle to prevent age-related chronic diseases, and also develope neutraceutical products using polyphenol rich fraction from sea lettuce.

Effects of the Coptidis Rhizoma Extract on the Membranous Nephropathy induced by Cationic Bovine Serum Albumin in Mice (황연(黃連)이 Cationic Bovine Serum Albumin 투여로 유발된 Membranous Nephropathy Mouse Model에 미치는 영향)

  • Chae, Eun-Young;Cho, Chung-Sik;Kim, Cheol-Jung
    • The Korea Journal of Herbology
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    • v.24 no.1
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    • pp.99-110
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    • 2009
  • Objectives : The current treatment regimens for patients with nephrotic syndrome due to membranous nephropathy(MN) are based on steroids or immunosuppressive therapy with the aim of reducing proteinuria and improving outcome. Although these treatments attenuate the deterioration of renal function in MN patients, it has been suggested that all are burdened by significant toxicity. Therefore, more specific and less toxic therapies are needed. This study was to evaluate the effects of Coptidis Rhizoma Extract(CRE) on the MN induced by cBSA in mice. Methods : Mice were divided into 4 groups. One group named for 'Normal' was injected with a saline solution not to be immunized. The rest groups were treated as follows; After mice were immunized with 0.2 mg of cBSA and Freund's complete adjuvant one time every two weeks for 6 weeks, they received intra-peritoneal injection of 10 mg/kg of cBSA daily for 4 weeks. Also, they were divided into 3 groups. The first named for 'Control' was not given CRE. The second for 'CRE-250' was given oral administration of 250 mg/kg of CRE daily for 4 weeks. The third for 'CRE-500' was given 500 mg/kg of CRE. All of mice were sacrificed 4 weeks after the first immunization. We measured a body weight and 24hrs proteinuria as well as serological analysis. The morphologic changes of renal glomeruli were also observed with a light microscope and an electron microscope. Results : The levels of 24 hrs proteinuria, triglyceride, IgG, IL-6 were significantly decreased in both CRE groups. And the level of IgM was significantly decreased in CRE-250 group. In histological findings of kidney tissue, thickening of GBM and deposition of electron-density were consideraly decreased in both CRE groups. Conclusions : The present study suggests that CRE is highly effective when treating mice with MN induced by cBSA. More clinical data and studies are to be done for efficient application.

Detection of IgG Using Thiolated Protein G Modified SPR Sensor Chip (Thiolated protein G로 개질된 SPR 센서 칩을 이용한 IgG 검출)

  • Sin, Eun-Jung;Lee, Yeon-Kyung;Sohn, Young-Soo
    • Journal of Sensor Science and Technology
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    • v.20 no.6
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    • pp.434-438
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    • 2011
  • A portable surface plasmon resonance(SPR) based immunosensor using thiolated protein G and protein G was developed for the detection of immunoglobulin G(IgG). The protein G has specific affinity with Fc fragment of IgG and was thiolated by 2-Iminothiolane for introduction of thiol groups. Anti-IgG, bovine serum albumin(BSA), and IgG have been sequently injected after surface modification of gold sensor chip with protein G and thiolated protein G. The output signal was increased with the injection of each protein and the actual signal was measured by subtracting signal of reference channel from signal of sample injected channel. The experimental results showed the higher detection capability of IgG using thiolated protein G compared with protein G. From these results, we can conclude that the current surface modification technique and the portable SPR sensor system can be applied to various immunosensors for diagnosis.

Foam Separation of Bovine Serum Protein Fractions (소 혈청 단백질 분획들의 기포분리 현상에 관한 연구)

  • Lee, Boo-Yong;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.19 no.3
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    • pp.225-230
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    • 1987
  • The foam separation of bovine serum proteins was investigated and the protein fractionation by foam separation was analyzed by PAG electrophoresis. The protein concentration for the surface excess formation of bovine serum was in the range of $20-800\;{\mu}g/ml$. At pH 5, the foamate volume was maximum, but the enrichment ratio minimum. As the temperature was elevated, the foamate volume decreased and the enrichment ratio increase. As the gas flow rate increased from 25 to 100 ml/min, the foamate volume decreased and the enrichment ratio increased. The enrichment ration became maximum when the added ionic strength of serum solution was in the range of 1-3 by the addition of different types of salts, and this was related to the reduction of surface tension of the solution. In general, BSA, ${\alpha}_1$, and ${\alpha}_2-globulins$, which have relatively small molecular weight and high hydrophobicity, moved easily to the foam, and the separation of protein fractions in the serum varied with the changes in pH, temperature, gas flow rate and ionic strength of the solution.

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