The aim of this study was to investigate whether immunization with the sugar binding domain of PapGII (PapGII196) was able to protect chickens against avian pathogenic Escherichia coli. PapGII196 was expressed, purified by Ni-NTA column chromatography and shown to retain its biological activity, as demonstrated by binding to its receptor, globoside. PapGII196 was tested as a vaccine in specific pathogen free broilers and also by vaccinating breeders and assessing protection in their offspring, and using aerosol exposure or air sac inoculation for challenge. Notwithstanding a strong anti-PapGII196 serum IgG response in vaccinated birds in all experiments and inhibition of haemagglutination by serum from PapGII196-vaccinated birds, chickens were not protected against avian pathogenic E. coli. These findings suggest that PapGII may not be a useful candidate for inclusion in vaccines against avian pathogenic E. coli.