To investigate whether specific peptide sequences could induce virion transport across the intestinal barrier, we used phage display to both identify signalling peptides capable of inducing trans-intestinal transport and also provide a suitable model of virion translocation. We utilised simple, single-round high input in vivo biopanning protocol using a 7-mer random amino acid phage display library. Phage were applied by gavage and translocation across the intestinal barrier assessed by phage recovery from the spleen 2h later. Following isolation, a number of phage were sequenced and several homologies with HIV gp120 were identified. Immunocytochemical analysis of phage translocation across the intestinal barrier by a phage bearing the peptide YPRLLTP demonstrated that phage were actively transported along specific channels. It is concluded that utilisation of in vivo phage display (IVPD) has provided evidence for a specific peptide-guided transport of undegraded cargo across the intestinal barrier, modelled by M13 phage.