Panning and selection of proteins using ribosome display

Eukaryotic ribosome complexes can be used as a means to display a library o f proteins, and isolate specific binding reagents by screening against targ et molecules. Here we present, as an example, a method for the display of a library of immunoglobulin variable-like domains (VLDs) for the production of stable mRNA/ribosome/protein complexes. These complexes are produced by the addition of specific in vitro transcriptional promoter elements and tra nslation control sequences to the template DNA. Furthermore, an appropriate spacer (anchor) domain is included for efficient folding of the nascent tr anslated protein. which remains attached to the ribosome complex. Ribosome complexes are panned against hen egg lysozyme-conjugated magnetic beads and genes encoding specific, binding, V-like domains are recovered by RT-PCR a nd cloned into an Escherichia coli expression vector. (C) 2001 Elsevier Sci ence B.V. All rights reserved.