Bioavailability of immobilized epidermal growth factor: covalent versus noncovalent grafting

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1116/1.4978871
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Name affiliation
  1. National Research Council of Canada. Human Health Therapeutics
FormatText, Article
Journal titleBiointerphases
ISSN1934-8630
1559-4106
Volume12
Issue1
Article number010501
Abstract
In an effort to rationalize and optimize an antiapoptotic coating combining chondroitin sulfate (CS) and epidermal growth factor (EGF) for vascular applications, the authors here report the comparison of two grafting strategies aiming to display EGF in an oriented fashion on CS. For that purpose, the authors produced, purified, and characterized a chimeric protein corresponding to EGF that was N-terminally fused to a cysteine and a coil peptide. The chimera was covalently immobilized via its free thiol group or captured via coiled–coil interactions at the surface of a biosensor or on a chondroitin sulfate coating in multiwell plates, mimicking the coating that was previously developed by them for stent-graft surfaces. The interactions of grafted EGF with the soluble domain of its receptor or the impact of grafted EGF upon vascular smooth muscle survival in proapoptotic conditions indicated that the coiled–coil based tethering was the best approach to display EGF. These results, combined to direct enzyme-linked immunosorbent assay measurements, indicated that the coiled–coil tethering approach allowed increasing the amount of bioavailable EGF when compared to covalent coupling, rather than the total amount of grafted EGF, while using much lower concentrations of tagged EGF during incubation.
Publication date
PublisherAIP Publishing
LanguageEnglish
Peer reviewedYes
NPARC number23001806
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Record identifier258b93df-81b6-4735-a94a-c95251c893b7
Record created2017-04-07
Record modified2020-03-16
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