[Comparative immunologic and physico-chemical properties of neurospecific antigens--bull antigen D and rat antigen alpha].
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Immunological and physicochemical properties were compared for water-soluble acid neurospecific antigens: D of the bull and alpha of the rat. During immunoelectrophoresis in agar gel antigen D migrates as two immunologically indentical fractions having a position of prealbumins and alpha-globulins of blood serum. Antigen D from the bull brain is incompletely immunologically identical with antigen alpha from the rat brain. The molecular weight of antigen D determined under conditions of Sephadex G-100 column gel chromatography is 73000.Keywords:
Sephadex
Immunoelectrophoresis
Immunodiffusion
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Sephadex
Immunodiffusion
Immunoelectrophoresis
Excretory system
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Immunoelectrophoresis
Immunodiffusion
Sephadex
Ouchterlony double immunodiffusion
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A cell-mediated immunologic granulomatous response to Schistosoma mansoni eggs is now generally accepted as being responsible for the hepatosplenic disease of chronic schistosomiasis. Previous investigations have demonstrated that a soluble extract of S. mansoni eggs (SEA) both induces and elicits granulomatous hypersensitivity and other forms of cell-mediated immunologic reactivity. Mice with chronic light S. amnsoni infections show spontaneous suppression of granulomatous hypersensitivity in the presence of high levels of anti-SEA antibodies. Immunodiffusion analysis using antiserum obtained from these mice and SEA resulted in the identification of three major serologic antigens which have been designated MSA1, MSA2, and MSA2. Initial studies with Sephadex G-200 gel filtration and polyacrylamide gel electrophoresis indicated that the three antigens were markedly different in m.w. and at least two of the three were glycoproteins. The antigens were then extracted from crude SEA by adsorption to a concanavalin A Sepharose affininity column. The eluted antigens were separated from each other by ion exchange chromatography on DEAE cellulose. On polyacrylamide gel electrophoresis (PAGE) MSA1 and MSA2 were homogenous; MSA3 was estimated to be 70% pure. The purified antigens were radiolabeled and were passed through Sephadex G-200 or Bio-Gel A 1.5 columns to determine their m.w. These studies have shown that MSA1 and MSA2 are glycoproteins of m.w. 137,000 and 465,000 daltons, respectively; the m.w. of MSA3 is in the range of 50,000 to 70,000 daltons. PAGE of the purified antigens revealed the following R.F.s: MSA1=0.34, MSA2=0.20, and MSA3=0.48. MSA1 and MSA2 were employed in radioimmunoassays using the ammonium sulfate method of Farr. On the basis of immunodiffusion analysis and radioimmunoassay, MSA1 exhibits a degree of stage and species specificity consistent with the granulomatous response to S. mansoni eggs. The potential specificity of the MSA1 radioimmunoassay and its great sensitivity suggest a role in the immunodiagnosis of schistosomiasis.
Sephadex
Immunodiffusion
Ouchterlony double immunodiffusion
Immunoelectrophoresis
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Immunodiagnostic potential fractions in fluke antigens and excretory/secretory (E/S) antigen of F. gigantica have been identified using double immunodiffusion (DID), immunoelectrophoresis (IEP) and sodium dodecyl sulphate (SDS) PAGE electrophoresis. Hyperimmunised rabbits elicited considerable level of antibody response with few precipitin lines showing reaction of identity in DID suggesting presence of some common fractions in all antigens. Mature fluke somatic (MFSAg) and immature fluke somatic (IMFSAg) antigens showed exactly similar pattern of precipitin lines in both DID and IEP. Antibodies to experimentally infected rabbits with F. gigantica were also detected by DID using MFSAg and E/S antigens at 4 weeks post infection. However, out of 6-7 fractions obtained from Sephadex G-200 column chromatography in MFSAg and IMFSAg antigens only F3, F4 and F5 along with E/S antigen were able to detect antibodies by DID. Mature fluke and E/S antigen separated out in 9 bands in the range of 12 to 95 kDa and 5 bands in the range of 12 to 29 kDa respectively of which proteins of molecular weight 12, 15, 18, 28 and 29 kDa were common in both antigens. Therefore, it appears that immunodiagnostic fractions of antigens may be present in between 12 and 29 kDa.
