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COMBINATORIAL PROTEIN LIBRARY SCREENING BY PERIPLASMIC EXPRESSION

  • US 20070099267A1
  • Filed: 08/22/2006
  • Published: 05/03/2007
  • Est. Priority Date: 10/27/2000
  • Status: Abandoned Application
First Claim
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1. A method of obtaining a bacterium comprising a nucleic acid sequence encoding a binding polypeptide having specific affinity for a target ligand comprising the steps of:

  • (a) providing a Gram negative bacterium comprising an inner and an outer membrane and a periplasm, said bacterium expressing a nucleic acid sequence encoding a candidate binding polypeptide, wherein the candidate binding polypeptide is exposed within the periplasm of said bacterium;

    (b) contacting the bacterium with a labeled ligand under conditions wherein the labeled ligand is capable of contacting the binding polypeptide; and

    (c) selecting said bacterium based on the presence of said labeled ligand bound to said candidate binding polypeptide.

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