title: Pathogen-free screening of bacteria-specific hybridomas for selecting high-quality monoclonal antibodies against pathogen bacteria as illustrated for Legionella pneumophila.
abstract: Antibodies are potent biological tools increasingly used as detection, diagnostic and therapeutic reagents. Many technological advances have optimized and facilitated production and screening of monoclonal antibodies. We report here an original method to screen for antibodies targeting biosafety level 2 or 3 pathogens without the fastidious handling inherent to pathogen use. A double ELISA screening was performed using as coated antigen transformed Escherichia coli expressing at its surface a protein specific to the pathogenic bacteria versus control untransformed E. coli. This method was applied to Legionella, using the surface-exposed Mip protein (macrophage infectivity potentiator). This screening proved to be an excellent means of selecting mAbs that bind Legionella pneumophila 1 surface-exposed Mip protein. This method also appears more biologically relevant than screening using the recombinant Mip protein alone and less tedious than a test performed directly on Legionella bacteria. We obtained 21 mAbs that bind strongly to L. pneumophila serogroups 1 to 13, and we validated their use in a rapid ELISA (performed in 4.5 h) and an immunochromatographic test (20 min).
authors: CEA, iBiTec-S, Service de Pharmacologie et d'Immunoanalyse, CEA Saclay, 91191 Gif-sur-Yvette, France. cecile.feraudet-tarisse@cea.fr
authors: Cécile
authors: C
authors: Féraudet-Tarisse
authors: Marja-Liisa
authors: ML
authors: Vaisanen-Tunkelrott
authors: Karine
authors: K
authors: Moreau
authors: Patricia
authors: P
authors: Lamourette
authors: Christophe
authors: C
authors: Créminon
authors: Hervé
authors: H
authors: Volland
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology* / Q000302:isolation & purification; D058846:Antibodies, Monoclonal, Murine-Derived / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006825:Hybridomas; D016952:Legionella pneumophila / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity; D051379:Mice; D019696:Peptidylprolyl Isomerase / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D015203:Reproducibility of Results
chemical_list: D000907:Antibodies, Bacterial; D058846:Antibodies, Monoclonal, Murine-Derived; D001426:Bacterial Proteins; D011994:Recombinant Proteins; C512922:Mip protein, Legionella pneumophila; D019696:Peptidylprolyl Isomerase