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.
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