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Monovalent phage display

  • US 5,821,047 A
  • Filed: 06/05/1995
  • Issued: 10/13/1998
  • Est. Priority Date: 12/03/1990
  • Status: Expired due to Term
First Claim
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1. A method for selecting novel binding polypeptides comprising:

  • (a) constructing a replicable expression vector comprising a transcription regulatory element operably linked to a gene fusion encoding a fusion protein wherein the gene fusion comprises a first gene encoding a polylpeptide, and a second gene encoding at least a portion of a phage coat protein;

    (b) mutating the vector at one or more selected positions within the first gene thereby forming a family of related plasmids;

    (c) transforming suitable host cells with the plasmids;

    (d) infecting the transformed host cells with an amount of a helper phage having a gene encoding the phage coat protein sufficient to produce recombinant phagemid particles wherein no more than a minor amount of phagemid particles display more than one copy of the fusion protein on the surface of the particle;

    (e) culturing the transformed infected host cells under conditions suitable for forming recombinant phagemid particles containing at least a portion of the plasmid and capable of transforming the host cells, the conditions adjusted so that no more than a minor amount of phagemid particles display one or more copies of the fusion protein on the surface of the particle;

    (f) contacting the phagemid particles with a target molecule so that at least a portion of the phagemid particles bind to the target molecule; and

    (g) separating the phagemid particles that bind from those that do not.

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