Immunological fingerprint of 4CMenB recombinant antigens via protein microarray

In this study, we describe the development and use of an ad hoc protein microarray to study the immune response induced by the three major 4CMenB antigenic components (fHbp, NHBA and NadA) in individual sera from vaccinated infants, adolescents and adults. 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 a 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. Sera were profiled for anti-Human IgG specific analysis of reactivity to the spotted proteins.