Epitope mapping of a monoclonal antibody directed against Neisserial Heparin Binding Antigen by peptide microarray

The aim of the study was to determine the epitope targeted by 31E10/E7 mouse monoclonal antibody (mAb) and the cross-reactivity to linear peptide epitopes of 10 different Neisserial Heparin Binding Antigen (NHBA) variants. mAb 31E10/E7 was diluted at 1:200 and incubated on a custom PepStar Peptide Microarray platform printed with 560 different peptides in three independent experiments. A panel of 560 peptides (15-mer overlapping peptides with an offset of 4 amino acids) covering the entire sequence of ten different NHBA variants (p1, p2, p3, p5, p10, p17, p18, p20, p21 and p29) were printed in triplicate on a custom PepStar Peptide Microarray. Process-control peptide (GGSGGGSDYKDDDDK) and mouse IgG were distributed in each array copy as controls. mAb 31E10/E7 was profiled for anti-mouse IgG specific analysis of reactivity to the peptide epitopes