Preparation and purification of monoclonal antibodies for Aspergillus fumigatus.
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概要
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Monoclonal antibodies were raised against <I>Aspergillus fumigatus</I>, a fungus which is an etiological agent of aspergillosis. Male SPF BALB/c mice were immunized subcutaneously with <I>A. fumigatus</I> culture-filtrate or mycelial antigen. Their splenocytes were hybridized with mouse myeloma cells, P3 X 63 Ag 8.653 The hybridoma supernatants were screened by enzyme-linked immunosorbent assay (ELISA), using <I>A. fumigatus</I> culture-filtrate antigen. Of the 123 hybridoma clones, 27 cell lines secreting antibodies positive for <I>A. fumigatus</I> were obtained. Two of these cell lines were further grown to larger volumes. The monoclonal antibodies obtained were immunoglobulin-M (IgM) antibodies, which were then purified by ammonium sulphate precipitation and Mono Q anion exchange chromatography. The purified monoclonal antibodies showed reactivity against the protein constituent of the culture-filtrate antigen.
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