title: Directed evolution of a protein: selection of potent neutrophil elastase inhibitors displayed on M13 fusion phage.
abstract: Inhibitors of human neutrophil elastase were engineered by designing and producing a library of phage-displayed protease inhibitory domains derived from wild-type bovine pancreatic trypsin inhibitor and fractionating the library for binding to the target protease. The affinity of one of the engineered variants for human neutrophil elastase (Kd = 1.0 pM) is 3.6 x 10(6)-fold higher than that of the parental protein and exceeds the highest affinity reported for any reversible human neutrophil elastase inhibitor by 50-fold. Thus the display phage method has allowed us to obtain protein derivatives that exhibit greatly increased affinity for a predetermined target. The technology can be applied to design high-affinity proteins for a wide variety of target molecules.
authors.affiliation: Protein Engineering Corporation, Cambridge, MA 02138.
authors.forename: B L
authors.initials: BL
authors.lastname: Roberts
authors.forename: W
authors.initials: W
authors.lastname: Markland
authors.forename: A C
authors.initials: AC
authors.lastname: Ley
authors.forename: R B
authors.initials: RB
authors.lastname: Kent
authors.forename: D W
authors.initials: DW
authors.lastname: White
authors.forename: S K
authors.initials: SK
authors.lastname: Guterman
authors.forename: R C
authors.initials: RC
authors.lastname: Ladner
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003090:Coliphages / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D006801:Humans; D007700:Kinetics; D019272:Leukocyte Elastase; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010196:Pancreatic Elastase / Q000037:antagonists & inhibitors*; D011480:Protease Inhibitors / Q000302:isolation & purification / Q000494:pharmacology*; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D009838:Oligodeoxyribonucleotides; D011480:Protease Inhibitors; D011994:Recombinant Proteins; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase