In vitro selection of other proteins than antibodies by means of ribosome display

The in vitro selection and simultaneous evolution of proteins is feasible b y means of ribosome display. Here, we describe the use of a protein bearing a binding property without being an antibody for affinity enrichment of th e ternary complex, consisting of a protein, a ribosome and a encoding mRNA. The binding property was a simple affinity tag, namely Strep-tag II and Hi s-tag. We could demonstrate that the selection of a specific mRNA encoding a shortened bovine heart fatty acid binding protein with a N-terminal His-t ag was possible. After nine cycles of transcription, translation, affinity selection and reverse transcription PCR the protein with the His-tag could be enriched 10(8)-fold. (C) 2001 Federation of European Biochemical Societi es. Published by Elsevier Science B.V. All rights reserved.