Affitins: Ribosome Display for Selection of Aho7c-Based Affinity Proteins
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 wordsRibosome display In vitro selection Sul7d Aho7c Sac7d Sso7d Affitin
The authors thank all previous members of the laboratory who helped to develop this protocol. In memoriam of Ghislaine Béhar.
- 2.Pecorari F, Alzari PM (2008) OB-fold used as scaffold for engineering new specific binders. Patent Publication Nos PCT/IB2007/004388Google Scholar
- 14.Vukojicic P, Béhar G, Tawara MH, Fernandez-Villamarin M, Pecorari F, Fernandez-Megia E, Mouratou B (2019) Multivalent Affidendrons with High Affinity and Specificity toward as Versatile Tools for Modulating Multicellular Behaviors. ACS Appl Mater Interfaces 11(24):21391–21398Google Scholar
- 22.Amstutz P, Binz HK, Zahnd C, Pluckthun A (2006) Ribosome display: in vitro selection of protein-protein interactions. In: Celis J (ed) Cell biology—a laboratory handbook, vol 1. Elsevier Academic Press, Cambridge, pp 497–509Google Scholar