Mapping of monoclonal antibodies specific to P64k: A common antigen of several isolates of Neisseria meningitidis
Consuelo Nazábal, Tania Carmenate, S. A. B. da Cruz, Silvia A. González, R Silva, Alexis Musacchio, Maité Delgado, Glay Chinea
Abstract
Consuelo Nazábal, Tania Carmenate, S. A. B. da Cruz, Silvia A. González, R Silva, Alexis Musacchio, Maité Delgado, Glay Chinea
Abstract
P64k is a minor outer membrane protein from Neisseria meningitidis. This protein has been produced at high levels in Escherichia coli. We generated a group of monoclonal antibodies (mAbs) against recombinant P64k, which recognise four non-overlapping epitopes, as shown using competition assays with biotinylated mAbs. The P64k sequences involved in mAbs binding were mapped with synthetic overlapping peptides derived from the P64k protein, and located in the previously determined three-dimensional structure of the protein. These antibodies were also characterised by whole-cell ELISA and bactericidal tests against N. meningitidis. Only two of the recognised epitopes were exposed on the bacterial surface, and none of the mAbs showed bactericidal activity. The relationship between these results and the structural data on the epitopes bound by the mAbs is discussed.
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P64k is a minor outer membrane protein from Neisseria meningitidis. This protein has been produced at high levels in Escherichia coli. We generated a group of monoclonal antibodies (mAbs) against recombinant P64k, which recognise four non-overlapping epitopes, as shown using competition assays with biotinylated mAbs. The P64k sequences involved in mAbs binding were mapped with synthetic overlapping peptides derived from the P64k protein, and located in the previously determined three-dimensional structure of the protein. These antibodies were also characterised by whole-cell ELISA and bactericidal tests against N. meningitidis. Only two of the recognised epitopes were exposed on the bacterial surface, and none of the mAbs showed bactericidal activity. The relationship between these results and the structural data on the epitopes bound by the mAbs is discussed.
Key concepts: Epitope, Neisseria meningitidis, Monoclonal antibody, Epitope mapping, Microbiology, Neisseria, Bacterial outer membrane, Biology