Figure 4.

Schematic representation of a ribosome display selection cycle. An mRNA library encoding the proteins of interest without stop codon is translated in vitro (1). After cooling, the translation yields stable ternary complexes of mRNA, ribosomes and nascent polypeptides. These complexes are used for the binding selection on the immobilized target (2). After binding of the polypeptides to the target protein, unbound complexes are washed away (3). The mRNA of the bound complex is eluted by dissociating the ribosomal complex with EDTA (4). A reverse transcription reaction followed by PCR yields the genetic information of the selected clones (5). The amplified genes can then used as input for the next selection round starting with in vitro transcription (6) or cloned intro plasmids for analysis. (adapted from [56]).