Construction and Selection of Affilin® Phage Display Libraries
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Affilin® molecules represent a new class of so-called scaffold proteins. The concept of scaffold proteins is to use stable and versatile protein structures which can be endowed with de novo binding properties and specificities by introducing mutations in surface exposed amino acid residues. Complex variations and combinations are generated by genetic methods of randomization resulting in large cDNA libraries. The selection for candidates binding to a desired target can be executed by display methods, especially the very robust and flexible phage display. Here, we describe the construction of ubiquitin based Affilin® phage display libraries and their use in biopanning experiments for the identification of novel protein ligands.
Key wordsAffilin® Library construction Phagemid TAT phage display Biopanning Selection Maturation
We thank Anja Kunert for early work on the protocols, Ulrich Haupts for helpful discussions, and Erik Fiedler for chromatographic purification of the phage libraries.
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