title: Isolation of a peptide ligand for affinity purification of factor VIII using phage display.
abstract: Polypeptides for use in affinity chromatography of factor VIII were identified using phage display technology. Phage libraries were designed to express polypeptide fusions containing five to seven residues flanked by two cysteines that form a disulfide bond. Individual bacteriophage were selected for the ability of these polypeptides to bind factor VIII, and then release the protein under mild elution conditions. Strong consensus sequences were observed that appear to be necessary for this reversible interaction. Chemically synthesized ligands identified by this screening were immobilized onto a chromatographic support and used for affinity purification of factor VIII from a complex feedstream. A chromatographic step was developed that provided a 10000-fold reduction in host cell proteins and DNA, while providing exceptional product recovery.
authors.affiliation: Wyeth BioPharma, 1 Burtt Road, Andover, MA 01810, USA. bkelley@wyeth.com
authors.forename: Brian D
authors.initials: BD
authors.lastname: Kelley
authors.forename: James
authors.initials: J
authors.lastname: Booth
authors.forename: Molly
authors.initials: M
authors.lastname: Tannatt
authors.forename: Qi-Long
authors.initials: QL
authors.lastname: Wub
authors.forename: Robert
authors.initials: R
authors.lastname: Ladner
authors.forename: Jinan
authors.initials: J
authors.lastname: Yuc
authors.forename: Daniel
authors.initials: D
authors.lastname: Potter
authors.forename: Arthur
authors.initials: A
authors.lastname: Ley
mesh_terms: D001435:Bacteriophages / Q000235:genetics*; D002846:Chromatography, Affinity / Q000379:methods*; D005169:Factor VIII / Q000235:genetics / Q000302:isolation & purification*; D010455:Peptides / Q000737:chemistry*
chemical_list: D010455:Peptides; D005169:Factor VIII