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A multifunctional fusion peptide for tethering to hydroxyapatite and selective capture of bone morphogenetic protein from extracellular milieu
Author(s) -
Bahri Mina,
Hasannia Sadegh,
Dabirmanesh Bahareh,
Moshaverinia Alireza,
Zadeh Homayoun H.
Publication year - 2020
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.36915
Subject(s) - peptide , fusion protein , affinity chromatography , bone morphogenetic protein 2 , bone morphogenetic protein , biochemistry , biophysics , in vivo , linker , fusion , materials science , recombinant dna , microbiology and biotechnology , chemistry , in vitro , biology , enzyme , linguistics , philosophy , gene , computer science , operating system
Purpose The present study sought to design a multi‐functional fusion peptide with hydroxyapatite (HA) binding domain (HABD) and heparin binding domain (HBD). Methods 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 . Results 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. Conclusion 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.