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

  • US 9,234,847 B2
  • Filed: 12/23/2009
  • Issued: 01/12/2016
  • Est. Priority Date: 08/31/2005
  • Status: Active Grant
First Claim
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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;

    ora 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;

    whereinthe 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.

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