Development, Characterization and Future Prospects of Monoclonal Antibodies Against Spores of Glugea atherinae (Protozoa‐Microsporidia‐Fish Parasites)

Abstract

Monoclonal antibodies against spores of Glugea atherinae were obtained after lymphocytic hybridization made from immunized mouse splenocytes. Screening using an indirect enzyme linked immunosorbent assay (ELISA), revealed seven monoclonal antibodies with an intense but variable reaction with the spores of fish microsporidia, and a moderate reaction with those of an insect microsporidium (Nosema sp.). The reaction was weaker with spores of Encephalitozoon intestinalis found in HIV' patients. FITC and Dot Blot confirmed the majority of these results. After biotinylation of the seven antibodies, inhibition tests allowed the localization of two different recognition domains on the spores of Glugea atherinae. The multiple antigenic determinants and their probable polysaccharide nature seem to be in accord with the class IgM of the antibodies produced. This work confirms the potential of these antibodies for microsporidian taxonomy and diagnosis, especially the use of Mabs 12F9 and 12H5 for detection of spores in stools of HIV' patients.

Production and characterization of monoclonal antibodies against Encephalitozoon intestinalis and Encephalitozoon sp. spores and their developmental stages

“…ELISA 96 microtitter plates (Nunc-Immuno Plate PolySorp™, Darmstadt, Germany) were coated with E. intestinalis soluble antigen (0.8 μg/ml) as described previously [ 4 ]. Hybridomas with an optical density (OD) greater or equal to 0.5 were selected following the criteria of Pomport-Castillon et al [ 34 ].…”