Bacteriophage surface display of an immunoglobulin-binding domain of Staphylococcus aureus protein A

Biotechnology (N Y). 1994 Feb;12(2):169-72. doi: 10.1038/nbt0294-169.


As a model system for the optimization of separation ligands by bacteriophage surface display, we have constructed a phage surface expression system for a single immunoglobulin-binding domain of Protein A of Staphylococcus aureus. Protein A domain B is genetically fused to the gpIII adsorption protein of the filamentous bacteriophage M13, and hence displayed on the phage surface. Phage displaying the Protein A domain are selectively retained on human IgG-sepharose. Retention is due to specific Protein A-IgG interactions, as demonstrated by competitive inhibition by soluble Protein A or polyclonal human IgG. Polyclonal goat IgG, which is known to bind less well to Protein A than does human IgG, inhibits phage adsorption less effectively. Phage expressing Protein A can be purified in a few rounds of selective adsorption from a vast excess of wild type phage. Diverse libraries constructed by mutagenesis of this construct will allow massive screening of mutant forms of Protein A for alterations in binding and elution properties. We anticipate that phage display will prove to be a widely-applicable method of identification and optimization of affinity ligands for separations.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Bacteriophage M13
  • Base Sequence
  • Binding Sites
  • Chromatography, Affinity
  • Cloning, Molecular / methods
  • Genes, Synthetic
  • Humans
  • Immunoglobulin G / metabolism*
  • Molecular Sequence Data
  • Oligodeoxyribonucleotides / chemical synthesis
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Staphylococcal Protein A / biosynthesis
  • Staphylococcal Protein A / isolation & purification
  • Staphylococcal Protein A / metabolism*
  • Staphylococcus aureus / genetics
  • Staphylococcus aureus / metabolism*


  • Immunoglobulin G
  • Oligodeoxyribonucleotides
  • Recombinant Proteins
  • Staphylococcal Protein A