Patent ID: 8759087
Filing Date: 2014-06-24
Classification: C12N

Abstract:
1. A method for recovering a nucleic acid molecule encoding a binding member of a complex internalized in a target cell, comprising the following steps of: (a) contacting a target cell with a diverse collection of bacteriophage particles, wherein each or substantially all of said bacteriophage particles display a binding member on its surface, wherein said binding member is displayed as a non-fusion (poly)peptide with a phage coat protein of said bacteriophage particle and wherein each or substantially all of said bacteriophage particles comprise a nucleic acid molecule encoding the displayed binding member, wherein said display as a non-fusion (poly)peptide is characterized by a disulfide bond between the phage coat protein and the binding member, (b) allowing for binding of the binding member displayed on the bacteriophage particle to its target, thereby allowing for the formation of at least one complex, each of said complexes comprising a bacteriophage particle with its displayed binding member and its target, (c) culturing the target cell under conditions that allow internalization of at least one of said complexes into the cell, (d) eluting the bacteriophage particles displaying a binding member that are not internalized under conditions that substantially no cell lysis occurs, wherein the eluting is carried out under reducing conditions such that said disulfide bond is cleaved, (e) lysing the target cell comprising the internalized complexes, and (f) recovering from the lysed cell the nucleic acid molecule encoding a binding member from at least one of the internalized complexes.