A multifunctional fusion peptide for tethering to hydroxyapatite and selective capture of bone morphogenetic protein from extracellular milieu.

Abstract

The present study sought to design a multi-functional fusion peptide with hydroxyapatite (HA) binding domain (HABD) and heparin binding domain (HBD). The 74 amino acid fusion peptide contained N-terminus of the fibrinogen β chain (β 15-66), double G4S-linker and 12 residues with HA affinity. This construct was designed, synthesized and cloned into pET21a(+) vector and expressed in E. coli. HABD facilitated purification of the fusion peptide by HA affinity chromatography. Kinetic peptide binding and release on HA scaffold showed sustained release of peptide for up to 16 days. Competitive ELISA and intrinsic fluorescence assays were applied to determine HBD affinity to bone morphogenetic protein-2 (BMP-2). The disassociation rate constant (K_d ) for HBD and rhBMP-2 was approximately 9.2-12 nM. The fusion peptide developed in the present study, allowed for streamlined purification on HA affinity chromatography, as well as sustained release from HA scaffold, attributed to its HABD. HBD mediated binding to BMP-2, which may be potentially useful for bone repair. Additional studies, including in vivo investigation will be required to assess the efficacy of the fusion peptide in bone tissue engineering.