This EGFR specific binding protein (Z-domain of protein A) was investigated by an α-helix shuffling strategy. The primary scaffold protein was from a naive combinatorial library of the three-helix bundle Z domain derived from staphylococcal protein A. A hierarchical library was constructed through selective re-randomization of six amino acid positions in one of the two α-helices of the domain, making up the Taq DNA polymerase binding surface. After selections using monovalent phage display technology, second generation variants were identified having affinities (KD=25 nM ) for EGFR as determined by biosensor technology. It's potential to be used in diagnostic, research and therapeutic applications.
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