Precipitin
Immunoelectrophoresis
Fasciola gigantica
Sephadex
Somatic antigen
Immunodiffusion
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Immunological and physicochemical properties were compared for water-soluble acid neurospecific antigens: D of the bull and alpha of the rat. During immunoelectrophoresis in agar gel antigen D migrates as two immunologically indentical fractions having a position of prealbumins and alpha-globulins of blood serum. Antigen D from the bull brain is incompletely immunologically identical with antigen alpha from the rat brain. The molecular weight of antigen D determined under conditions of Sephadex G-100 column gel chromatography is 73000.
Sephadex
Immunoelectrophoresis
Immunodiffusion
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The human heart antigens demonstrated in citric acid extracts by immunodiffusion were submitted to further analysis. Heart antigens were precipitated at 30% to 70% saturated ammonium sulphate. These fractions contained antigens reactive with rabbit antisera to human heart. The third fraction out of four proteins which were separated by Sephadex G-150 gel filtration, was reactive against rabbit antisera to human heart. The intensely stained proteins of both heart antigens were located at the nineth protein band out of proteins separated by polyacrylamide gel electrophoresis. This protein exhibited precipitin line against rabbit antisera to human heart by two-dimensional electrophoresis, and had mobility between that of serum alpha2 and beta1 globulin in immunoelectrophoresis.
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Banach , T. M. (University of Manitoba, Winnipeg, Canada), and R. Z. Hawirko . Isolation and characterization of two antigens of Corynebacterium hofmannii . J. Bacteriol. 92: 1304–1310. 1966.—A serologically active substance, extracted from sonically treated cells of Corynebacterium hofmannii with hot HCl, produced two precipitin lines by immunodiffusion tests with a hyperimmune homologous serum. Extracts of other species failed to precipitate with the hofmannii antiserum. The active fraction was eluted from a diethylaminoethyl cellulose column in the third adsorption peak at a linear concentration of 0.5 m KCl, and produced two precipitin lines which corresponded in identity to those formed by the acid extract. Separation of the antigens was achieved by rechromatography on a Sephadex G-200 column; the major antigen was designated A; the minor, B. The homogeneity and purity of each antigen was established by immunoelectrophoresis and, in addition, that of antigen A by disc electrophoresis. Biochemical analyses showed that both antigens were composed of a major protein component with polysaccharide and nucleic acid present in an approximate ratio of 17:3:1, respectively. Glutamic acid, aspartic acid, alanine, glycine, valine, and leucine were the main amino acids present. Antigen A contained 17% less protein and 3.5% less carbohydrate than antigen B. The principal sugars of antigen A were identified as arabinose and glucose. The molecular weight, estimated by gradient centrifugation, was 16,500 for antigen A and 21,000 for antigen B.
Precipitin
Immunodiffusion
Sephadex
Immunoelectrophoresis
Ouchterlony double immunodiffusion
Corynebacterium
Alanine
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Immunoelectrophoresis
Sephadex
Immunodiffusion
Ouchterlony double immunodiffusion
Radial immunodiffusion
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Immunoelectrophoresis
Taenia solium
Sephadex
Immunodiffusion
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ABSTRACT The purification procedures and characteristics of an equine infectious anemia (EIA) antigen reacting specifically in the immunodiffusion test with sera from EIA‐infected horses were investigated. Partially purified antigens were extracted from both infected horse leukocyte cultures and the horse spleen principally by dialysis against 0.01m acetate buffer, pH 5.0. These extracts were further purified by filtering through a Sephadex G‐100 column. The purified antigens had a common antigenic determinant and were heat‐labile and soluble at pH 5.0. The results of immunoelectrophoresis and gel filtration suggested that the spleen‐derived antigen was of a smaller molecule than the culture‐derived antigen. The partially purified antigens from both sources were comparatively used for the detection of precipitating antibody in experimentally infected horses and in horses in the field. The two antigens showed an identical reaction with each serum tested. The two partially purified antigens were considered to be useful for diagnosis of EIA.
Equine infectious anemia
Sephadex
Immunoelectrophoresis
Immunodiffusion
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