title: In situ regeneration of bioactive coatings enabled by an evolved Staphylococcus aureus sortase A.
abstract: 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.
authors.affiliation: Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
authors.forename: Hyun Ok
authors.initials: HO
authors.lastname: Ham
authors.affiliation: Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.|Coulter Department of Biomedical Engineering, Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Qu
authors.affiliation: Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
authors.forename: Carolyn A
authors.initials: CA
authors.lastname: Haller
authors.affiliation: Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
authors.forename: Brent M
authors.initials: BM
authors.lastname: Dorr
authors.affiliation: Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
authors.forename: Erbin
authors.initials: E
authors.lastname: Dai
authors.affiliation: Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
authors.forename: Wookhyun
authors.initials: W
authors.lastname: Kim
authors.affiliation: Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
authors.forename: David R
authors.initials: DR
authors.lastname: Liu
authors.affiliation: Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.|Coulter Department of Biomedical Engineering, Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, Georgia 30332, USA.|Wyss Institute of Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA.
authors.forename: Elliot L
authors.initials: EL
authors.lastname: Chaikof
mesh_terms: D019881:Aminoacyltransferases / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D055162:Biocatalysis / Q000187:drug effects; D002406:Catheterization, Peripheral; D020099:Coated Materials, Biocompatible / Q000494:pharmacology*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D008810:Mice, Inbred C57BL; D017208:Rats, Wistar; D013211:Staphylococcus aureus / Q000201:enzymology*; D013499:Surface Properties
chemical_list: D001426:Bacterial Proteins; D020099:Coated Materials, Biocompatible; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases