title: Flow-based enzymatic ligation by sortase A.
abstract: Sortase-mediated ligation (sortagging) is a versatile, powerful strategy for protein modification. Because the sortase reaction reaches equilibrium, a large excess of polyglycine nucleophile is often employed to drive the reaction forward and suppress sortase-mediated side reactions. A flow-based sortagging platform employing immobilized sortase A within a microreactor was developed that permits efficient sortagging at low nucleophile concentrations. The platform was tested with several reaction partners and used to generate a protein bioconjugate inaccessible by solution-phase batch sortagging.
authors.affiliation: Department of Chemistry, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 (USA).
authors.forename: Rocco L
authors.initials: RL
authors.lastname: Policarpo
authors.forename: Hansol
authors.initials: H
authors.lastname: Kang
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Liao
authors.forename: Amy E
authors.initials: AE
authors.lastname: Rabideau
authors.forename: Mark D
authors.initials: MD
authors.lastname: Simon
authors.forename: Bradley L
authors.initials: BL
authors.lastname: Pentelute
mesh_terms: D019881:Aminoacyltransferases / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D008958:Models, Molecular; D015394:Molecular Structure; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004800:Enzymes, Immobilized; D011506:Proteins; D019881:Aminoacyltransferases; C120542:sortase A; D003546:Cysteine Endopeptidases
keywords: flow chemistry; immobilization; microreactor; protein modification; sortase A