title: Microelectrode array biosensor for studying carbohydrate-mediated interactions.
abstract: Carbohydrate-mediated host-pathogen interactions are essential to bacterial and viral pathogenesis, and represent an attractive target for the development of antiadhesives to prevent infection. We present a versatile microelectrode array-based platform to investigate carbohydrate-mediated protein and bacterial binding, with the objective of developing a generalizable method for screening inhibitors of host-microbe interactions. Microelectrode arrays are well suited for interrogating biological binding events, including proteins and whole-cells, and are amenable to electrochemical derivitization, facilitating rapid deposition of biomolecules. In this study, we achieve microelectrode functionalization with carbohydrates via controlled polymerization of pyrrole to individual microelectrodes, followed by physisorption of neoglycoconjugates to the polypyrrole-coated electrodes. Bioactivity of the immobilized carbohydrates was confirmed with carbohydrate-binding proteins (lectins) detected by both fluorescent and electrochemical means. The platform's ability to analyze whole-cell binding was demonstrated using strains of Escherichia coli and Salmonella enterica, and the dose-dependent inhibition of S. enterica by a soluble carbohydrate antiadhesive.
authors.affiliation: Department of Bioengineering, University of Washington, Seattle, WA 98195, USA. jeffwc@uw.edu
authors.forename: Jeffrey W
authors.initials: JW
authors.lastname: Chamberlain
authors.forename: Karl
authors.initials: K
authors.lastname: Maurer
authors.forename: John
authors.initials: J
authors.lastname: Cooper
authors.forename: Wanda J
authors.initials: WJ
authors.lastname: Lyon
authors.forename: David L
authors.initials: DL
authors.lastname: Danley
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Ratner
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002241:Carbohydrates / Q000737:chemistry*; D002448:Cell Adhesion; D055664:Electrochemical Techniques; D004926:Escherichia coli / Q000737:chemistry; D054884:Host-Pathogen Interactions; D037102:Lectins / Q000737:chemistry; D008839:Microelectrodes; D011108:Polymers / Q000737:chemistry*; D011485:Protein Binding; D011506:Proteins / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D011758:Pyrroles / Q000737:chemistry*; D012276:Ricin / Q000302:isolation & purification; D019779:Salmonella enterica / Q000737:chemistry
chemical_list: D002241:Carbohydrates; D037102:Lectins; D011108:Polymers; D011506:Proteins; D011758:Pyrroles; C067635:polypyrrole; D012276:Ricin