A hexapeptide phage library was used to select peptides with affinity for the tumor-associated TAG72 antigen. Twenty-one phage clones were selected after the third round of biopanning. Three phage clones with the same DNA insert of ARTLRF were found to bind more strongly to the TAG72 antigen than other phage clones and the wild-type phage. A synthetic decapeptide GAARTLRFGA with two conjunctive amino acid residues of the phage coat protein III on each side of the selected peptide was found to bind more strongly to the TAG72 antigen than to other antigens such as the mouse metallothionein. Furthermore, immunohistochemical studies revealed that this peptide displayed preferential binding to colonic adenocarcinomatous cells expressing the TAG72 antigen. Therefore, this anti-TAG72 peptide may be useful in serving as the starting point with regard to further designing peptidomimetics as potential pharmaceuticals.