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Identification and characterization of a novel heparin-binding peptide for promoting osteoblast adhesion and proliferation by screening an Escherichia coli cell surface display peptide library Hyoun-Ee Kim,a Hae-Won Kimb and Jun-Hyeog Jangc∗ Heparin/heparan sulfate (HS) plays a key role in cellular adhesion. In this study, we utilized a 12-mer random Escherichia coli cell surfacedisplay library to identify thesequence,whichbinds toheparin. Isolated insertanalysis revealedanovelheparin-binding peptide sequence, VRRSKHGARKDR, designated as HBP12. Our analysis of the sequence alignment of heparin-binding motifs known as the Cardin–Weintraub consensus (BBXB, where B is a basic residue) indicates that the HBP12 peptide sequence contains two consecutive heparin-binding motifs (i.e. RRSK and RKDR). SPR-based BIAcore technology demonstrated that the HBP12 peptide binds to heparin with high affinity (KD = 191 nM). The HBP12 peptide is found to bind the cell surface HS expressed by osteoblastic MC3T3 cells and promote HS-dependent cell adhesion. Moreover, the surface-immobilized HBP12 peptide on titanium substrates shows significant increases in the osteoblastic MC3T3-E1 cell adhesion and proliferation