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Monoclonal antibodies to yeast catalase T
Authors:Günther R Adolf  Engelbert Hartter  Helmut Ruis  Peter Swetly
Institution:1. Ernst-Boehringer-Institut für Arzneimittelforschung, A-1121 Vienna, Austria;2. Institut für Allgemeine Biochemie der Universität Wien, A-1090 Vienna, Austria.;3. Ludwig Boltzmann-Forschungsstelle für Biochemie, A-1090 Vienna, Austria.
Abstract:Hybridoma cell lines secreting antibodies directed against Saccharomyces cerevisiae catalase T were constructed by fusing spleen cells of mice immunized with catalase T with P3×63Ag8 mouse myeloma cells. Culture supernatants were assayed for specific antibodies by incubation with 35S-labelled yeast extracts, adsorption of the immune complexes to Protein A — carrying Staphylococus aureus cells and analysis of the adsorbed yeast proteins by sodium dodecylsulfate gel electrophoresis. Two hybrid clones were isolated mediating adsorption of a protein with electrophoretic mobility of catalase T; one of them, showing considerably higher activity, was characterized further. Anti-bodies produced by this clone belong to the IgG class of immuniglobulins; they can be used for immunoadsorption, but not for direct immunoprecipitation and recognize authentic catalase T as well as catalase T apoprotein.
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