Author
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Schmerr, Mary Jo |
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Goodwin, Kathryn |
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Cutlip, Randall |
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Submitted to: International Immunology Congress
Publication Type: Abstract Only Publication Acceptance Date: 7/24/1995 Publication Date: N/A Citation: N/A Interpretive Summary: Technical Abstract: Transmissible spongiform encephalopathies of animals and humans are infectious diseases that cause degenerative disorders of the central nervous system. These diseases are caused by a post-translational modification of a cellular protein found primarily on the surface of neurons. This modified protein (PrPsc) aggregates into rod-shaped fibrils in the brains of infected animals and is resistant to proteases. The infected animals mount no observable immune response. Currently, there is a need for a new method to detect the presence of PrPsc that requires less material than conventional methods. We developed a method to measure immunocomplex formation using capillary electrophoresis with laser induced fluorescence for detection (CE-LIF). Two rabbit antisera which were made to two different peptides that define antigenic sites of PrPsc were affinity purified with the corresponding peptide and then labeled with fluorescein isothiocyanate. CE-LIF was used to detect immunocomplex formation with PrPsc and the labeled antibodies. Immunocomplexes were observed from samples prepared from infected sheep brain, but not from normal sheep brain. When the reaction mixtures were incubated overnight in the cold, more immunocomplex formation was observed than when the reaction mixtures were incubated for one-fourth to 1 hour at 25 degrees C. The respective peptide for each antibody reduced the amount of immunocomplex formation. The small sample size required and the automated equipment make this a potential diagnostic test for PrPsc. |
