In situ regeneration of bioactive coatings enabled by an evolved Staphylococcus aureus sortase A.
Surface immobilization of bioactive molecules is a central paradigm in the design of implantable devices and biosensors with improved clinical performance capabilities. However, in vivo degradation or denaturation of surface constituents often limits the long-term performance of bioactive films. Here we demonstrate the capacity to repeatedly regenerate a covalently immobilized monomolecular thin film of bioactive molecules through a two-step stripping and recharging cycle. Reversible transpeptidation by a laboratory evolved Staphylococcus aureus sortase A (eSrtA) enabled the rapid immobilization of an anti-thrombogenic film in the presence of whole blood and permitted multiple cycles of film regeneration in vitro that preserved its biological activity. Moreover, eSrtA transpeptidation facilitated surface re-engineering of medical devices in situ after in vivo implantation through removal and restoration film constituents. These studies establish a rapid, orthogonal and reversible biochemical scheme to regenerate selective molecular constituents with the potential to extend the lifetime of bioactive films.
|Year of Publication||
2016 Apr 13
|PubMed Central ID||
R01 HL056819 / HL / NHLBI NIH HHS / United States
R35GM118062 / GM / NIGMS NIH HHS / United States
R01HL056819 / HL / NHLBI NIH HHS / United States
R01EB022376 / EB / NIBIB NIH HHS / United States
R35 GM118062 / GM / NIGMS NIH HHS / United States
Howard Hughes Medical Institute / United States
R01 EB022376 / EB / NIBIB NIH HHS / United States