Epitope mapping of human Fabs directed against 4CmenB protein components by protein microarray

The aim of the study was to determine the epitope targeted by a panel of Human Fabs. Fabs were diluted at 1:50 and incubated on a non-commercial Protein Microarray platform printed with fHbp, NHBA and NadA specific recombinant protein fragments and full length fHbp, NHBA and NadA of different variants. A panel of recombinant fragments of different length spanning the entire sequence of fHbp, NHBA and NadA plus different full length fHbp, NHBA and NadA variants were printed in 8 or 16 replicated on non-commercial Protein Microarray. All fragments were expressed in E. coli as either glutathione S-transferase-, His-tagged or TRX-fusions, purified from the cytoplasmic fraction as soluble forms. Human IgG (at 8 different concentration points) was distributed in each array copy as control. Fabs were profiled for anti-Human IgG specific analysis of reactivity to the spotted proteins.