Production and characterization of strain-specific monoclonal antibodies against outer membrane components of Azospirillum brasilense Sp245

Several hybridoma cell lines producing murine monoclonal antibodies (MAbs) directed against outer membrane components of the Gram-negative rhizosphere bacterium Azospirillum brasilense Sp245 have been established and characterized. Whole bacterial cells were used as immunogens. Among the clones obtained, 14 hybridoma cell lines were selected for further characterization. Eight MAbs were strain-specific and 6 MAbs showed cross- reactivity with a closely related strain Azospirillum brasilense Sp246. According to the biochemical characterization of the antigenic determinants, MAbs were classified into four groups. The corresponding antigens were lipopolysaccharides (class 1) and an outer membrane protein (class 4), which is common to Azospirillum brasilense Sp245 and Azospirillum brasilense Sp246 as well as two outer membrane proteins (class 2 and class 3) that are characteristic for Azospirillum brasilense Sp245. The number of antigens per cell varied from 4000 (class 1) to 100 (class 4). In each class high affinity MAbs were identified, which made a sensitive direct quantification of Azospirillum brasilense Sp245 possible.