• Title/Summary/Keyword: Crossed immunoelectrophoresis

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Comparison of soluble antigens from Leptospira interrogans serovars by SDS-PAGE, Crossed Immunoelectrophoresis and Immunoblotting (SDS-PAGE, Crossed Immunoelectrophoresis 및 Immunoblotting을 이용한 Leptospira interrogans 혈청형간 항원 비교)

  • Baik, Yeong-ok;Mah, Jum-sool
    • Korean Journal of Veterinary Research
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    • v.32 no.2
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    • pp.195-205
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    • 1992
  • The soluble antigen profiles and antigenic specificities of Leptospira interrogans serovars icterhaemorrhagiae, canicola, pomona and hardjo were examined by SDS-polyacrylamide gel electrophoresis, crossed immunoeletrophoresis and immunoblotting. The profiles of protein, glycoprotein and fraction containing N-acetylglucosamine of 4 serovars were compared. The protein profiles of 4 serovars were very similar except the range of 14,400 to 30,000 daltons. Molecular weight of glycoprotein of L, pomona was lower than other serovars. L canicola showed extra N-acetylglucosamine bands having molecular weight of 82,000 and 90,000 daltons. In crossed immunoelectrophoresis, a close antigenic relationship was found between L icterohaemorrhagiae and L canicola. In immunoblottings conducted with soluble antigens and rabbit antisera of 4 serovars, Leptospira interrogans serovars possessed cross-reactive antigens and serovar-specific antigens. The molecular weights of serovar-specific antigens were 45,000, 82,000 and 90,000, 31,000 and 24,000 daltons in L icterohaemorrhagiae, L canicola, L pomona, and L hardjo, respectively.

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Soluble Proteins Analysis of Class Cephalopoda in the Yellow Sea(II) (황해산 두족류의 가용성 단백질에 대한 연구(II))

  • 허회권;황규린
    • Journal of Aquaculture
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    • v.10 no.4
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    • pp.425-433
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    • 1997
  • The isolated soluble eye proteins from six species (Sepia esculenta, Sepiella japonica, Loligo chinensis, Todarodes pacificus, Loligo beka and Ocotopus minor) of class Cephaloopoda were compared by crossed immunoelectrophoresis methods using antibodies directed against total soluble eye proteins antigens. It showed a distinct antigen-antibody reaction between the species of order Sepioidea and various antiserum of class Cephalopoda. Our results suggested that the common precipitation lines of Sepia and Loligo species which were different from that of Octopus minor. By which obtained from elution of gel filtration chromatography and 12.5% SDS-polyacrylamide gel electrophoresis, the molecular weight of Todarodes pacificus eye protein (crystallin) was determined in abut 20-35 KDa.

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Ovarian and Fat Body Yolk Protein Synthesis in Culex piplens pallens (홍모기(Culex pipiens pallens) 지방체와 난소에서의 난황단백질합성에 관한 연구)

  • 이승훈;박영민;성기창
    • The Korean Journal of Zoology
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    • v.36 no.3
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    • pp.416-424
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    • 1993
  • Ovarian Yolk protein (YP2) synthesis has been investigated in mosquito, Culex pipiens pallens. Yolk protein amount which was syntheized in fat body, accumulated into ovary were analyzed by Rocket immunoelectrophoresis and in vitro organ culture. The result was that yolk protein synthesis began to occur at 6hrs after blood meal, reached at maximum level by 24hrs, and was completed within 48hrs. Yolk protein accmulation into the ovary began to start at 6hrs and coutinued for up to 60hrs after blood meal. Extract from 0, 24, 48, 72hrs ovaries after blood meal were analyzed by electrophoresis and Western blotting. The result was that 24hrs ovary contain one yolk protein(YP1), and 48, 72hrs ovaries contain two kinds of yolk proteins(YPl and YP2). When 48hr ovaries and fat bodies were incubated in $^3$H-leucine contained medium, protein synthesis was not occurred in fat body, but ovary synthesized much protein contained yolk protein (YP2). The result of crossed immunoelectrophoresis represented the same immunity between YPl and YP2. The present data suggest that ovary synthesize yolk protein(YP2) in mosquito, Culex pipiens pallens.

