Coiled-coil-mediated grafting of bioactive vascular endothelial growth factor

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Journal titleActa Biomaterialia
Pages68066813; # of pages: 8
Subjectchimeric protein; homodimer; vasculotropin; vasculotropin 165; article; assay; biological activity; cell survival; controlled study; human; human cell; immobilization; in vitro study; phenotype; priority journal; protein binding; surface plasmon resonance; Mammalia
AbstractChimeric growth factors may represent a powerful alternative to their natural counterparts for the functionalization of tissue-engineered scaffolds and applications in regenerative medicine. Their rational design should provide a simple, readily scalable production strategy while improving retention at the site of action. In that endeavor, we here report the synthesis of a chimeric protein corresponding to human vascular endothelial growth factor 165 being N-terminally fused to an E5 peptide tag (E5-VEGF). E5-VEGF was successfully expressed as a homodimer in mammalian cells. Following affinity purification, in vitro surface plasmon resonance biosensing and cell survival assays confirmed diffusible E5-VEGF ability to bind to its receptor ectodomains, while observed morphological phenotypes confirmed its anti-apoptotic features. Additional surface plasmon resonance assays highlighted that E5-VEGF could be specifically captured with high stability when interacting with covalently immobilized K5 peptide (a synthetic peptide designed to bind to the E5 moiety of chimeric hVEGF). This immobilization strategy was applied to glass substrates and chimeric hVEGF was shown to be maintained in a functionally active state following capture. Altogether, our data demonstrated that stable hVEGF capture can be performed via coiled-coil interactions without impacting hVEGF bioactivity, thus opening up the way to future applications in the field of tissue engineering and regenerative medicine. © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.
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AffiliationNational Research Council Canada (NRC-CNRC); Human Health Therapeutics (HHT-TSH)
Peer reviewedYes
NPARC number21269799
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Record identifier2562da32-8212-437f-b13a-7ddcf3d6e668
Record created2013-12-13
Record modified2016-05-09
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