To　screen　and　identify　the　novel　probe　markers　binding　hepatocellular　carcinoma　specifically　and　sensitively,　a　phage-displayed　12-mer　peptide　library　was　used　to　make　biopanning　with　the　modified　protocols　on　HepG2　cells.　After　four　rounds　of　panning,　the　consensus　sequences　were　obtained,　and　the　PC28,　a　phage　clone　with　most　specific　and　sensitive　binding　to　HepG2　cells,　was　identified　as　the　best　positive　clone.　The　peptide　probe　HCSP4　(sequence　SLDSTHTHAPWP)　was　synthesized　based　on　the　sequencing　result　of　PC28.　The　specificity　and　sensitivity　of　HCSP4　were　primarily　analyzed　using　immunofluorescence,　flow　cytometry,　and　other　methods.　The　results　show　that　HCSP4　can　bind　to　hepatocellular　carcinoma　cells　with　satisfactory　specificity　and　sensitivity.　It　may　be　a　promising　lead　candidate　for　molecular　imaging　and　targeted　drug　delivery　in　the　diagnosis　and　therapy　of　hepatocellular　carcinoma.　Copyright　(c)　2014　European　Peptide　Society　and　John　Wiley　&　Sons,　Ltd.