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Characterization of Two Glucans Activating an Alternative Complement Pathway from the Fruiting Bodies of Mushroom Pleurotus ostreatus

  • Kweon, Mee-Hyang;Lim, Wang-Jin;Yang, Han-Chul;Sung, Ha-Chin
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.267-271
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    • 2000
  • Abstract Two glucans (PONGa and PONGb) differing in their anomeric and glycosidic linkage structures were isolated from the water-insoluble materials (PON) of Pleurotus ostreatus basidiocarps, which activated the complement system and were almost soley composed of D-glucose. The isolatIon was achieved by repeated precipitations with ethanol and adsorption on concanavalin A (Con A) of paN suspension in thymol/NaCL Based on methylation analysis. IR, GLC-MS, $^1H,{\;}and{\;}^{13}C-NMR$ spectroscopies, PONGa was found to be a branched a-glucan composed of ${\alpha}-linked$ D-glucopyranose residues and ${\alpha}-linked$ units with 6-branching points, whereas PONGb was a linear ${\beta}-1,3-glucan$ composed mainly of ${\beta}-1,3-linked$ D-glucopyranose residues. The PONGb particles reacted more potently than the PONGa particles as C3 activator in alternative complement hemolysis and crossed-immunoelectrophoresis using anti-human C3, thereby suggesting that the complement activating components of PON were ${\beta}-(13)-glucans rather$ than ${\alpha}-glucan$ components.onents.

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Immunomodulating Activity of the Exopolymer from Submerged Mycelial Culture of Phellinus pini

  • Jeong, Sang-Chul;Cho, Sung-Pill;Yang, Byung-Keun;Jeong, Yong-Tae;Ra, Kyung-Soo;Song, Chi-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.15-21
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    • 2004
  • The immunomodulating activities and chemical characteristics of a water-soluble exopolymer from submerged mycelial culture of Phellinus pini were studied. Anticomplementary activity of this polymer was found to be $73.2\%$, and its activation system occurred through both classical and alternative pathways, where the classical pathway was detected to be the major one by crossed immunoelectrophoresis. Nitric oxide (NO) release ability and acid phosphatase activity of macrophage were increased by 1.6-fold ($100{\mu}g/ml$) and 3.4-fold ($500{\mu}g/ml$), respectively, and splenocyte proliferation in mixed lymphocyte reaction (MLR) was also increased by 2.6-fold ($200{\mu}g/ml$), compared to the control. The molecular weight of this polymer, determined by HPLC, was under 5 kDa. Total sugar and protein contents were 89.7 and 10.3%, respectively. Both sugar and amino acid compositions of the exopolymer were also analyzed.

Isolation and Characterization of Acidic Polysaccharides Activating Complement System from the Hot Water Extracts of Pteridium aquilinum var. latiusculum (고사리 열수 추출물로부터 보체계 활성화 산성 다당의 분리 및 특성)

  • O, Byeong-Mi;Gwon, Mi-Hyang;Na, Gyeong-Su
    • The Korean Journal of Food And Nutrition
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    • v.7 no.3
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    • pp.159-168
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    • 1994
  • From the hot water extract of bracken(Pteridium aquilinum var. latiusculum), a Korean win edible plant, anti-complementary acidic polysaccharides were Isolated. Crude polysaccharide fraction(HPA-1) was obtain ed by methanol reflux, ethanol precipitation, dialysis, and lyophilization. HPA-1 contained 81.80% of total sugar, 30.40% of uronic acid, and 15.60cA of protein. HPA 1 was purified consecutively by cetavlon fractionation and chromatography including ion exchange nth DEAE-Sepharose CL 6B and gel permeation with Sephadex G-100 and Sepharose CL-6B. HPA-2- IVa and HPA-Va-2 were nearly homogeneous on HPLC and had 500,000 and 560,000 daltons of molecular weights, respectively. HPA-2-Wa consisted of fucose, galacturonic acid, and glucuronic acid at the molar ratio of 1.40 : 0.97 : 1.88. HPA-2-Va 2 was composed of rhamnose, galactose, and galacturonic acid at the molar ratio of 1.00 : 1.38 : 1.39. The polysaccharides were found to activate the C3 component both In the presence and In the absence of Ca2+ through the crossed-immunoelectrophoresis suggesting that those Involved in both classical and alternative complement pathway.

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Isolation and Characterization of Biopolymers Extracted from the Bark of Acanthopanax sessiliflorus and Their Anticomplement Activity

