Mapping of monoclonal antibodies specific to P64k: a common antigen of several isolates of Neisseria meningitidis.
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.
División de Vacunas, Centro de Ingeniería Genética y Biotechnología, Habana, Cuba.
C
C
Nazábal
T
T
Carmenate
S
S
Cruz
S
S
González
R
R
Silva
A
A
Musacchio
M
M
Delgado
G
G
Chinea
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D008058:Dihydrolipoamide Dehydrogenase / Q000235:genetics / Q000276:immunology; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000276:immunology*; D017433:Protein Structure, Secondary
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; C112623:lpdA protein, Neisseria meningitidis; D008058:Dihydrolipoamide Dehydrogenase