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Hsingchen Chen - One of the best experts on this subject based on the ideXlab platform.

Kunshang Chen - One of the best experts on this subject based on the ideXlab platform.

Turid M Pedersen - One of the best experts on this subject based on the ideXlab platform.

  • detection of biotinylated proteins in Crossed Immunoelectrophoresis gels studies on platelet membrane receptors and microparticles
    Electrophoresis, 1995
    Co-Authors: Nils Olav Solum, Pal Andre Holme, Turid M Pedersen
    Abstract:

    Biotinylation can be used as an alternative for surface labeling of cell membrane proteins. The use of the water soluble N-hydroxysulfosuccinimide (NHSS)-biotin or the more lipophilic N-hydroxysuccinimide (NHS)-biotin reagent has been investigated in the present study labeling two central receptor complexes on the platelet surface, i.e. the glycoprotein (GP) Ib–IX and the GP IIb–IIIa complexes involved in platelet adhesion and aggregation. Lack of labeling of the intracellularly located albumin was used as a negative control. The labeling has been studied using Crossed Immunoelectrophoresis in the PhastSystem format after extraction of the labeled cells in Triton X-100, and it is shown that, using enzyme-conjugated avidin and chromogenic substrates, the biotinylated proteins can be visualized directly in the dried electrophoresis gel without the need for a transfer to a blotting membrane as is used after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Suitable conditions for biotinylation and for visualization in the Crossed Immunoelectrophoresis gels are described. Further, surface-biotinylation of platelets was used to observe shedding of microparticles as a consequence of formation of the complement membrane attack complex. For this purpose the formation and composition of the biotinylated microparticles were observed by flow cytometry and Crossed Immunoelectrophoresis.

K K Lee - One of the best experts on this subject based on the ideXlab platform.

N Hoiby - One of the best experts on this subject based on the ideXlab platform.

  • antibodies against pseudomonas aeruginosa in serum from normal persons and patients colonized with mucoid or non mucoid pseudomonas aeruginosa results obtained by Crossed Immunoelectrophoresis
    Acta Pathologica Microbiologica Scandinavica Section C Immunology, 2009
    Co-Authors: N Hoiby
    Abstract:

    Serum from 154 normal persons was investigated for precipitating antibodies against P. aeruginosa by Crossed Immunoelectrophoresis, using a polyvalent P. aeruginosa standard antigen. Ninety-four per cent of the sera contained no demonstrable precipitins, and 6 per cent contained precipitins against only one P. aeruginosa antigen. Thirty-four per cent of sera from 122 patients colonized with P. aeruginosa, in various anatomical regions contained precipitins against 2 to 41 P. aeruginosa antigens. Serum from 14 patients harbouring mucoid strains of P. aeruginsa contained precipitins against these bacteria, and the average number of precipitins was significantly higher than the average number of precipitins in serum from patients harbouring non-mucoid strains. The largest number of precipitins was found in serum from adult patients chronically colonized with mucoid P. aeruginosa in the lower respiratory tract, and from adult patients with P. aeruginosa bacteraemia originating from suppurative urinary tract infections cavised by mucoid strains. Serum from patients with P. aeruginosa cystitis or from patients with P. aeruginosa colonization of the middle ear or with wound infection, contained only few or no precipitins.

  • antibodies against pseudomonas aeruginosa in serum from normal persons and patients colonized with mucoid or non mucoid pseudomonas aeruginosa results obtained by Crossed Immunoelectrophoresis
    Acta Pathologica Microbiologica Scandinavica Section C Immunology, 2009
    Co-Authors: N Hoiby
    Abstract:

    Serum from 154 normal persons was investigated for precipitating antibodies against P. aeruginosa by Crossed Immunoelectrophoresis, using a polyvalent P. aeruginosa standard antigen. Ninety-four per cent of the sera contained no demonstrable precipitins, and 6% contained precipitins against only one P. aeruginosa antigen. Thirty-four per cent of sera from 122 patients colonized with P. aeruginosa in various anatomical regions contained precipitins against 2 to 41 P. aeruginosa antigens. Serum from 14 patients harbouring mucoid strains of P. aeruginsa contained precipitants against these bacteria, and the average number of precipitins was significantly higher than the average number of precipitins in serum from patients harbouring non-mucoid strains. The largest number of precipitins was found in serum from adult patients chronically colonized with mucoid P. aeruginosa in the lower respiratory tract, and from adult patients with P. aeruginosa bacteremia originating from suppurative urinary tract infections caused by mucoid strains. Serum from patients with P. aeruginosa cystitis or from patients with P. aeruginosa colonization of the middle ear or with wound infection, contained only few or no precipitins.

  • precipitating antibodies against pseudomonas aeruginosa in sputum from patients with cystic fibrosis specificities and titres determined by means of Crossed Immunoelectrophoresis with intermediate gel
    Acta Pathologica Microbiologica Scandinavica Section C Immunology, 2009
    Co-Authors: P. O. Schiøtz, N Hoiby
    Abstract:

    By means of Crossed Immunoelectrophoresis with intermediate gel, paired 24-h samples of sputum and sera from 19 patients with cystic fibrosis have been examined for the occurrence of precipitins against Pseudomonas aeruginosa. Saliva from 9 of the patients was subject to similar analyses. A total of 50 precipitins was found in fifteen of the sputa and 296 precipitins were found in 18 of the sera. Fourteen of the patients had precipitins in sputum as well as in serum; one patient had a precipitin in sputum, but none in serum; and 4 had precipitins in serum, but not in sputum. All the precipitins in sputa could be identified and quanti-tated in relation to the reference system and, in this way, 16 different precipitins were found in the sputa. If present in sputum, titres of these precipitins were nearly always found to be lower in sputum than those of the corresponding precipitin in serum from one and the same patient. One of the precipitins was, however, found in 8 of the sputa, but not in the corresponding sera. There was a positive correlation between the number of precipitins per sputum and between titre of the strongest precipitins and number of precipitins per serum. Pseudomonas antigens wefe demonstrated in 3 of the sputa. Saliva from the patients contained rarely precipitins.