  • Jeong Sang-Chul;Yang Byung-Keun;Jeong Yong-Tae;Rao Koyyalamudi Sundar;Song Chi-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.21-28
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    • 2007
  • The crude biopolymer (AS-S1) and endobiopolymer (AS-S2) were isolated from the dry stem bark of Acanthopanax sessiliflorus and tested for anti complement activity. The two potent anticomplement biopolymers, AS-1 and AS-2-Fr.I, were isolated by the combination of ion-exchange chromatography and gel filtration methods from the endo-biopolymers (AS-S2). The anticomplement activity of AS-1 (MW 12 kDa) and AS-2-Fr.I (MW 180 kDa) were found to be 84.4% and 100.0%, respectively, at the concentration of $25{\mu}g/ml$. Activated pathway of the complement system occurred in both classical and alternative pathways, as evidenced by crossed immunoelectrophoresis(CIEP), where a major pathway was detected to be the classical one. It was found that the anticomplement activities of the periodate oxidized were decreased significantly, but those of pronase digested biopolymers of AS-1 and AS-2-Fr.I were decreased very little. The AS-1 contained 2,4,6-tri-O-methyl-D-glucitol, 2,3,6-tri-O-methyl-D-galacitol, and 2,3,6-tri-O-methyl-D-galacitol, which indicated that AS-1 contained a $(1{\rightarrow}3),\;(1{\rightarrow}4)-linked$ glucopyranosyl residue and a $(1{\rightarrow}4)-linked$ galactosyl residue. AS-2-Fr.I contained mainly 2,4-di-O-methyl-D-mannitol and 2,3,4-tri-O-methyl-D-galacitol, which contained $(1{\rightarrow}3),\;(1{\rightarrow}6)$ linked mannosyl and $(1{\rightarrow}6)$ linked galactosyl residues.

Effects of Alkali Extract of Ganoderma lucidum IY007 on Complement and Reticuloendothelial System (영지 균사체의 알칼리 추출물이 보체계와 망내계에 미치는 영향)

  • Lee, June-Woo;Jeong, Hoon;Chung, Chun-Hee;Lee, Kweon-Haeng
    • The Korean Journal of Mycology
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    • v.18 no.3
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    • pp.137-144
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    • 1990
  • To examine effects on complement and reticuloendothelial system, alkali extract was isolated from cultured mycelium of Ganoderma lucidum IY107. It was shown to strongly activate both classical and alternative pathways of complement as compared with krestin. Activated complement C3, 3rd peak, was observed by crossed immunoelectrophoresis. It was also shown to activate reticuloendotherial system of ICR mice in the carbon clearance test and to increase hemolytic plaque forming cells of the spleen. Carbohydrate and protein contents of the alkali extract were 10% and 49%, respectively. The carbohydrate consisted of four monosaccharides and the protein contained 16 amino acids.

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Immunopotentiating Activities of Cellular Components of Lactobacillus brevis FSB - 1 (Lactobacillus brevis FSB - 1의 균체성분에 의한 면역증진 활성)

  • Kim, Seong-Yeong;Shin, Kwang-Soon;Lee, Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.9
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    • pp.1552-1559
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    • 2004
  • In order to evaluate the potential utilization value as a novel probiotic strain, the immunopotentiating activities of the cellular components from Lactobacillus brevis FSB-1 were examined. L. brevis FSB-1 isolated from kimchi were fractionated into the whole cell, cell wall, cytosol and extracellular preparation, and each fraction was examined on intestinal immune system modulating activity in vitro. The cell wall and cytosol preparation showed the relatively high bone marrow cell proliferating activity through Peyer's patch cell in a dose-dependent manner. But these preparations did not directly stimulate the bone marrow cell proliferation. The whole cell, cell wall and cytosol preparation also induced considerable levels of macrophage activation and mitogenicity of murine splenocytes in vitro. The anti-complementary activity (ITCH_(50)) of the cytosol fraction of L. brevis FSB-1 was the most potent in the cellular components, and the activity showed dose dependency. The complement activation by the cytosol fraction of L. brevis FSB-1 occurs via both alternative and classical pathways, which confirmed by the crossed immunoelectrophoresis using anti-human C3.

Characterization of Anti-Complementary Polysaccharides Isolated from Fruit Wine Using Korean Pears (배를 이용한 과실주로부터 분리한 항보체 활성화 다당의 특성)

  • Choi, Jung-Ho;Shin, Kwang-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.63-69
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    • 2011
  • To characterize the polysaccharides which exist as soluble forms in Korean traditional alcoholic beverages, the polysaccharides were isolated from Korean pear wine and their anti-complementary activities were examined. The main polysaccharide, PW-1 was purified to homogeneity from the crude polysaccharide (PW-0) in pear wine by size exclusion chromatography using Sephadex G-75. Molecular mass of PW-1 was estimated to be 150 kDa and it contained significant proportion of mannose (81.8%) and 5 different minor component sugars such as arabinose (1.2%), galactose (2.7%), glucose (8.5%), galacturonic acid (5.3%) and glucuronic acid (0.5%). These analyses indicated that the main polysaccharide in pear wine was mainly present as a mannan which had originated from the cell walls of fermenting yeasts. On the other hand, PW-1 showed potent anti-complementary activity in a dose-dependent fashion. Identification of C3 activation products by the crossed immunoelectrophoresis using anti-human C3 and anti-complementary activity of PW-1 in $Ca^{++}$-free condition suggested complement activations by PW-1 from Korean pear wine occur via both classical and alternative pathways.