Genotype Phenotype Coupling pp 19-41 | Cite as
Affitins: Ribosome Display for Selection of Aho7c-Based Affinity Proteins
Protocol
First Online:
Abstract
Engineered protein scaffolds have made a tremendous contribution to the panel of affinity tools owing to their favorable biophysical properties that make them useful for many applications. In 2007, our group paved the way for using archaeal Sul7d proteins for the design of artificial affinity ligands, so-called Affitins. For many years, Sac7d and Sso7d have been used as molecular basis to obtain binders for various targets. Recently, we characterized their old gifted protein family and identified Aho7c, originating from Acidianus hospitalis, as the shortest member (60 amino-acids) with impressive stability (96.5 °C, pH 0–12). Here, we describe the construction of Aho7c combinatorial libraries and their use for selection of binders by ribosome display.
Key words
Ribosome display In vitro selection Sul7d Aho7c Sac7d Sso7d AffitinNotes
Acknowledgments
The authors thank all previous members of the laboratory who helped to develop this protocol. In memoriam of Ghislaine Béhar.
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