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Directed evolution of novel binding proteins

  • US 6,979,538 B2
  • Filed: 02/14/2001
  • Issued: 12/27/2005
  • Est. Priority Date: 09/02/1988
  • Status: Expired due to Fees
First Claim
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1. A method of obtaining a nucleic acid encoding a proteinaceous binding domain that binds a predetermined target material comprising:

  • (a) expressing a plurality of different chimeric potential binding proteins, each said protein comprising (i) a potential binding domain which is an antibody variable domain and (ii) an outer surface transport signal for obtaining the display of the potential binding domain on the outer surface of a filamentous phase,(b) displaying said chimeric potential binding proteins on the outer surfaces of a population of filamentous phage, said population thereby collectively displaying a plurality of different potential binding domains which differ from each other in amino acid sequence, each phage including a nucleic acid construct comprising a nucleic acid sequence encoding one of said binding domains;

    (c) contacting said phage with the target material such that said potential binding domains and the target material may interact;

    (d) separating phage displaying a potential binding domain that binds the target material from phage that do not so bind, and(e) recovering at least one phage displaying on its outer surface a chimeric binding protein comprising a successful binding domain (SBD) which bound said target, said phage enclosing SBD-encoding nucleic acid, and amplifying said SBD-encoding nucleic acid in vivo or in vitro.

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