title: Flow cytometric sorting of bacterial surface-displayed libraries.
abstract: The protocols herein detail methods for isolating binding peptides from a combinatorial library displayed on the surface of bacterial cells. These methods are appropriate for a variety of display scaffolds and a large range of library sizes, up to approximately 5 x 10(9) or more. Instructions have been provided for isolating peptides that bind to both proteins and non-protein targets, such as whole cells or inorganic particles. Qualitative analysis by flow cytometry can be exploited for bacterial libraries to characterize a displayed peptide's binding properties with a target of interest, and sorting conditions can be tuned to maximize binding affinity.
authors.affiliation: University of California, Santa Barbara, California, USA.
authors.forename: Sophia
authors.initials: S
authors.lastname: Kenrick
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Rice
authors.forename: Patrick
authors.initials: P
authors.lastname: Daugherty
mesh_terms: D001419:Bacteria / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry; D001426:Bacterial Proteins / Q000032:analysis* / Q000737:chemistry; D001665:Binding Sites; D020650:Combinatorial Chemistry Techniques / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry; D005434:Flow Cytometry / Q000379:methods*; D006867:Hydrolases / Q000737:chemistry; D010455:Peptides / Q000032:analysis / Q000737:chemistry
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D010455:Peptides; C482905:OmpX protein, E coli; C078167:OMPA outer membrane proteins; D006867:Hydrolases