Cellular libraries of peptide sequences (CLiPS) and methods of using the same

摘要

The present invention provides compositions including peptide display scaffolds that present at least one candidate peptide and at least one detectable moiety in at least one of the N-terminal and C-terminal candidate peptide presenting domains that when expressed in a cell are accessible at a surface of the cell outermembrane. In addition, the present invention also provides kits and methods for screening a library of cells presenting the candidate peptides in peptide display scaffolds to identify a ligand for an enzyme.

主权项

1. A peptide display scaffold, comprising a fusion protein comprising:
a D1 moiety positioned N-terminal to a C1 moiety in the fusion protein, which C1 moiety is positioned N-terminal to a TM moiety in the fusion protein, which TM moiety is positioned N-terminal to a D2 moiety in the fusion protein; or a D2 moiety positioned N-terminal to a TM moiety in the fusion protein, which TM moiety is positioned N-terminal to a C1 moiety in the fusion protein, which C1 moiety is positioned N-terminal to a D1 moiety in the fusion protein; wherein the TM moiety is a circularly permuted bacterial outer membrane protein; the C1 moiety comprises a candidate peptide; and the D1 and D2 moieties are first and second detectable moieties, wherein the D1 and D2 moieties are not the same, wherein the D1 moiety and the D2 moiety are each a heterologous peptide or polypeptide relative to the circularly permuted bacterial outer membrane protein, and wherein, when the fusion protein is expressed in a cell comprising a cell outer membrane, the D1, C1 and D2 moieties are exposed at an extracellular surface of the cell outer membrane and one or both of the N- and C-termini of the fusion protein are exposed at the extracellular surface of the cell outer membrane.